                         SEQUENCE LISTING

<110>  Council of Scientific & Industrial Research
       Garg, Roma
       Sahni, Girish
 
<120>  Cellulase derived from metagenomics

<130>  -

<160>  45    

<170>  PatentIn version 3.5

<210>  1
<211>  1017
<212>  DNA
<213>  unknown

<220>
<223>  Soil metagenome derived nucleotide sequence

<400>  1
atgaagaaaa actcaatcat tctcacactg gttctttttg ttattgttgc tttgtcctgt       60

accggcagca gtaaaaaaac ggaaaacaac aggaaaacgg actaccgttc catcgtagcg      120

cagaacggcc gtttacaggt gatcggcaca caattgagca atgaaaaagg cgaacccgtc      180

gttttacggg gagccagcct cggatggcac aacctctggc cccgcttcta taacaagaac      240

gcggtgcaat ggctggccga cgattggaaa tgcacggtcg tcagggctgc aatgggcttg      300

gaaattgaag acaactaccg ggaaaatccc gaatttgcct tacagtgcat cacccctgtc      360

atcgaatcgg ctattgaaaa cggaatctat gtgattatcg acttccacgc acacaacaaa      420

tacaccgaag aagccaaaac attctttgcc gggatggccg aaaaatacgg ggaatatccg      480

aacgtgatct atgaaatctg gaacgaaccc gattatttcg aatgggaaga agtaaaaacc      540

tattcggaag aagtgatcgc cgtcatccgc gcgatcgacc ccgacaatat tatattggtc      600

ggcagccccc attgggacca ggacctgcat ctggtagccg aagacccgat ccgggatgta      660

agcaacatca tgtacaccat gcatttttac gccgcgaccc acgaggcctg gctgcgtgac      720

cggaccgacg aggcgattgc caaaggaatt cccgttttcg tatcggagtg cggcggctcg      780

gaagccaatg gcgacggacg gttaggcata gaagaatgga aaacctacgt cgattggatg      840

gagagtcgga aaataagctg ggtggcctgg tccgtttccg acaagaacga aacctgctcc      900

atgctgctcc cccgcgcctc tgccgatggc aactggacgg aagacctgct caagccttgg      960

ggaaaactga cgcgtaattc tatccggaac gcgaatgatg aaaacccgga tatctga        1017


<210>  2
<211>  338
<212>  PRT
<213>  unknown

<220>
<223>  Polypeptide sequence of the cellulase derived from soil 
       metagenome

<400>  2

Met Lys Lys Asn Ser Ile Ile Leu Thr Leu Val Leu Phe Val Ile Val 
1               5                   10                  15      


Ala Leu Ser Cys Thr Gly Ser Ser Lys Lys Thr Glu Asn Asn Arg Lys 
            20                  25                  30          


Thr Asp Tyr Arg Ser Ile Val Ala Gln Asn Gly Arg Leu Gln Val Ile 
        35                  40                  45              


Gly Thr Gln Leu Ser Asn Glu Lys Gly Glu Pro Val Val Leu Arg Gly 
    50                  55                  60                  


Ala Ser Leu Gly Trp His Asn Leu Trp Pro Arg Phe Tyr Asn Lys Asn 
65                  70                  75                  80  


Ala Val Gln Trp Leu Ala Asp Asp Trp Lys Cys Thr Val Val Arg Ala 
                85                  90                  95      


Ala Met Gly Leu Glu Ile Glu Asp Asn Tyr Arg Glu Asn Pro Glu Phe 
            100                 105                 110         


Ala Leu Gln Cys Ile Thr Pro Val Ile Glu Ser Ala Ile Glu Asn Gly 
        115                 120                 125             


Ile Tyr Val Ile Ile Asp Phe His Ala His Asn Lys Tyr Thr Glu Glu 
    130                 135                 140                 


Ala Lys Thr Phe Phe Ala Gly Met Ala Glu Lys Tyr Gly Glu Tyr Pro 
145                 150                 155                 160 


Asn Val Ile Tyr Glu Ile Trp Asn Glu Pro Asp Tyr Phe Glu Trp Glu 
                165                 170                 175     


Glu Val Lys Thr Tyr Ser Glu Glu Val Ile Ala Val Ile Arg Ala Ile 
            180                 185                 190         


Asp Pro Asp Asn Ile Ile Leu Val Gly Ser Pro His Trp Asp Gln Asp 
        195                 200                 205             


Leu His Leu Val Ala Glu Asp Pro Ile Arg Asp Val Ser Asn Ile Met 
    210                 215                 220                 


Tyr Thr Met His Phe Tyr Ala Ala Thr His Glu Ala Trp Leu Arg Asp 
225                 230                 235                 240 


Arg Thr Asp Glu Ala Ile Ala Lys Gly Ile Pro Val Phe Val Ser Glu 
                245                 250                 255     


Cys Gly Gly Ser Glu Ala Asn Gly Asp Gly Arg Leu Gly Ile Glu Glu 
            260                 265                 270         


Trp Lys Thr Tyr Val Asp Trp Met Glu Ser Arg Lys Ile Ser Trp Val 
        275                 280                 285             


Ala Trp Ser Val Ser Asp Lys Asn Glu Thr Cys Ser Met Leu Leu Pro 
    290                 295                 300                 


Arg Ala Ser Ala Asp Gly Asn Trp Thr Glu Asp Leu Leu Lys Pro Trp 
305                 310                 315                 320 


Gly Lys Leu Thr Arg Asn Ser Ile Arg Asn Ala Asn Asp Glu Asn Pro 
                325                 330                 335     


Asp Ile 
        


<210>  3
<211>  936
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Sequence derived from soil metagenome, particularly nucleotide 
       sequence of the gene after truncating the upstream hydrophobic 
       patch

<400>  3
gaaaacaaca ggaaaacgga ctaccgttcc atcgtagcgc agaacggccg tttacaggtg       60

atcggcacac aattgagcaa tgaaaaaggc gaacccgtcg ttttacgggg agccagcctc      120

ggatggcaca acctctggcc ccgcttctat aacaagaacg cggtgcaatg gctggccgac      180

gattggaaat gcacggtcgt cagggctgca atgggcttgg aaattgaaga caactaccgg      240

gaaaatcccg aatttgcctt acagtgcatc acccctgtca tcgaatcggc tattgaaaac      300

ggaatctatg tgattatcga cttccacgca cacaacaaat acaccgaaga agccaaaaca      360

ttctttgccg ggatggccga aaaatacggg gaatatccga acgtgatcta tgaaatctgg      420

aacgaacccg attatttcga atgggaagaa gtaaaaacct attcggaaga agtgatcgcc      480

gtcatccgcg cgatcgaccc cgacaatatt atattggtcg gcagccccca ttgggaccag      540

gacctgcatc tggtagccga agacccgatc cgggatgtaa gcaacatcat gtacaccatg      600

catttttacg ccgcgaccca cgaggcctgg ctgcgtgacc ggaccgacga ggcgattgcc      660

aaaggaattc ccgttttcgt atcggagtgc ggcggctcgg aagccaatgg cgacggacgg      720

ttaggcatag aagaatggaa aacctacgtc gattggatgg agagtcggaa aataagctgg      780

gtggcctggt ccgtttccga caagaacgaa acctgctcca tgctgctccc ccgcgcctct      840

gccgatggca actggacgga agacctgctc aagccttggg gaaaactgac gcgtaattct      900

atccggaacg cgaatgatga aaacccggat atctga                                936


<210>  4
<211>  311
<212>  PRT
<213>  Artificial sequence

<220>
<223>  Sequence derived from soil metagenome, particularly polypeptide 
       sequence of the gene after truncating the upstream hydrophobic 
       patch

<400>  4

Glu Asn Asn Arg Lys Thr Asp Tyr Arg Ser Ile Val Ala Gln Asn Gly 
1               5                   10                  15      


Arg Leu Gln Val Ile Gly Thr Gln Leu Ser Asn Glu Lys Gly Glu Pro 
            20                  25                  30          


Val Val Leu Arg Gly Ala Ser Leu Gly Trp His Asn Leu Trp Pro Arg 
        35                  40                  45              


Phe Tyr Asn Lys Asn Ala Val Gln Trp Leu Ala Asp Asp Trp Lys Cys 
    50                  55                  60                  


Thr Val Val Arg Ala Ala Met Gly Leu Glu Ile Glu Asp Asn Tyr Arg 
65                  70                  75                  80  


Glu Asn Pro Glu Phe Ala Leu Gln Cys Ile Thr Pro Val Ile Glu Ser 
                85                  90                  95      


Ala Ile Glu Asn Gly Ile Tyr Val Ile Ile Asp Phe His Ala His Asn 
            100                 105                 110         


Lys Tyr Thr Glu Glu Ala Lys Thr Phe Phe Ala Gly Met Ala Glu Lys 
        115                 120                 125             


Tyr Gly Glu Tyr Pro Asn Val Ile Tyr Glu Ile Trp Asn Glu Pro Asp 
    130                 135                 140                 


Tyr Phe Glu Trp Glu Glu Val Lys Thr Tyr Ser Glu Glu Val Ile Ala 
145                 150                 155                 160 


Val Ile Arg Ala Ile Asp Pro Asp Asn Ile Ile Leu Val Gly Ser Pro 
                165                 170                 175     


His Trp Asp Gln Asp Leu His Leu Val Ala Glu Asp Pro Ile Arg Asp 
            180                 185                 190         


Val Ser Asn Ile Met Tyr Thr Met His Phe Tyr Ala Ala Thr His Glu 
        195                 200                 205             


Ala Trp Leu Arg Asp Arg Thr Asp Glu Ala Ile Ala Lys Gly Ile Pro 
    210                 215                 220                 


Val Phe Val Ser Glu Cys Gly Gly Ser Glu Ala Asn Gly Asp Gly Arg 
225                 230                 235                 240 


Leu Gly Ile Glu Glu Trp Lys Thr Tyr Val Asp Trp Met Glu Ser Arg 
                245                 250                 255     


Lys Ile Ser Trp Val Ala Trp Ser Val Ser Asp Lys Asn Glu Thr Cys 
            260                 265                 270         


Ser Met Leu Leu Pro Arg Ala Ser Ala Asp Gly Asn Trp Thr Glu Asp 
        275                 280                 285             


Leu Leu Lys Pro Trp Gly Lys Leu Thr Arg Asn Ser Ile Arg Asn Ala 
    290                 295                 300                 


Asn Asp Glu Asn Pro Asp Ile 
305                 310     


<210>  5
<211>  311
<212>  PRT
<213>  Artificial sequence

<220>
<223>  Sequence representing single alanine substitution made on SEQ ID 
       NO 4 wherein cysteine 64 is replaced by alanine

<400>  5

Glu Asn Asn Arg Lys Thr Asp Tyr Arg Ser Ile Val Ala Gln Asn Gly 
1               5                   10                  15      


Arg Leu Gln Val Ile Gly Thr Gln Leu Ser Asn Glu Lys Gly Glu Pro 
            20                  25                  30          


Val Val Leu Arg Gly Ala Ser Leu Gly Trp His Asn Leu Trp Pro Arg 
        35                  40                  45              


Phe Tyr Asn Lys Asn Ala Val Gln Trp Leu Ala Asp Asp Trp Lys Ala 
    50                  55                  60                  


Thr Val Val Arg Ala Ala Met Gly Leu Glu Ile Glu Asp Asn Tyr Arg 
65                  70                  75                  80  


Glu Asn Pro Glu Phe Ala Leu Gln Cys Ile Thr Pro Val Ile Glu Ser 
                85                  90                  95      


Ala Ile Glu Asn Gly Ile Tyr Val Ile Ile Asp Phe His Ala His Asn 
            100                 105                 110         


Lys Tyr Thr Glu Glu Ala Lys Thr Phe Phe Ala Gly Met Ala Glu Lys 
        115                 120                 125             


Tyr Gly Glu Tyr Pro Asn Val Ile Tyr Glu Ile Trp Asn Glu Pro Asp 
    130                 135                 140                 


Tyr Phe Glu Trp Glu Glu Val Lys Thr Tyr Ser Glu Glu Val Ile Ala 
145                 150                 155                 160 


Val Ile Arg Ala Ile Asp Pro Asp Asn Ile Ile Leu Val Gly Ser Pro 
                165                 170                 175     


His Trp Asp Gln Asp Leu His Leu Val Ala Glu Asp Pro Ile Arg Asp 
            180                 185                 190         


Val Ser Asn Ile Met Tyr Thr Met His Phe Tyr Ala Ala Thr His Glu 
        195                 200                 205             


Ala Trp Leu Arg Asp Arg Thr Asp Glu Ala Ile Ala Lys Gly Ile Pro 
    210                 215                 220                 


Val Phe Val Ser Glu Cys Gly Gly Ser Glu Ala Asn Gly Asp Gly Arg 
225                 230                 235                 240 


Leu Gly Ile Glu Glu Trp Lys Thr Tyr Val Asp Trp Met Glu Ser Arg 
                245                 250                 255     


Lys Ile Ser Trp Val Ala Trp Ser Val Ser Asp Lys Asn Glu Thr Cys 
            260                 265                 270         


Ser Met Leu Leu Pro Arg Ala Ser Ala Asp Gly Asn Trp Thr Glu Asp 
        275                 280                 285             


Leu Leu Lys Pro Trp Gly Lys Leu Thr Arg Asn Ser Ile Arg Asn Ala 
    290                 295                 300                 


Asn Asp Glu Asn Pro Asp Ile 
305                 310     


<210>  6
<211>  311
<212>  PRT
<213>  Artificial sequence

<220>
<223>  Sequence representing single alanine substitution made on SEQ ID 
       NO 4 wherein cysteine 89 is replaced by alanine

<400>  6

Glu Asn Asn Arg Lys Thr Asp Tyr Arg Ser Ile Val Ala Gln Asn Gly 
1               5                   10                  15      


Arg Leu Gln Val Ile Gly Thr Gln Leu Ser Asn Glu Lys Gly Glu Pro 
            20                  25                  30          


Val Val Leu Arg Gly Ala Ser Leu Gly Trp His Asn Leu Trp Pro Arg 
        35                  40                  45              


Phe Tyr Asn Lys Asn Ala Val Gln Trp Leu Ala Asp Asp Trp Lys Cys 
    50                  55                  60                  


Thr Val Val Arg Ala Ala Met Gly Leu Glu Ile Glu Asp Asn Tyr Arg 
65                  70                  75                  80  


Glu Asn Pro Glu Phe Ala Leu Gln Ala Ile Thr Pro Val Ile Glu Ser 
                85                  90                  95      


Ala Ile Glu Asn Gly Ile Tyr Val Ile Ile Asp Phe His Ala His Asn 
            100                 105                 110         


Lys Tyr Thr Glu Glu Ala Lys Thr Phe Phe Ala Gly Met Ala Glu Lys 
        115                 120                 125             


Tyr Gly Glu Tyr Pro Asn Val Ile Tyr Glu Ile Trp Asn Glu Pro Asp 
    130                 135                 140                 


Tyr Phe Glu Trp Glu Glu Val Lys Thr Tyr Ser Glu Glu Val Ile Ala 
145                 150                 155                 160 


Val Ile Arg Ala Ile Asp Pro Asp Asn Ile Ile Leu Val Gly Ser Pro 
                165                 170                 175     


His Trp Asp Gln Asp Leu His Leu Val Ala Glu Asp Pro Ile Arg Asp 
            180                 185                 190         


Val Ser Asn Ile Met Tyr Thr Met His Phe Tyr Ala Ala Thr His Glu 
        195                 200                 205             


Ala Trp Leu Arg Asp Arg Thr Asp Glu Ala Ile Ala Lys Gly Ile Pro 
    210                 215                 220                 


Val Phe Val Ser Glu Cys Gly Gly Ser Glu Ala Asn Gly Asp Gly Arg 
225                 230                 235                 240 


Leu Gly Ile Glu Glu Trp Lys Thr Tyr Val Asp Trp Met Glu Ser Arg 
                245                 250                 255     


Lys Ile Ser Trp Val Ala Trp Ser Val Ser Asp Lys Asn Glu Thr Cys 
            260                 265                 270         


Ser Met Leu Leu Pro Arg Ala Ser Ala Asp Gly Asn Trp Thr Glu Asp 
        275                 280                 285             


Leu Leu Lys Pro Trp Gly Lys Leu Thr Arg Asn Ser Ile Arg Asn Ala 
    290                 295                 300                 


Asn Asp Glu Asn Pro Asp Ile 
305                 310     


<210>  7
<211>  311
<212>  PRT
<213>  Artificial sequence

<220>
<223>  Sequence representing single alanine substitution made on SEQ ID 
       NO 4 wherein cysteine 230 is replaced by alanine

<400>  7

Glu Asn Asn Arg Lys Thr Asp Tyr Arg Ser Ile Val Ala Gln Asn Gly 
1               5                   10                  15      


Arg Leu Gln Val Ile Gly Thr Gln Leu Ser Asn Glu Lys Gly Glu Pro 
            20                  25                  30          


Val Val Leu Arg Gly Ala Ser Leu Gly Trp His Asn Leu Trp Pro Arg 
        35                  40                  45              


Phe Tyr Asn Lys Asn Ala Val Gln Trp Leu Ala Asp Asp Trp Lys Cys 
    50                  55                  60                  


Thr Val Val Arg Ala Ala Met Gly Leu Glu Ile Glu Asp Asn Tyr Arg 
65                  70                  75                  80  


Glu Asn Pro Glu Phe Ala Leu Gln Cys Ile Thr Pro Val Ile Glu Ser 
                85                  90                  95      


Ala Ile Glu Asn Gly Ile Tyr Val Ile Ile Asp Phe His Ala His Asn 
            100                 105                 110         


Lys Tyr Thr Glu Glu Ala Lys Thr Phe Phe Ala Gly Met Ala Glu Lys 
        115                 120                 125             


Tyr Gly Glu Tyr Pro Asn Val Ile Tyr Glu Ile Trp Asn Glu Pro Asp 
    130                 135                 140                 


Tyr Phe Glu Trp Glu Glu Val Lys Thr Tyr Ser Glu Glu Val Ile Ala 
145                 150                 155                 160 


Val Ile Arg Ala Ile Asp Pro Asp Asn Ile Ile Leu Val Gly Ser Pro 
                165                 170                 175     


His Trp Asp Gln Asp Leu His Leu Val Ala Glu Asp Pro Ile Arg Asp 
            180                 185                 190         


Val Ser Asn Ile Met Tyr Thr Met His Phe Tyr Ala Ala Thr His Glu 
        195                 200                 205             


Ala Trp Leu Arg Asp Arg Thr Asp Glu Ala Ile Ala Lys Gly Ile Pro 
    210                 215                 220                 


Val Phe Val Ser Glu Ala Gly Gly Ser Glu Ala Asn Gly Asp Gly Arg 
225                 230                 235                 240 


Leu Gly Ile Glu Glu Trp Lys Thr Tyr Val Asp Trp Met Glu Ser Arg 
                245                 250                 255     


Lys Ile Ser Trp Val Ala Trp Ser Val Ser Asp Lys Asn Glu Thr Cys 
            260                 265                 270         


Ser Met Leu Leu Pro Arg Ala Ser Ala Asp Gly Asn Trp Thr Glu Asp 
        275                 280                 285             


Leu Leu Lys Pro Trp Gly Lys Leu Thr Arg Asn Ser Ile Arg Asn Ala 
    290                 295                 300                 


Asn Asp Glu Asn Pro Asp Ile 
305                 310     


<210>  8
<211>  311
<212>  PRT
<213>  Artificial sequence

<220>
<223>  Sequence representing single alanine substitution made on SEQ ID 
       NO 4 wherein cysteine 272 is replaced by alanine

<400>  8

Glu Asn Asn Arg Lys Thr Asp Tyr Arg Ser Ile Val Ala Gln Asn Gly 
1               5                   10                  15      


Arg Leu Gln Val Ile Gly Thr Gln Leu Ser Asn Glu Lys Gly Glu Pro 
            20                  25                  30          


Val Val Leu Arg Gly Ala Ser Leu Gly Trp His Asn Leu Trp Pro Arg 
        35                  40                  45              


Phe Tyr Asn Lys Asn Ala Val Gln Trp Leu Ala Asp Asp Trp Lys Cys 
    50                  55                  60                  


Thr Val Val Arg Ala Ala Met Gly Leu Glu Ile Glu Asp Asn Tyr Arg 
65                  70                  75                  80  


Glu Asn Pro Glu Phe Ala Leu Gln Cys Ile Thr Pro Val Ile Glu Ser 
                85                  90                  95      


Ala Ile Glu Asn Gly Ile Tyr Val Ile Ile Asp Phe His Ala His Asn 
            100                 105                 110         


Lys Tyr Thr Glu Glu Ala Lys Thr Phe Phe Ala Gly Met Ala Glu Lys 
        115                 120                 125             


Tyr Gly Glu Tyr Pro Asn Val Ile Tyr Glu Ile Trp Asn Glu Pro Asp 
    130                 135                 140                 


Tyr Phe Glu Trp Glu Glu Val Lys Thr Tyr Ser Glu Glu Val Ile Ala 
145                 150                 155                 160 


Val Ile Arg Ala Ile Asp Pro Asp Asn Ile Ile Leu Val Gly Ser Pro 
                165                 170                 175     


His Trp Asp Gln Asp Leu His Leu Val Ala Glu Asp Pro Ile Arg Asp 
            180                 185                 190         


Val Ser Asn Ile Met Tyr Thr Met His Phe Tyr Ala Ala Thr His Glu 
        195                 200                 205             


Ala Trp Leu Arg Asp Arg Thr Asp Glu Ala Ile Ala Lys Gly Ile Pro 
    210                 215                 220                 


Val Phe Val Ser Glu Cys Gly Gly Ser Glu Ala Asn Gly Asp Gly Arg 
225                 230                 235                 240 


Leu Gly Ile Glu Glu Trp Lys Thr Tyr Val Asp Trp Met Glu Ser Arg 
                245                 250                 255     


Lys Ile Ser Trp Val Ala Trp Ser Val Ser Asp Lys Asn Glu Thr Ala 
            260                 265                 270         


Ser Met Leu Leu Pro Arg Ala Ser Ala Asp Gly Asn Trp Thr Glu Asp 
        275                 280                 285             


Leu Leu Lys Pro Trp Gly Lys Leu Thr Arg Asn Ser Ile Arg Asn Ala 
    290                 295                 300                 


Asn Asp Glu Asn Pro Asp Ile 
305                 310     


<210>  9
<211>  311
<212>  PRT
<213>  Artificial sequence

<220>
<223>  Sequence representing double alanine substitutions made on SEQ ID
       NO 4 wherein cysteine 64 and 89 are replaced by alanine

<400>  9

Glu Asn Asn Arg Lys Thr Asp Tyr Arg Ser Ile Val Ala Gln Asn Gly 
1               5                   10                  15      


Arg Leu Gln Val Ile Gly Thr Gln Leu Ser Asn Glu Lys Gly Glu Pro 
            20                  25                  30          


Val Val Leu Arg Gly Ala Ser Leu Gly Trp His Asn Leu Trp Pro Arg 
        35                  40                  45              


Phe Tyr Asn Lys Asn Ala Val Gln Trp Leu Ala Asp Asp Trp Lys Ala 
    50                  55                  60                  


Thr Val Val Arg Ala Ala Met Gly Leu Glu Ile Glu Asp Asn Tyr Arg 
65                  70                  75                  80  


Glu Asn Pro Glu Phe Ala Leu Gln Ala Ile Thr Pro Val Ile Glu Ser 
                85                  90                  95      


Ala Ile Glu Asn Gly Ile Tyr Val Ile Ile Asp Phe His Ala His Asn 
            100                 105                 110         


Lys Tyr Thr Glu Glu Ala Lys Thr Phe Phe Ala Gly Met Ala Glu Lys 
        115                 120                 125             


Tyr Gly Glu Tyr Pro Asn Val Ile Tyr Glu Ile Trp Asn Glu Pro Asp 
    130                 135                 140                 


Tyr Phe Glu Trp Glu Glu Val Lys Thr Tyr Ser Glu Glu Val Ile Ala 
145                 150                 155                 160 


Val Ile Arg Ala Ile Asp Pro Asp Asn Ile Ile Leu Val Gly Ser Pro 
                165                 170                 175     


His Trp Asp Gln Asp Leu His Leu Val Ala Glu Asp Pro Ile Arg Asp 
            180                 185                 190         


Val Ser Asn Ile Met Tyr Thr Met His Phe Tyr Ala Ala Thr His Glu 
        195                 200                 205             


Ala Trp Leu Arg Asp Arg Thr Asp Glu Ala Ile Ala Lys Gly Ile Pro 
    210                 215                 220                 


Val Phe Val Ser Glu Cys Gly Gly Ser Glu Ala Asn Gly Asp Gly Arg 
225                 230                 235                 240 


Leu Gly Ile Glu Glu Trp Lys Thr Tyr Val Asp Trp Met Glu Ser Arg 
                245                 250                 255     


Lys Ile Ser Trp Val Ala Trp Ser Val Ser Asp Lys Asn Glu Thr Cys 
            260                 265                 270         


Ser Met Leu Leu Pro Arg Ala Ser Ala Asp Gly Asn Trp Thr Glu Asp 
        275                 280                 285             


Leu Leu Lys Pro Trp Gly Lys Leu Thr Arg Asn Ser Ile Arg Asn Ala 
    290                 295                 300                 


Asn Asp Glu Asn Pro Asp Ile 
305                 310     


<210>  10
<211>  311
<212>  PRT
<213>  Artificial sequence

<220>
<223>  Sequence representing double alanine substitutions made on SEQ ID
       NO 4 wherein cysteine 64 and 230 are replaced by alanine

<400>  10

Glu Asn Asn Arg Lys Thr Asp Tyr Arg Ser Ile Val Ala Gln Asn Gly 
1               5                   10                  15      


Arg Leu Gln Val Ile Gly Thr Gln Leu Ser Asn Glu Lys Gly Glu Pro 
            20                  25                  30          


Val Val Leu Arg Gly Ala Ser Leu Gly Trp His Asn Leu Trp Pro Arg 
        35                  40                  45              


Phe Tyr Asn Lys Asn Ala Val Gln Trp Leu Ala Asp Asp Trp Lys Ala 
    50                  55                  60                  


Thr Val Val Arg Ala Ala Met Gly Leu Glu Ile Glu Asp Asn Tyr Arg 
65                  70                  75                  80  


Glu Asn Pro Glu Phe Ala Leu Gln Cys Ile Thr Pro Val Ile Glu Ser 
                85                  90                  95      


Ala Ile Glu Asn Gly Ile Tyr Val Ile Ile Asp Phe His Ala His Asn 
            100                 105                 110         


Lys Tyr Thr Glu Glu Ala Lys Thr Phe Phe Ala Gly Met Ala Glu Lys 
        115                 120                 125             


Tyr Gly Glu Tyr Pro Asn Val Ile Tyr Glu Ile Trp Asn Glu Pro Asp 
    130                 135                 140                 


Tyr Phe Glu Trp Glu Glu Val Lys Thr Tyr Ser Glu Glu Val Ile Ala 
145                 150                 155                 160 


Val Ile Arg Ala Ile Asp Pro Asp Asn Ile Ile Leu Val Gly Ser Pro 
                165                 170                 175     


His Trp Asp Gln Asp Leu His Leu Val Ala Glu Asp Pro Ile Arg Asp 
            180                 185                 190         


Val Ser Asn Ile Met Tyr Thr Met His Phe Tyr Ala Ala Thr His Glu 
        195                 200                 205             


Ala Trp Leu Arg Asp Arg Thr Asp Glu Ala Ile Ala Lys Gly Ile Pro 
    210                 215                 220                 


Val Phe Val Ser Glu Ala Gly Gly Ser Glu Ala Asn Gly Asp Gly Arg 
225                 230                 235                 240 


Leu Gly Ile Glu Glu Trp Lys Thr Tyr Val Asp Trp Met Glu Ser Arg 
                245                 250                 255     


Lys Ile Ser Trp Val Ala Trp Ser Val Ser Asp Lys Asn Glu Thr Cys 
            260                 265                 270         


Ser Met Leu Leu Pro Arg Ala Ser Ala Asp Gly Asn Trp Thr Glu Asp 
        275                 280                 285             


Leu Leu Lys Pro Trp Gly Lys Leu Thr Arg Asn Ser Ile Arg Asn Ala 
    290                 295                 300                 


Asn Asp Glu Asn Pro Asp Ile 
305                 310     


<210>  11
<211>  311
<212>  PRT
<213>  Artificial sequence

<220>
<223>  Sequence representing double alanine substitutions made on SEQ ID
       NO 4 wherein cysteine 64 and 272 are replaced by alanine

<400>  11

Glu Asn Asn Arg Lys Thr Asp Tyr Arg Ser Ile Val Ala Gln Asn Gly 
1               5                   10                  15      


Arg Leu Gln Val Ile Gly Thr Gln Leu Ser Asn Glu Lys Gly Glu Pro 
            20                  25                  30          


Val Val Leu Arg Gly Ala Ser Leu Gly Trp His Asn Leu Trp Pro Arg 
        35                  40                  45              


Phe Tyr Asn Lys Asn Ala Val Gln Trp Leu Ala Asp Asp Trp Lys Ala 
    50                  55                  60                  


Thr Val Val Arg Ala Ala Met Gly Leu Glu Ile Glu Asp Asn Tyr Arg 
65                  70                  75                  80  


Glu Asn Pro Glu Phe Ala Leu Gln Cys Ile Thr Pro Val Ile Glu Ser 
                85                  90                  95      


Ala Ile Glu Asn Gly Ile Tyr Val Ile Ile Asp Phe His Ala His Asn 
            100                 105                 110         


Lys Tyr Thr Glu Glu Ala Lys Thr Phe Phe Ala Gly Met Ala Glu Lys 
        115                 120                 125             


Tyr Gly Glu Tyr Pro Asn Val Ile Tyr Glu Ile Trp Asn Glu Pro Asp 
    130                 135                 140                 


Tyr Phe Glu Trp Glu Glu Val Lys Thr Tyr Ser Glu Glu Val Ile Ala 
145                 150                 155                 160 


Val Ile Arg Ala Ile Asp Pro Asp Asn Ile Ile Leu Val Gly Ser Pro 
                165                 170                 175     


His Trp Asp Gln Asp Leu His Leu Val Ala Glu Asp Pro Ile Arg Asp 
            180                 185                 190         


Val Ser Asn Ile Met Tyr Thr Met His Phe Tyr Ala Ala Thr His Glu 
        195                 200                 205             


Ala Trp Leu Arg Asp Arg Thr Asp Glu Ala Ile Ala Lys Gly Ile Pro 
    210                 215                 220                 


Val Phe Val Ser Glu Cys Gly Gly Ser Glu Ala Asn Gly Asp Gly Arg 
225                 230                 235                 240 


Leu Gly Ile Glu Glu Trp Lys Thr Tyr Val Asp Trp Met Glu Ser Arg 
                245                 250                 255     


Lys Ile Ser Trp Val Ala Trp Ser Val Ser Asp Lys Asn Glu Thr Ala 
            260                 265                 270         


Ser Met Leu Leu Pro Arg Ala Ser Ala Asp Gly Asn Trp Thr Glu Asp 
        275                 280                 285             


Leu Leu Lys Pro Trp Gly Lys Leu Thr Arg Asn Ser Ile Arg Asn Ala 
    290                 295                 300                 


Asn Asp Glu Asn Pro Asp Ile 
305                 310     


<210>  12
<211>  311
<212>  PRT
<213>  Artificial sequence

<220>
<223>  Sequence representing double alanine substitutions made on SEQ ID
       NO 4 wherein cysteine 89 and 230 are replaced by alanine

<400>  12

Glu Asn Asn Arg Lys Thr Asp Tyr Arg Ser Ile Val Ala Gln Asn Gly 
1               5                   10                  15      


Arg Leu Gln Val Ile Gly Thr Gln Leu Ser Asn Glu Lys Gly Glu Pro 
            20                  25                  30          


Val Val Leu Arg Gly Ala Ser Leu Gly Trp His Asn Leu Trp Pro Arg 
        35                  40                  45              


Phe Tyr Asn Lys Asn Ala Val Gln Trp Leu Ala Asp Asp Trp Lys Cys 
    50                  55                  60                  


Thr Val Val Arg Ala Ala Met Gly Leu Glu Ile Glu Asp Asn Tyr Arg 
65                  70                  75                  80  


Glu Asn Pro Glu Phe Ala Leu Gln Ala Ile Thr Pro Val Ile Glu Ser 
                85                  90                  95      


Ala Ile Glu Asn Gly Ile Tyr Val Ile Ile Asp Phe His Ala His Asn 
            100                 105                 110         


Lys Tyr Thr Glu Glu Ala Lys Thr Phe Phe Ala Gly Met Ala Glu Lys 
        115                 120                 125             


Tyr Gly Glu Tyr Pro Asn Val Ile Tyr Glu Ile Trp Asn Glu Pro Asp 
    130                 135                 140                 


Tyr Phe Glu Trp Glu Glu Val Lys Thr Tyr Ser Glu Glu Val Ile Ala 
145                 150                 155                 160 


Val Ile Arg Ala Ile Asp Pro Asp Asn Ile Ile Leu Val Gly Ser Pro 
                165                 170                 175     


His Trp Asp Gln Asp Leu His Leu Val Ala Glu Asp Pro Ile Arg Asp 
            180                 185                 190         


Val Ser Asn Ile Met Tyr Thr Met His Phe Tyr Ala Ala Thr His Glu 
        195                 200                 205             


Ala Trp Leu Arg Asp Arg Thr Asp Glu Ala Ile Ala Lys Gly Ile Pro 
    210                 215                 220                 


Val Phe Val Ser Glu Ala Gly Gly Ser Glu Ala Asn Gly Asp Gly Arg 
225                 230                 235                 240 


Leu Gly Ile Glu Glu Trp Lys Thr Tyr Val Asp Trp Met Glu Ser Arg 
                245                 250                 255     


Lys Ile Ser Trp Val Ala Trp Ser Val Ser Asp Lys Asn Glu Thr Cys 
            260                 265                 270         


Ser Met Leu Leu Pro Arg Ala Ser Ala Asp Gly Asn Trp Thr Glu Asp 
        275                 280                 285             


Leu Leu Lys Pro Trp Gly Lys Leu Thr Arg Asn Ser Ile Arg Asn Ala 
    290                 295                 300                 


Asn Asp Glu Asn Pro Asp Ile 
305                 310     


<210>  13
<211>  311
<212>  PRT
<213>  Artificial sequence

<220>
<223>  Sequence representing double alanine substitutions made on SEQ ID
       NO 4 wherein cysteine 89 and 272 are replaced by alanine

<400>  13

Glu Asn Asn Arg Lys Thr Asp Tyr Arg Ser Ile Val Ala Gln Asn Gly 
1               5                   10                  15      


Arg Leu Gln Val Ile Gly Thr Gln Leu Ser Asn Glu Lys Gly Glu Pro 
            20                  25                  30          


Val Val Leu Arg Gly Ala Ser Leu Gly Trp His Asn Leu Trp Pro Arg 
        35                  40                  45              


Phe Tyr Asn Lys Asn Ala Val Gln Trp Leu Ala Asp Asp Trp Lys Cys 
    50                  55                  60                  


Thr Val Val Arg Ala Ala Met Gly Leu Glu Ile Glu Asp Asn Tyr Arg 
65                  70                  75                  80  


Glu Asn Pro Glu Phe Ala Leu Gln Ala Ile Thr Pro Val Ile Glu Ser 
                85                  90                  95      


Ala Ile Glu Asn Gly Ile Tyr Val Ile Ile Asp Phe His Ala His Asn 
            100                 105                 110         


Lys Tyr Thr Glu Glu Ala Lys Thr Phe Phe Ala Gly Met Ala Glu Lys 
        115                 120                 125             


Tyr Gly Glu Tyr Pro Asn Val Ile Tyr Glu Ile Trp Asn Glu Pro Asp 
    130                 135                 140                 


Tyr Phe Glu Trp Glu Glu Val Lys Thr Tyr Ser Glu Glu Val Ile Ala 
145                 150                 155                 160 


Val Ile Arg Ala Ile Asp Pro Asp Asn Ile Ile Leu Val Gly Ser Pro 
                165                 170                 175     


His Trp Asp Gln Asp Leu His Leu Val Ala Glu Asp Pro Ile Arg Asp 
            180                 185                 190         


Val Ser Asn Ile Met Tyr Thr Met His Phe Tyr Ala Ala Thr His Glu 
        195                 200                 205             


Ala Trp Leu Arg Asp Arg Thr Asp Glu Ala Ile Ala Lys Gly Ile Pro 
    210                 215                 220                 


Val Phe Val Ser Glu Cys Gly Gly Ser Glu Ala Asn Gly Asp Gly Arg 
225                 230                 235                 240 


Leu Gly Ile Glu Glu Trp Lys Thr Tyr Val Asp Trp Met Glu Ser Arg 
                245                 250                 255     


Lys Ile Ser Trp Val Ala Trp Ser Val Ser Asp Lys Asn Glu Thr Ala 
            260                 265                 270         


Ser Met Leu Leu Pro Arg Ala Ser Ala Asp Gly Asn Trp Thr Glu Asp 
        275                 280                 285             


Leu Leu Lys Pro Trp Gly Lys Leu Thr Arg Asn Ser Ile Arg Asn Ala 
    290                 295                 300                 


Asn Asp Glu Asn Pro Asp Ile 
305                 310     


<210>  14
<211>  311
<212>  PRT
<213>  Artificial sequence

<220>
<223>  Sequence representing double alanine substitutions made on SEQ ID
       NO 4 wherein cysteine 230 and 272 are replaced by alanine

<400>  14

Glu Asn Asn Arg Lys Thr Asp Tyr Arg Ser Ile Val Ala Gln Asn Gly 
1               5                   10                  15      


Arg Leu Gln Val Ile Gly Thr Gln Leu Ser Asn Glu Lys Gly Glu Pro 
            20                  25                  30          


Val Val Leu Arg Gly Ala Ser Leu Gly Trp His Asn Leu Trp Pro Arg 
        35                  40                  45              


Phe Tyr Asn Lys Asn Ala Val Gln Trp Leu Ala Asp Asp Trp Lys Cys 
    50                  55                  60                  


Thr Val Val Arg Ala Ala Met Gly Leu Glu Ile Glu Asp Asn Tyr Arg 
65                  70                  75                  80  


Glu Asn Pro Glu Phe Ala Leu Gln Cys Ile Thr Pro Val Ile Glu Ser 
                85                  90                  95      


Ala Ile Glu Asn Gly Ile Tyr Val Ile Ile Asp Phe His Ala His Asn 
            100                 105                 110         


Lys Tyr Thr Glu Glu Ala Lys Thr Phe Phe Ala Gly Met Ala Glu Lys 
        115                 120                 125             


Tyr Gly Glu Tyr Pro Asn Val Ile Tyr Glu Ile Trp Asn Glu Pro Asp 
    130                 135                 140                 


Tyr Phe Glu Trp Glu Glu Val Lys Thr Tyr Ser Glu Glu Val Ile Ala 
145                 150                 155                 160 


Val Ile Arg Ala Ile Asp Pro Asp Asn Ile Ile Leu Val Gly Ser Pro 
                165                 170                 175     


His Trp Asp Gln Asp Leu His Leu Val Ala Glu Asp Pro Ile Arg Asp 
            180                 185                 190         


Val Ser Asn Ile Met Tyr Thr Met His Phe Tyr Ala Ala Thr His Glu 
        195                 200                 205             


Ala Trp Leu Arg Asp Arg Thr Asp Glu Ala Ile Ala Lys Gly Ile Pro 
    210                 215                 220                 


Val Phe Val Ser Glu Ala Gly Gly Ser Glu Ala Asn Gly Asp Gly Arg 
225                 230                 235                 240 


Leu Gly Ile Glu Glu Trp Lys Thr Tyr Val Asp Trp Met Glu Ser Arg 
                245                 250                 255     


Lys Ile Ser Trp Val Ala Trp Ser Val Ser Asp Lys Asn Glu Thr Ala 
            260                 265                 270         


Ser Met Leu Leu Pro Arg Ala Ser Ala Asp Gly Asn Trp Thr Glu Asp 
        275                 280                 285             


Leu Leu Lys Pro Trp Gly Lys Leu Thr Arg Asn Ser Ile Arg Asn Ala 
    290                 295                 300                 


Asn Asp Glu Asn Pro Asp Ile 
305                 310     


<210>  15
<211>  311
<212>  PRT
<213>  Artificial sequence

<220>
<223>  Sequence representing triple alanine substitutions made on SEQ ID
       NO 4 wherein cysteine 64, 84 and 230 are replaced by alanine

<400>  15

Glu Asn Asn Arg Lys Thr Asp Tyr Arg Ser Ile Val Ala Gln Asn Gly 
1               5                   10                  15      


Arg Leu Gln Val Ile Gly Thr Gln Leu Ser Asn Glu Lys Gly Glu Pro 
            20                  25                  30          


Val Val Leu Arg Gly Ala Ser Leu Gly Trp His Asn Leu Trp Pro Arg 
        35                  40                  45              


Phe Tyr Asn Lys Asn Ala Val Gln Trp Leu Ala Asp Asp Trp Lys Ala 
    50                  55                  60                  


Thr Val Val Arg Ala Ala Met Gly Leu Glu Ile Glu Asp Asn Tyr Arg 
65                  70                  75                  80  


Glu Asn Pro Glu Phe Ala Leu Gln Ala Ile Thr Pro Val Ile Glu Ser 
                85                  90                  95      


Ala Ile Glu Asn Gly Ile Tyr Val Ile Ile Asp Phe His Ala His Asn 
            100                 105                 110         


Lys Tyr Thr Glu Glu Ala Lys Thr Phe Phe Ala Gly Met Ala Glu Lys 
        115                 120                 125             


Tyr Gly Glu Tyr Pro Asn Val Ile Tyr Glu Ile Trp Asn Glu Pro Asp 
    130                 135                 140                 


Tyr Phe Glu Trp Glu Glu Val Lys Thr Tyr Ser Glu Glu Val Ile Ala 
145                 150                 155                 160 


Val Ile Arg Ala Ile Asp Pro Asp Asn Ile Ile Leu Val Gly Ser Pro 
                165                 170                 175     


His Trp Asp Gln Asp Leu His Leu Val Ala Glu Asp Pro Ile Arg Asp 
            180                 185                 190         


Val Ser Asn Ile Met Tyr Thr Met His Phe Tyr Ala Ala Thr His Glu 
        195                 200                 205             


Ala Trp Leu Arg Asp Arg Thr Asp Glu Ala Ile Ala Lys Gly Ile Pro 
    210                 215                 220                 


Val Phe Val Ser Glu Ala Gly Gly Ser Glu Ala Asn Gly Asp Gly Arg 
225                 230                 235                 240 


Leu Gly Ile Glu Glu Trp Lys Thr Tyr Val Asp Trp Met Glu Ser Arg 
                245                 250                 255     


Lys Ile Ser Trp Val Ala Trp Ser Val Ser Asp Lys Asn Glu Thr Cys 
            260                 265                 270         


Ser Met Leu Leu Pro Arg Ala Ser Ala Asp Gly Asn Trp Thr Glu Asp 
        275                 280                 285             


Leu Leu Lys Pro Trp Gly Lys Leu Thr Arg Asn Ser Ile Arg Asn Ala 
    290                 295                 300                 


Asn Asp Glu Asn Pro Asp Ile 
305                 310     


<210>  16
<211>  311
<212>  PRT
<213>  Artificial sequence

<220>
<223>  Sequence representing triple alanine substitutions made on SEQ ID
       NO 4 wherein cysteine 64,230 and 272 are replaced by alanine

<400>  16

Glu Asn Asn Arg Lys Thr Asp Tyr Arg Ser Ile Val Ala Gln Asn Gly 
1               5                   10                  15      


Arg Leu Gln Val Ile Gly Thr Gln Leu Ser Asn Glu Lys Gly Glu Pro 
            20                  25                  30          


Val Val Leu Arg Gly Ala Ser Leu Gly Trp His Asn Leu Trp Pro Arg 
        35                  40                  45              


Phe Tyr Asn Lys Asn Ala Val Gln Trp Leu Ala Asp Asp Trp Lys Ala 
    50                  55                  60                  


Thr Val Val Arg Ala Ala Met Gly Leu Glu Ile Glu Asp Asn Tyr Arg 
65                  70                  75                  80  


Glu Asn Pro Glu Phe Ala Leu Gln Cys Ile Thr Pro Val Ile Glu Ser 
                85                  90                  95      


Ala Ile Glu Asn Gly Ile Tyr Val Ile Ile Asp Phe His Ala His Asn 
            100                 105                 110         


Lys Tyr Thr Glu Glu Ala Lys Thr Phe Phe Ala Gly Met Ala Glu Lys 
        115                 120                 125             


Tyr Gly Glu Tyr Pro Asn Val Ile Tyr Glu Ile Trp Asn Glu Pro Asp 
    130                 135                 140                 


Tyr Phe Glu Trp Glu Glu Val Lys Thr Tyr Ser Glu Glu Val Ile Ala 
145                 150                 155                 160 


Val Ile Arg Ala Ile Asp Pro Asp Asn Ile Ile Leu Val Gly Ser Pro 
                165                 170                 175     


His Trp Asp Gln Asp Leu His Leu Val Ala Glu Asp Pro Ile Arg Asp 
            180                 185                 190         


Val Ser Asn Ile Met Tyr Thr Met His Phe Tyr Ala Ala Thr His Glu 
        195                 200                 205             


Ala Trp Leu Arg Asp Arg Thr Asp Glu Ala Ile Ala Lys Gly Ile Pro 
    210                 215                 220                 


Val Phe Val Ser Glu Ala Gly Gly Ser Glu Ala Asn Gly Asp Gly Arg 
225                 230                 235                 240 


Leu Gly Ile Glu Glu Trp Lys Thr Tyr Val Asp Trp Met Glu Ser Arg 
                245                 250                 255     


Lys Ile Ser Trp Val Ala Trp Ser Val Ser Asp Lys Asn Glu Thr Ala 
            260                 265                 270         


Ser Met Leu Leu Pro Arg Ala Ser Ala Asp Gly Asn Trp Thr Glu Asp 
        275                 280                 285             


Leu Leu Lys Pro Trp Gly Lys Leu Thr Arg Asn Ser Ile Arg Asn Ala 
    290                 295                 300                 


Asn Asp Glu Asn Pro Asp Ile 
305                 310     


<210>  17
<211>  311
<212>  PRT
<213>  Artificial sequence

<220>
<223>  Sequence representing triple alanine substitutions made on SEQ ID
       NO 4 wherein cysteine 89, 230 and 272 are replaced by alanine

<400>  17

Glu Asn Asn Arg Lys Thr Asp Tyr Arg Ser Ile Val Ala Gln Asn Gly 
1               5                   10                  15      


Arg Leu Gln Val Ile Gly Thr Gln Leu Ser Asn Glu Lys Gly Glu Pro 
            20                  25                  30          


Val Val Leu Arg Gly Ala Ser Leu Gly Trp His Asn Leu Trp Pro Arg 
        35                  40                  45              


Phe Tyr Asn Lys Asn Ala Val Gln Trp Leu Ala Asp Asp Trp Lys Cys 
    50                  55                  60                  


Thr Val Val Arg Ala Ala Met Gly Leu Glu Ile Glu Asp Asn Tyr Arg 
65                  70                  75                  80  


Glu Asn Pro Glu Phe Ala Leu Gln Ala Ile Thr Pro Val Ile Glu Ser 
                85                  90                  95      


Ala Ile Glu Asn Gly Ile Tyr Val Ile Ile Asp Phe His Ala His Asn 
            100                 105                 110         


Lys Tyr Thr Glu Glu Ala Lys Thr Phe Phe Ala Gly Met Ala Glu Lys 
        115                 120                 125             


Tyr Gly Glu Tyr Pro Asn Val Ile Tyr Glu Ile Trp Asn Glu Pro Asp 
    130                 135                 140                 


Tyr Phe Glu Trp Glu Glu Val Lys Thr Tyr Ser Glu Glu Val Ile Ala 
145                 150                 155                 160 


Val Ile Arg Ala Ile Asp Pro Asp Asn Ile Ile Leu Val Gly Ser Pro 
                165                 170                 175     


His Trp Asp Gln Asp Leu His Leu Val Ala Glu Asp Pro Ile Arg Asp 
            180                 185                 190         


Val Ser Asn Ile Met Tyr Thr Met His Phe Tyr Ala Ala Thr His Glu 
        195                 200                 205             


Ala Trp Leu Arg Asp Arg Thr Asp Glu Ala Ile Ala Lys Gly Ile Pro 
    210                 215                 220                 


Val Phe Val Ser Glu Ala Gly Gly Ser Glu Ala Asn Gly Asp Gly Arg 
225                 230                 235                 240 


Leu Gly Ile Glu Glu Trp Lys Thr Tyr Val Asp Trp Met Glu Ser Arg 
                245                 250                 255     


Lys Ile Ser Trp Val Ala Trp Ser Val Ser Asp Lys Asn Glu Thr Ala 
            260                 265                 270         


Ser Met Leu Leu Pro Arg Ala Ser Ala Asp Gly Asn Trp Thr Glu Asp 
        275                 280                 285             


Leu Leu Lys Pro Trp Gly Lys Leu Thr Arg Asn Ser Ile Arg Asn Ala 
    290                 295                 300                 


Asn Asp Glu Asn Pro Asp Ile 
305                 310     


<210>  18
<211>  311
<212>  PRT
<213>  Artificial sequence

<220>
<223>  Sequence representing triple alanine substitutions made on SEQ ID
       NO 4 wherein cysteine 64,89 and 272 are replaced by alanine

<400>  18

Glu Asn Asn Arg Lys Thr Asp Tyr Arg Ser Ile Val Ala Gln Asn Gly 
1               5                   10                  15      


Arg Leu Gln Val Ile Gly Thr Gln Leu Ser Asn Glu Lys Gly Glu Pro 
            20                  25                  30          


Val Val Leu Arg Gly Ala Ser Leu Gly Trp His Asn Leu Trp Pro Arg 
        35                  40                  45              


Phe Tyr Asn Lys Asn Ala Val Gln Trp Leu Ala Asp Asp Trp Lys Ala 
    50                  55                  60                  


Thr Val Val Arg Ala Ala Met Gly Leu Glu Ile Glu Asp Asn Tyr Arg 
65                  70                  75                  80  


Glu Asn Pro Glu Phe Ala Leu Gln Ala Ile Thr Pro Val Ile Glu Ser 
                85                  90                  95      


Ala Ile Glu Asn Gly Ile Tyr Val Ile Ile Asp Phe His Ala His Asn 
            100                 105                 110         


Lys Tyr Thr Glu Glu Ala Lys Thr Phe Phe Ala Gly Met Ala Glu Lys 
        115                 120                 125             


Tyr Gly Glu Tyr Pro Asn Val Ile Tyr Glu Ile Trp Asn Glu Pro Asp 
    130                 135                 140                 


Tyr Phe Glu Trp Glu Glu Val Lys Thr Tyr Ser Glu Glu Val Ile Ala 
145                 150                 155                 160 


Val Ile Arg Ala Ile Asp Pro Asp Asn Ile Ile Leu Val Gly Ser Pro 
                165                 170                 175     


His Trp Asp Gln Asp Leu His Leu Val Ala Glu Asp Pro Ile Arg Asp 
            180                 185                 190         


Val Ser Asn Ile Met Tyr Thr Met His Phe Tyr Ala Ala Thr His Glu 
        195                 200                 205             


Ala Trp Leu Arg Asp Arg Thr Asp Glu Ala Ile Ala Lys Gly Ile Pro 
    210                 215                 220                 


Val Phe Val Ser Glu Cys Gly Gly Ser Glu Ala Asn Gly Asp Gly Arg 
225                 230                 235                 240 


Leu Gly Ile Glu Glu Trp Lys Thr Tyr Val Asp Trp Met Glu Ser Arg 
                245                 250                 255     


Lys Ile Ser Trp Val Ala Trp Ser Val Ser Asp Lys Asn Glu Thr Ala 
            260                 265                 270         


Ser Met Leu Leu Pro Arg Ala Ser Ala Asp Gly Asn Trp Thr Glu Asp 
        275                 280                 285             


Leu Leu Lys Pro Trp Gly Lys Leu Thr Arg Asn Ser Ile Arg Asn Ala 
    290                 295                 300                 


Asn Asp Glu Asn Pro Asp Ile 
305                 310     


<210>  19
<211>  311
<212>  PRT
<213>  Artificial sequence

<220>
<223>  Sequence representing quadruple alanine substitutions made on SEQ
       ID NO 4 wherein cysteine 64,89,230 and 272 are replaced by 
       alanine

<400>  19

Glu Asn Asn Arg Lys Thr Asp Tyr Arg Ser Ile Val Ala Gln Asn Gly 
1               5                   10                  15      


Arg Leu Gln Val Ile Gly Thr Gln Leu Ser Asn Glu Lys Gly Glu Pro 
            20                  25                  30          


Val Val Leu Arg Gly Ala Ser Leu Gly Trp His Asn Leu Trp Pro Arg 
        35                  40                  45              


Phe Tyr Asn Lys Asn Ala Val Gln Trp Leu Ala Asp Asp Trp Lys Ala 
    50                  55                  60                  


Thr Val Val Arg Ala Ala Met Gly Leu Glu Ile Glu Asp Asn Tyr Arg 
65                  70                  75                  80  


Glu Asn Pro Glu Phe Ala Leu Gln Ala Ile Thr Pro Val Ile Glu Ser 
                85                  90                  95      


Ala Ile Glu Asn Gly Ile Tyr Val Ile Ile Asp Phe His Ala His Asn 
            100                 105                 110         


Lys Tyr Thr Glu Glu Ala Lys Thr Phe Phe Ala Gly Met Ala Glu Lys 
        115                 120                 125             


Tyr Gly Glu Tyr Pro Asn Val Ile Tyr Glu Ile Trp Asn Glu Pro Asp 
    130                 135                 140                 


Tyr Phe Glu Trp Glu Glu Val Lys Thr Tyr Ser Glu Glu Val Ile Ala 
145                 150                 155                 160 


Val Ile Arg Ala Ile Asp Pro Asp Asn Ile Ile Leu Val Gly Ser Pro 
                165                 170                 175     


His Trp Asp Gln Asp Leu His Leu Val Ala Glu Asp Pro Ile Arg Asp 
            180                 185                 190         


Val Ser Asn Ile Met Tyr Thr Met His Phe Tyr Ala Ala Thr His Glu 
        195                 200                 205             


Ala Trp Leu Arg Asp Arg Thr Asp Glu Ala Ile Ala Lys Gly Ile Pro 
    210                 215                 220                 


Val Phe Val Ser Glu Ala Gly Gly Ser Glu Ala Asn Gly Asp Gly Arg 
225                 230                 235                 240 


Leu Gly Ile Glu Glu Trp Lys Thr Tyr Val Asp Trp Met Glu Ser Arg 
                245                 250                 255     


Lys Ile Ser Trp Val Ala Trp Ser Val Ser Asp Lys Asn Glu Thr Ala 
            260                 265                 270         


Ser Met Leu Leu Pro Arg Ala Ser Ala Asp Gly Asn Trp Thr Glu Asp 
        275                 280                 285             


Leu Leu Lys Pro Trp Gly Lys Leu Thr Arg Asn Ser Ile Arg Asn Ala 
    290                 295                 300                 


Asn Asp Glu Asn Pro Asp Ile 
305                 310     


<210>  20
<211>  33
<212>  DNA
<213>  Artificial sequence

<220>
<223>  forward primer for amplification of full length gene


<220>
<221>  primer
<222>  (1)..(33)
<223>  synthetically generated primer for amplification of full length 
       gene

<300>
<313>  (1)..(33)

<400>  20
aatatacata tgaagaaaaa ctcaatcatt ctc                                    33


<210>  21
<211>  33
<212>  DNA
<213>  Artificial sequence

<220>
<223>  synthetically generated primer


<220>
<221>  primer
<222>  (1)..(33)
<223>  synthetically generated primer for amplification of full length 
       gene

<300>
<313>  (1)..(33)

<400>  21
aatataggat cctcagatat ccgggttttc atc                                    33


<210>  22
<211>  35
<212>  DNA
<213>  artificial sequence

<220>
<223>  synthetically generated primer


<220>
<221>  primer
<222>  (1)..(35)
<223>  synthetically generated forward primer for amplication of 
       truncated gene

<400>  22
aatatacata tggaaaacaa caggaaaacg gacta                                  35


<210>  23
<211>  24
<212>  DNA
<213>  artificial sequence

<220>
<223>  synthetically generated primer


<220>
<221>  primer
<222>  (1)..(24)
<223>  synthetically generated forward primer for Cys64Ala

<400>  23
cgattggaaa gcgacggtcg tcag                                              24


<210>  24
<211>  24
<212>  DNA
<213>  Artificial sequence

<220>
<223>  synthetically generated primer


<220>
<221>  primer
<222>  (1)..(24)
<223>  synthetically generated reverse primer for Cys64Ala

<400>  24
ctgacgaccg tcgctttcca atcg                                              24


<210>  25
<211>  28
<212>  DNA
<213>  artificial sequence

<220>
<223>  synthetically generated primer


<220>
<221>  primer
<222>  (1)..(28)
<223>  synthetically generated forward primer for Cys89Ala

<400>  25
gaatttgcct tacaggcgat cacccctg                                          28


<210>  26
<211>  28
<212>  DNA
<213>  artificial sequence

<220>
<223>  synthetically generated primer


<220>
<221>  primer
<222>  (1)..(28)
<223>  synthetically generated reverse primer for Cys89Ala

<400>  26
caggggtgat cgcctgtaag gcaaattc                                          28


<210>  27
<211>  21
<212>  DNA
<213>  Aartificial sequence


<220>
<221>  primer
<222>  (1)..(21)
<223>  syntheically generated forward primer sequence for Cys230Ala

<400>  27
cgtatcggag gcgggcggct c                                                 21


<210>  28
<211>  21
<212>  DNA
<213>  Artificial sequence

<220>
<223>  synthetically generated primer


<220>
<221>  primer
<222>  (1)..(21)
<223>  syntheically generated forward primer sequence for Cys230Ala

<220>
<221>  primer
<222>  (1)..(21)
<223>  syntheically generated reverse primer sequence for Cys230Ala

<400>  28
gagccgcccg cctccgatac g                                                 21


<210>  29
<211>  25
<212>  DNA
<213>  artificial sequence

<220>
<223>  synthetically generated primer


<220>
<221>  primer1
<222>  (1)..(25)
<223>  syntheically generated forward primer sequence for Cys272Ala

<400>  29
gaacgaaacc gcgtccatgc tgctc                                             25


<210>  30
<211>  25
<212>  DNA
<213>  artificial sequence

<220>
<223>  synthetically generated primer


<220>
<221>  primer
<222>  (1)..(25)
<223>  syntheically generated reverse primer sequence for Cys272Ala

<400>  30
gagcagcatg gacgcggttt cgttc                                             25


<210>  31
<211>  936
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Nucleotide sequence representing single alanine substitution made
       on SEQ ID N0: 3 wherein the codon for cysteine at position 190 is
       replaced by alanine

<400>  31
gaaaacaaca ggaaaacgga ctaccgttcc atcgtagcgc agaacggccg tttacaggtg       60

atcggcacac aattgagcaa tgaaaaaggc gaacccgtcg ttttacgggg agccagcctc      120

ggatggcaca acctctggcc ccgcttctat aacaagaacg cggtgcaatg gctggccgac      180

gattggaaag cgacggtcgt cagggctgca atgggcttgg aaattgaaga caactaccgg      240

gaaaatcccg aatttgcctt acagtgcatc acccctgtca tcgaatcggc tattgaaaac      300

ggaatctatg tgattatcga cttccacgca cacaacaaat acaccgaaga agccaaaaca      360

ttctttgccg ggatggccga aaaatacggg gaatatccga acgtgatcta tgaaatctgg      420

aacgaacccg attatttcga atgggaagaa gtaaaaacct attcggaaga agtgatcgcc      480

gtcatccgcg cgatcgaccc cgacaatatt atattggtcg gcagccccca ttgggaccag      540

gacctgcatc tggtagccga agacccgatc cgggatgtaa gcaacatcat gtacaccatg      600

catttttacg ccgcgaccca cgaggcctgg ctgcgtgacc ggaccgacga ggcgattgcc      660

aaaggaattc ccgttttcgt atcggagtgc ggcggctcgg aagccaatgg cgacggacgg      720

ttaggcatag aagaatggaa aacctacgtc gattggatgg agagtcggaa aataagctgg      780

gtggcctggt ccgtttccga caagaacgaa acctgctcca tgctgctccc ccgcgcctct      840

gccgatggca actggacgga agacctgctc aagccttggg gaaaactgac gcgtaattct      900

atccggaacg cgaatgatga aaacccggat atctga                                936


<210>  32
<211>  936
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Nucleotide sequence representing single alanine substitution made
       on SEQ ID N0: 3 wherein the codon for cysteine at position 265 is
       replaced by alanine

<400>  32
gaaaacaaca ggaaaacgga ctaccgttcc atcgtagcgc agaacggccg tttacaggtg       60

atcggcacac aattgagcaa tgaaaaaggc gaacccgtcg ttttacgggg agccagcctc      120

ggatggcaca acctctggcc ccgcttctat aacaagaacg cggtgcaatg gctggccgac      180

gattggaaat gcacggtcgt cagggctgca atgggcttgg aaattgaaga caactaccgg      240

gaaaatcccg aatttgcctt acaggcgatc acccctgtca tcgaatcggc tattgaaaac      300

ggaatctatg tgattatcga cttccacgca cacaacaaat acaccgaaga agccaaaaca      360

ttctttgccg ggatggccga aaaatacggg gaatatccga acgtgatcta tgaaatctgg      420

aacgaacccg attatttcga atgggaagaa gtaaaaacct attcggaaga agtgatcgcc      480

gtcatccgcg cgatcgaccc cgacaatatt atattggtcg gcagccccca ttgggaccag      540

gacctgcatc tggtagccga agacccgatc cgggatgtaa gcaacatcat gtacaccatg      600

catttttacg ccgcgaccca cgaggcctgg ctgcgtgacc ggaccgacga ggcgattgcc      660

aaaggaattc ccgttttcgt atcggagtgc ggcggctcgg aagccaatgg cgacggacgg      720

ttaggcatag aagaatggaa aacctacgtc gattggatgg agagtcggaa aataagctgg      780

gtggcctggt ccgtttccga caagaacgaa acctgctcca tgctgctccc ccgcgcctct      840

gccgatggca actggacgga agacctgctc aagccttggg gaaaactgac gcgtaattct      900

atccggaacg cgaatgatga aaacccggat atctga                                936


<210>  33
<211>  936
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Nucleotide sequence representing single alanine substitution made
       on SEQ ID N0: 3 wherein the codon for cysteine at position 688 is
       replaced by alanine

<400>  33
gaaaacaaca ggaaaacgga ctaccgttcc atcgtagcgc agaacggccg tttacaggtg       60

atcggcacac aattgagcaa tgaaaaaggc gaacccgtcg ttttacgggg agccagcctc      120

ggatggcaca acctctggcc ccgcttctat aacaagaacg cggtgcaatg gctggccgac      180

gattggaaat gcacggtcgt cagggctgca atgggcttgg aaattgaaga caactaccgg      240

gaaaatcccg aatttgcctt acagtgcatc acccctgtca tcgaatcggc tattgaaaac      300

ggaatctatg tgattatcga cttccacgca cacaacaaat acaccgaaga agccaaaaca      360

ttctttgccg ggatggccga aaaatacggg gaatatccga acgtgatcta tgaaatctgg      420

aacgaacccg attatttcga atgggaagaa gtaaaaacct attcggaaga agtgatcgcc      480

gtcatccgcg cgatcgaccc cgacaatatt atattggtcg gcagccccca ttgggaccag      540

gacctgcatc tggtagccga agacccgatc cgggatgtaa gcaacatcat gtacaccatg      600

catttttacg ccgcgaccca cgaggcctgg ctgcgtgacc ggaccgacga ggcgattgcc      660

aaaggaattc ccgttttcgt atcggaggcg ggcggctcgg aagccaatgg cgacggacgg      720

ttaggcatag aagaatggaa aacctacgtc gattggatgg agagtcggaa aataagctgg      780

gtggcctggt ccgtttccga caagaacgaa acctgctcca tgctgctccc ccgcgcctct      840

gccgatggca actggacgga agacctgctc aagccttggg gaaaactgac gcgtaattct      900

atccggaacg cgaatgatga aaacccggat atctga                                936


<210>  34
<211>  936
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Nucleotide sequence representing single alanine substitution made
       on SEQ ID N0: 3 wherein the codon for cysteine at position 814 is
       replaced by alanine

<400>  34
gaaaacaaca ggaaaacgga ctaccgttcc atcgtagcgc agaacggccg tttacaggtg       60

atcggcacac aattgagcaa tgaaaaaggc gaacccgtcg ttttacgggg agccagcctc      120

ggatggcaca acctctggcc ccgcttctat aacaagaacg cggtgcaatg gctggccgac      180

gattggaaat gcacggtcgt cagggctgca atgggcttgg aaattgaaga caactaccgg      240

gaaaatcccg aatttgcctt acagtgcatc acccctgtca tcgaatcggc tattgaaaac      300

ggaatctatg tgattatcga cttccacgca cacaacaaat acaccgaaga agccaaaaca      360

ttctttgccg ggatggccga aaaatacggg gaatatccga acgtgatcta tgaaatctgg      420

aacgaacccg attatttcga atgggaagaa gtaaaaacct attcggaaga agtgatcgcc      480

gtcatccgcg cgatcgaccc cgacaatatt atattggtcg gcagccccca ttgggaccag      540

gacctgcatc tggtagccga agacccgatc cgggatgtaa gcaacatcat gtacaccatg      600

catttttacg ccgcgaccca cgaggcctgg ctgcgtgacc ggaccgacga ggcgattgcc      660

aaaggaattc ccgttttcgt atcggagtgc ggcggctcgg aagccaatgg cgacggacgg      720

ttaggcatag aagaatggaa aacctacgtc gattggatgg agagtcggaa aataagctgg      780

gtggcctggt ccgtttccga caagaacgaa accgcgtcca tgctgctccc ccgcgcctct      840

gccgatggca actggacgga agacctgctc aagccttggg gaaaactgac gcgtaattct      900

atccggaacg cgaatgatga aaacccggat atctga                                936


<210>  35
<211>  936
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Nucleotide sequence representing double alanine substitutions 
       made on SEQ ID N0: 3 wherein the codon for cysteine at positions 
       190 and 265 are replaced by alanine

<400>  35
gaaaacaaca ggaaaacgga ctaccgttcc atcgtagcgc agaacggccg tttacaggtg       60

atcggcacac aattgagcaa tgaaaaaggc gaacccgtcg ttttacgggg agccagcctc      120

ggatggcaca acctctggcc ccgcttctat aacaagaacg cggtgcaatg gctggccgac      180

gattggaaag cgacggtcgt cagggctgca atgggcttgg aaattgaaga caactaccgg      240

gaaaatcccg aatttgcctt acaggcgatc acccctgtca tcgaatcggc tattgaaaac      300

ggaatctatg tgattatcga cttccacgca cacaacaaat acaccgaaga agccaaaaca      360

ttctttgccg ggatggccga aaaatacggg gaatatccga acgtgatcta tgaaatctgg      420

aacgaacccg attatttcga atgggaagaa gtaaaaacct attcggaaga agtgatcgcc      480

gtcatccgcg cgatcgaccc cgacaatatt atattggtcg gcagccccca ttgggaccag      540

gacctgcatc tggtagccga agacccgatc cgggatgtaa gcaacatcat gtacaccatg      600

catttttacg ccgcgaccca cgaggcctgg ctgcgtgacc ggaccgacga ggcgattgcc      660

aaaggaattc ccgttttcgt atcggagtgc ggcggctcgg aagccaatgg cgacggacgg      720

ttaggcatag aagaatggaa aacctacgtc gattggatgg agagtcggaa aataagctgg      780

gtggcctggt ccgtttccga caagaacgaa acctgctcca tgctgctccc ccgcgcctct      840

gccgatggca actggacgga agacctgctc aagccttggg gaaaactgac gcgtaattct      900

atccggaacg cgaatgatga aaacccggat atctga                                936


<210>  36
<211>  936
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Nucleotide sequence representing double alanine substitutions 
       made on SEQ ID N0: 3 wherein the codon for cysteine at positions 
       190 and 688 are replaced by alanine

<400>  36
gaaaacaaca ggaaaacgga ctaccgttcc atcgtagcgc agaacggccg tttacaggtg       60

atcggcacac aattgagcaa tgaaaaaggc gaacccgtcg ttttacgggg agccagcctc      120

ggatggcaca acctctggcc ccgcttctat aacaagaacg cggtgcaatg gctggccgac      180

gattggaaag cgacggtcgt cagggctgca atgggcttgg aaattgaaga caactaccgg      240

gaaaatcccg aatttgcctt acagtgcatc acccctgtca tcgaatcggc tattgaaaac      300

ggaatctatg tgattatcga cttccacgca cacaacaaat acaccgaaga agccaaaaca      360

ttctttgccg ggatggccga aaaatacggg gaatatccga acgtgatcta tgaaatctgg      420

aacgaacccg attatttcga atgggaagaa gtaaaaacct attcggaaga agtgatcgcc      480

gtcatccgcg cgatcgaccc cgacaatatt atattggtcg gcagccccca ttgggaccag      540

gacctgcatc tggtagccga agacccgatc cgggatgtaa gcaacatcat gtacaccatg      600

catttttacg ccgcgaccca cgaggcctgg ctgcgtgacc ggaccgacga ggcgattgcc      660

aaaggaattc ccgttttcgt atcggaggcg ggcggctcgg aagccaatgg cgacggacgg      720

ttaggcatag aagaatggaa aacctacgtc gattggatgg agagtcggaa aataagctgg      780

gtggcctggt ccgtttccga caagaacgaa acctgctcca tgctgctccc ccgcgcctct      840

gccgatggca actggacgga agacctgctc aagccttggg gaaaactgac gcgtaattct      900

atccggaacg cgaatgatga aaacccggat atctga                                936


<210>  37
<211>  936
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Nucleotide sequence representing double alanine substitutions 
       made on SEQ ID N0: 3 wherein the codon for cysteine at positions 
       190 and 814 are replaced by alanine

<400>  37
gaaaacaaca ggaaaacgga ctaccgttcc atcgtagcgc agaacggccg tttacaggtg       60

atcggcacac aattgagcaa tgaaaaaggc gaacccgtcg ttttacgggg agccagcctc      120

ggatggcaca acctctggcc ccgcttctat aacaagaacg cggtgcaatg gctggccgac      180

gattggaaag cgacggtcgt cagggctgca atgggcttgg aaattgaaga caactaccgg      240

gaaaatcccg aatttgcctt acagtgcatc acccctgtca tcgaatcggc tattgaaaac      300

ggaatctatg tgattatcga cttccacgca cacaacaaat acaccgaaga agccaaaaca      360

ttctttgccg ggatggccga aaaatacggg gaatatccga acgtgatcta tgaaatctgg      420

aacgaacccg attatttcga atgggaagaa gtaaaaacct attcggaaga agtgatcgcc      480

gtcatccgcg cgatcgaccc cgacaatatt atattggtcg gcagccccca ttgggaccag      540

gacctgcatc tggtagccga agacccgatc cgggatgtaa gcaacatcat gtacaccatg      600

catttttacg ccgcgaccca cgaggcctgg ctgcgtgacc ggaccgacga ggcgattgcc      660

aaaggaattc ccgttttcgt atcggagtgc ggcggctcgg aagccaatgg cgacggacgg      720

ttaggcatag aagaatggaa aacctacgtc gattggatgg agagtcggaa aataagctgg      780

gtggcctggt ccgtttccga caagaacgaa accgcgtcca tgctgctccc ccgcgcctct      840

gccgatggca actggacgga agacctgctc aagccttggg gaaaactgac gcgtaattct      900

atccggaacg cgaatgatga aaacccggat atctga                                936


<210>  38
<211>  936
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Nucleotide sequence representing double alanine substitutions 
       made on SEQ ID N0: 3 wherein the codon for cysteine at positions 
       265 and 688 are replaced by alanine

<400>  38
gaaaacaaca ggaaaacgga ctaccgttcc atcgtagcgc agaacggccg tttacaggtg       60

atcggcacac aattgagcaa tgaaaaaggc gaacccgtcg ttttacgggg agccagcctc      120

ggatggcaca acctctggcc ccgcttctat aacaagaacg cggtgcaatg gctggccgac      180

gattggaaat gcacggtcgt cagggctgca atgggcttgg aaattgaaga caactaccgg      240

gaaaatcccg aatttgcctt acaggcgatc acccctgtca tcgaatcggc tattgaaaac      300

ggaatctatg tgattatcga cttccacgca cacaacaaat acaccgaaga agccaaaaca      360

ttctttgccg ggatggccga aaaatacggg gaatatccga acgtgatcta tgaaatctgg      420

aacgaacccg attatttcga atgggaagaa gtaaaaacct attcggaaga agtgatcgcc      480

gtcatccgcg cgatcgaccc cgacaatatt atattggtcg gcagccccca ttgggaccag      540

gacctgcatc tggtagccga agacccgatc cgggatgtaa gcaacatcat gtacaccatg      600

catttttacg ccgcgaccca cgaggcctgg ctgcgtgacc ggaccgacga ggcgattgcc      660

aaaggaattc ccgttttcgt atcggaggcg ggcggctcgg aagccaatgg cgacggacgg      720

ttaggcatag aagaatggaa aacctacgtc gattggatgg agagtcggaa aataagctgg      780

gtggcctggt ccgtttccga caagaacgaa acctgctcca tgctgctccc ccgcgcctct      840

gccgatggca actggacgga agacctgctc aagccttggg gaaaactgac gcgtaattct      900

atccggaacg cgaatgatga aaacccggat atctga                                936


<210>  39
<211>  936
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Nucleotide sequence representing double alanine substitutions 
       made on SEQ ID N0: 3 wherein the codon for cysteine at positions 
       265 and 814 are replaced by alanine

<400>  39
gaaaacaaca ggaaaacgga ctaccgttcc atcgtagcgc agaacggccg tttacaggtg       60

atcggcacac aattgagcaa tgaaaaaggc gaacccgtcg ttttacgggg agccagcctc      120

ggatggcaca acctctggcc ccgcttctat aacaagaacg cggtgcaatg gctggccgac      180

gattggaaat gcacggtcgt cagggctgca atgggcttgg aaattgaaga caactaccgg      240

gaaaatcccg aatttgcctt acaggcgatc acccctgtca tcgaatcggc tattgaaaac      300

ggaatctatg tgattatcga cttccacgca cacaacaaat acaccgaaga agccaaaaca      360

ttctttgccg ggatggccga aaaatacggg gaatatccga acgtgatcta tgaaatctgg      420

aacgaacccg attatttcga atgggaagaa gtaaaaacct attcggaaga agtgatcgcc      480

gtcatccgcg cgatcgaccc cgacaatatt atattggtcg gcagccccca ttgggaccag      540

gacctgcatc tggtagccga agacccgatc cgggatgtaa gcaacatcat gtacaccatg      600

catttttacg ccgcgaccca cgaggcctgg ctgcgtgacc ggaccgacga ggcgattgcc      660

aaaggaattc ccgttttcgt atcggagtgc ggcggctcgg aagccaatgg cgacggacgg      720

ttaggcatag aagaatggaa aacctacgtc gattggatgg agagtcggaa aataagctgg      780

gtggcctggt ccgtttccga caagaacgaa accgcgtcca tgctgctccc ccgcgcctct      840

gccgatggca actggacgga agacctgctc aagccttggg gaaaactgac gcgtaattct      900

atccggaacg cgaatgatga aaacccggat atctga                                936


<210>  40
<211>  936
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Nucleotide sequence representing double alanine substitutions 
       made on SEQ ID N0: 3 wherein the codon for cysteine at positions 
       688 and 814 are replaced by alanine

<400>  40
gaaaacaaca ggaaaacgga ctaccgttcc atcgtagcgc agaacggccg tttacaggtg       60

atcggcacac aattgagcaa tgaaaaaggc gaacccgtcg ttttacgggg agccagcctc      120

ggatggcaca acctctggcc ccgcttctat aacaagaacg cggtgcaatg gctggccgac      180

gattggaaat gcacggtcgt cagggctgca atgggcttgg aaattgaaga caactaccgg      240

gaaaatcccg aatttgcctt acagtgcatc acccctgtca tcgaatcggc tattgaaaac      300

ggaatctatg tgattatcga cttccacgca cacaacaaat acaccgaaga agccaaaaca      360

ttctttgccg ggatggccga aaaatacggg gaatatccga acgtgatcta tgaaatctgg      420

aacgaacccg attatttcga atgggaagaa gtaaaaacct attcggaaga agtgatcgcc      480

gtcatccgcg cgatcgaccc cgacaatatt atattggtcg gcagccccca ttgggaccag      540

gacctgcatc tggtagccga agacccgatc cgggatgtaa gcaacatcat gtacaccatg      600

catttttacg ccgcgaccca cgaggcctgg ctgcgtgacc ggaccgacga ggcgattgcc      660

aaaggaattc ccgttttcgt atcggaggcg ggcggctcgg aagccaatgg cgacggacgg      720

ttaggcatag aagaatggaa aacctacgtc gattggatgg agagtcggaa aataagctgg      780

gtggcctggt ccgtttccga caagaacgaa accgcgtcca tgctgctccc ccgcgcctct      840

gccgatggca actggacgga agacctgctc aagccttggg gaaaactgac gcgtaattct      900

atccggaacg cgaatgatga aaacccggat atctga                                936


<210>  41
<211>  936
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Nucleotide sequence representing triple alanine substitutions 
       made on SEQ ID N0: 3 wherein the codon for cysteine at positions 
       190, 265 and 688 are replaced by alanine

<400>  41
gaaaacaaca ggaaaacgga ctaccgttcc atcgtagcgc agaacggccg tttacaggtg       60

atcggcacac aattgagcaa tgaaaaaggc gaacccgtcg ttttacgggg agccagcctc      120

ggatggcaca acctctggcc ccgcttctat aacaagaacg cggtgcaatg gctggccgac      180

gattggaaag cgacggtcgt cagggctgca atgggcttgg aaattgaaga caactaccgg      240

gaaaatcccg aatttgcctt acaggcgatc acccctgtca tcgaatcggc tattgaaaac      300

ggaatctatg tgattatcga cttccacgca cacaacaaat acaccgaaga agccaaaaca      360

ttctttgccg ggatggccga aaaatacggg gaatatccga acgtgatcta tgaaatctgg      420

aacgaacccg attatttcga atgggaagaa gtaaaaacct attcggaaga agtgatcgcc      480

gtcatccgcg cgatcgaccc cgacaatatt atattggtcg gcagccccca ttgggaccag      540

gacctgcatc tggtagccga agacccgatc cgggatgtaa gcaacatcat gtacaccatg      600

catttttacg ccgcgaccca cgaggcctgg ctgcgtgacc ggaccgacga ggcgattgcc      660

aaaggaattc ccgttttcgt atcggaggcg ggcggctcgg aagccaatgg cgacggacgg      720

ttaggcatag aagaatggaa aacctacgtc gattggatgg agagtcggaa aataagctgg      780

gtggcctggt ccgtttccga caagaacgaa acctgctcca tgctgctccc ccgcgcctct      840

gccgatggca actggacgga agacctgctc aagccttggg gaaaactgac gcgtaattct      900

atccggaacg cgaatgatga aaacccggat atctga                                936


<210>  42
<211>  936
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Nucleotide sequence representing triple alanine substitutions 
       made on SEQ ID N0: 3 wherein the codon for cysteine at positions 
       190, 688 and 814 are replaced by alanine

<400>  42
gaaaacaaca ggaaaacgga ctaccgttcc atcgtagcgc agaacggccg tttacaggtg       60

atcggcacac aattgagcaa tgaaaaaggc gaacccgtcg ttttacgggg agccagcctc      120

ggatggcaca acctctggcc ccgcttctat aacaagaacg cggtgcaatg gctggccgac      180

gattggaaag cgacggtcgt cagggctgca atgggcttgg aaattgaaga caactaccgg      240

gaaaatcccg aatttgcctt acagtgcatc acccctgtca tcgaatcggc tattgaaaac      300

ggaatctatg tgattatcga cttccacgca cacaacaaat acaccgaaga agccaaaaca      360

ttctttgccg ggatggccga aaaatacggg gaatatccga acgtgatcta tgaaatctgg      420

aacgaacccg attatttcga atgggaagaa gtaaaaacct attcggaaga agtgatcgcc      480

gtcatccgcg cgatcgaccc cgacaatatt atattggtcg gcagccccca ttgggaccag      540

gacctgcatc tggtagccga agacccgatc cgggatgtaa gcaacatcat gtacaccatg      600

catttttacg ccgcgaccca cgaggcctgg ctgcgtgacc ggaccgacga ggcgattgcc      660

aaaggaattc ccgttttcgt atcggaggcg ggcggctcgg aagccaatgg cgacggacgg      720

ttaggcatag aagaatggaa aacctacgtc gattggatgg agagtcggaa aataagctgg      780

gtggcctggt ccgtttccga caagaacgaa accgcgtcca tgctgctccc ccgcgcctct      840

gccgatggca actggacgga agacctgctc aagccttggg gaaaactgac gcgtaattct      900

atccggaacg cgaatgatga aaacccggat atctga                                936


<210>  43
<211>  936
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence representing triple alanine substitutions 
       made on SEQ ID N0: 3 wherein the codon for cysteine at positions 
       265, 688 and 814 are replaced by alanine

<400>  43
gaaaacaaca ggaaaacgga ctaccgttcc atcgtagcgc agaacggccg tttacaggtg       60

atcggcacac aattgagcaa tgaaaaaggc gaacccgtcg ttttacgggg agccagcctc      120

ggatggcaca acctctggcc ccgcttctat aacaagaacg cggtgcaatg gctggccgac      180

gattggaaat gcacggtcgt cagggctgca atgggcttgg aaattgaaga caactaccgg      240

gaaaatcccg aatttgcctt acaggcgatc acccctgtca tcgaatcggc tattgaaaac      300

ggaatctatg tgattatcga cttccacgca cacaacaaat acaccgaaga agccaaaaca      360

ttctttgccg ggatggccga aaaatacggg gaatatccga acgtgatcta tgaaatctgg      420

aacgaacccg attatttcga atgggaagaa gtaaaaacct attcggaaga agtgatcgcc      480

gtcatccgcg cgatcgaccc cgacaatatt atattggtcg gcagccccca ttgggaccag      540

gacctgcatc tggtagccga agacccgatc cgggatgtaa gcaacatcat gtacaccatg      600

catttttacg ccgcgaccca cgaggcctgg ctgcgtgacc ggaccgacga ggcgattgcc      660

aaaggaattc ccgttttcgt atcggaggcg ggcggctcgg aagccaatgg cgacggacgg      720

ttaggcatag aagaatggaa aacctacgtc gattggatgg agagtcggaa aataagctgg      780

gtggcctggt ccgtttccga caagaacgaa accgcgtcca tgctgctccc ccgcgcctct      840

gccgatggca actggacgga agacctgctc aagccttggg gaaaactgac gcgtaattct      900

atccggaacg cgaatgatga aaacccggat atctga                                936


<210>  44
<211>  936
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Nucleotide sequence representing triple alanine substitutions 
       made on SEQ ID N0: 3 wherein the codon for cysteine at positions 
       190, 265 and 814 are replaced by alanine

<400>  44
gaaaacaaca ggaaaacgga ctaccgttcc atcgtagcgc agaacggccg tttacaggtg       60

atcggcacac aattgagcaa tgaaaaaggc gaacccgtcg ttttacgggg agccagcctc      120

ggatggcaca acctctggcc ccgcttctat aacaagaacg cggtgcaatg gctggccgac      180

gattggaaag cgacggtcgt cagggctgca atgggcttgg aaattgaaga caactaccgg      240

gaaaatcccg aatttgcctt acaggcgatc acccctgtca tcgaatcggc tattgaaaac      300

ggaatctatg tgattatcga cttccacgca cacaacaaat acaccgaaga agccaaaaca      360

ttctttgccg ggatggccga aaaatacggg gaatatccga acgtgatcta tgaaatctgg      420

aacgaacccg attatttcga atgggaagaa gtaaaaacct attcggaaga agtgatcgcc      480

gtcatccgcg cgatcgaccc cgacaatatt atattggtcg gcagccccca ttgggaccag      540

gacctgcatc tggtagccga agacccgatc cgggatgtaa gcaacatcat gtacaccatg      600

catttttacg ccgcgaccca cgaggcctgg ctgcgtgacc ggaccgacga ggcgattgcc      660

aaaggaattc ccgttttcgt atcggagtgc ggcggctcgg aagccaatgg cgacggacgg      720

ttaggcatag aagaatggaa aacctacgtc gattggatgg agagtcggaa aataagctgg      780

gtggcctggt ccgtttccga caagaacgaa accgcgtcca tgctgctccc ccgcgcctct      840

gccgatggca actggacgga agacctgctc aagccttggg gaaaactgac gcgtaattct      900

atccggaacg cgaatgatga aaacccggat atctga                                936


<210>  45
<211>  936
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Nucleotide sequence representing quadruple alanine substitutions 
       made on SEQ ID N0: 3 wherein the codon for cysteine at positions 
       190, 265, 688 and 814 are replaced by alanine

<400>  45
gaaaacaaca ggaaaacgga ctaccgttcc atcgtagcgc agaacggccg tttacaggtg       60

atcggcacac aattgagcaa tgaaaaaggc gaacccgtcg ttttacgggg agccagcctc      120

ggatggcaca acctctggcc ccgcttctat aacaagaacg cggtgcaatg gctggccgac      180

gattggaaag cgacggtcgt cagggctgca atgggcttgg aaattgaaga caactaccgg      240

gaaaatcccg aatttgcctt acaggcgatc acccctgtca tcgaatcggc tattgaaaac      300

ggaatctatg tgattatcga cttccacgca cacaacaaat acaccgaaga agccaaaaca      360

ttctttgccg ggatggccga aaaatacggg gaatatccga acgtgatcta tgaaatctgg      420

aacgaacccg attatttcga atgggaagaa gtaaaaacct attcggaaga agtgatcgcc      480

gtcatccgcg cgatcgaccc cgacaatatt atattggtcg gcagccccca ttgggaccag      540

gacctgcatc tggtagccga agacccgatc cgggatgtaa gcaacatcat gtacaccatg      600

catttttacg ccgcgaccca cgaggcctgg ctgcgtgacc ggaccgacga ggcgattgcc      660

aaaggaattc ccgttttcgt atcggaggcg ggcggctcgg aagccaatgg cgacggacgg      720

ttaggcatag aagaatggaa aacctacgtc gattggatgg agagtcggaa aataagctgg      780

gtggcctggt ccgtttccga caagaacgaa accgcgtcca tgctgctccc ccgcgcctct      840

gccgatggca actggacgga agacctgctc aagccttggg gaaaactgac gcgtaattct      900

atccggaacg cgaatgatga aaacccggat atctga                                936



