                         SEQUENCE LISTING

<110>  EUROCLONE S.p.A.
       Universit degli Studi di Milano
 
<120>  Improved system for the expression of the receptor for the Advanced 
       Glycation End Products (AGE) and the Advanced Lipid Glycation End 
       Products (ALE) and applications thereof

<130>  @E0101605

<160>  23    

<170>  PatentIn version 3.5

<210>  1
<211>  762
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotidic sequence of His-VC1-StrepEc

<400>  1
atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat       60

atggctcaaa acatcacagc ccggattggc gagccactgg tgctgaagtg taagggggcc      120

cccaagaaac caccccagcg gctggaatgg aaactgaaca caggccggac agaagcttgg      180

aaggtcctgt ctccccaggg aggaggcccc tgggacagtg tggctcgtgt ccttcccaac      240

ggctccctct tccttccggc tgtcgggatc caggatgagg ggattttccg gtgccaggca      300

atgaacagga atggaaagga gaccaagtcc aactaccgag tccgtgtcta ccagattcct      360

gggaagccag aaattgtaga ttctgcctct gaactcacgg ctggtgttcc caataaggtg      420

gggacatgtg tgtcagaggg aagctaccct gcagggactc ttagctggca cttggatggg      480

aagcccctgg tgcctaatga gaagggagta tctgtgaagg aacagaccag gagacaccct      540

gagacagggc tcttcacact gcagtcggag ctaatggtga ccccagcccg gggaggagat      600

ccccgtccca ccttctcctg tagcttcagc ccaggccttc cccgacaccg ggccttgcgc      660

acagccccca tccagccccg tgtctgggag cctgtgcctc tggaggaggt ccaattggtg      720

gtggagggta gcgcttggcg tcacccgcag ttcggtggtt ag                         762


<210>  2
<211>  253
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Aminoacidic sequence of His-VC1-StrepEc

<400>  2

Met Gly Ser Ser His His His His His His Ser Ser Gly Leu Val Pro 
1               5                   10                  15      


Arg Gly Ser His Met Ala Gln Asn Ile Thr Ala Arg Ile Gly Glu Pro 
            20                  25                  30          


Leu Val Leu Lys Cys Lys Gly Ala Pro Lys Lys Pro Pro Gln Arg Leu 
        35                  40                  45              


Glu Trp Lys Leu Asn Thr Gly Arg Thr Glu Ala Trp Lys Val Leu Ser 
    50                  55                  60                  


Pro Gln Gly Gly Gly Pro Trp Asp Ser Val Ala Arg Val Leu Pro Asn 
65                  70                  75                  80  


Gly Ser Leu Phe Leu Pro Ala Val Gly Ile Gln Asp Glu Gly Ile Phe 
                85                  90                  95      


Arg Cys Gln Ala Met Asn Arg Asn Gly Lys Glu Thr Lys Ser Asn Tyr 
            100                 105                 110         


Arg Val Arg Val Tyr Gln Ile Pro Gly Lys Pro Glu Ile Val Asp Ser 
        115                 120                 125             


Ala Ser Glu Leu Thr Ala Gly Val Pro Asn Lys Val Gly Thr Cys Val 
    130                 135                 140                 


Ser Glu Gly Ser Tyr Pro Ala Gly Thr Leu Ser Trp His Leu Asp Gly 
145                 150                 155                 160 


Lys Pro Leu Val Pro Asn Glu Lys Gly Val Ser Val Lys Glu Gln Thr 
                165                 170                 175     


Arg Arg His Pro Glu Thr Gly Leu Phe Thr Leu Gln Ser Glu Leu Met 
            180                 185                 190         


Val Thr Pro Ala Arg Gly Gly Asp Pro Arg Pro Thr Phe Ser Cys Ser 
        195                 200                 205             


Phe Ser Pro Gly Leu Pro Arg His Arg Ala Leu Arg Thr Ala Pro Ile 
    210                 215                 220                 


Gln Pro Arg Val Trp Glu Pro Val Pro Leu Glu Glu Val Gln Leu Val 
225                 230                 235                 240 


Val Glu Gly Ser Ala Trp Arg His Pro Gln Phe Gly Gly 
                245                 250             


<210>  3
<211>  768
<212>  DNA
<213>  artificial sequence

<220>
<223>   Nucleotidic sequence of VC1-StrepPp

<400>  3
atgttctctc caattttgtc cttggaaatt attttagctt tggctacttt gcaatctgtc       60

ttcgctcgag ctcaaaacat cacagcccgg attggcgagc cactggtgct gaagtgtaag      120

ggggccccca agaaaccacc ccagcggctg gaatggaaac tgaacacagg ccggacagaa      180

gcttggaagg tcctgtctcc ccagggagga ggcccctggg acagtgtggc tcgtgtcctt      240

cccaacggct ccctcttcct tccggctgtc gggatccagg atgaggggat tttccggtgc      300

caggcaatga acaggaatgg aaaggagacc aagtccaact accgagtccg tgtctaccag      360

attcctggga agccagaaat tgtagattct gcctctgaac tcacggctgg tgttcccaat      420

aaggtgggga catgtgtgtc agagggaagc taccctgcag ggactcttag ctggcacttg      480

gatgggaagc ccctggtgcc taatgagaag ggagtatctg tgaaggaaca gaccaggaga      540

caccctgaga cagggctctt cacactgcag tcggagctaa tggtgacccc agcccgggga      600

ggagatcccc gtcccacctt ctcctgtagc ttcagcccag gccttccccg acaccgggcc      660

ttgcgcacag cccccatcca gccccgtgtc tgggagcctg tgcctctgga ggaggtccaa      720

ttggtggtgg agggtagcgc ttggcgtcac ccgcagttcg gtggttag                   768


<210>  4
<211>  255
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Aminoacidic sequence of the VC1-StrepPp precursor

<400>  4

Met Phe Ser Pro Ile Leu Ser Leu Glu Ile Ile Leu Ala Leu Ala Thr 
1               5                   10                  15      


Leu Gln Ser Val Phe Ala Arg Ala Gln Asn Ile Thr Ala Arg Ile Gly 
            20                  25                  30          


Glu Pro Leu Val Leu Lys Cys Lys Gly Ala Pro Lys Lys Pro Pro Gln 
        35                  40                  45              


Arg Leu Glu Trp Lys Leu Asn Thr Gly Arg Thr Glu Ala Trp Lys Val 
    50                  55                  60                  


Leu Ser Pro Gln Gly Gly Gly Pro Trp Asp Ser Val Ala Arg Val Leu 
65                  70                  75                  80  


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


Ile Phe Arg Cys Gln Ala Met Asn Arg Asn Gly Lys Glu Thr Lys Ser 
            100                 105                 110         


Asn Tyr Arg Val Arg Val Tyr Gln Ile Pro Gly Lys Pro Glu Ile Val 
        115                 120                 125             


Asp Ser Ala Ser Glu Leu Thr Ala Gly Val Pro Asn Lys Val Gly Thr 
    130                 135                 140                 


Cys Val Ser Glu Gly Ser Tyr Pro Ala Gly Thr Leu Ser Trp His Leu 
145                 150                 155                 160 


Asp Gly Lys Pro Leu Val Pro Asn Glu Lys Gly Val Ser Val Lys Glu 
                165                 170                 175     


Gln Thr Arg Arg His Pro Glu Thr Gly Leu Phe Thr Leu Gln Ser Glu 
            180                 185                 190         


Leu Met Val Thr Pro Ala Arg Gly Gly Asp Pro Arg Pro Thr Phe Ser 
        195                 200                 205             


Cys Ser Phe Ser Pro Gly Leu Pro Arg His Arg Ala Leu Arg Thr Ala 
    210                 215                 220                 


Pro Ile Gln Pro Arg Val Trp Glu Pro Val Pro Leu Glu Glu Val Gln 
225                 230                 235                 240 


Leu Val Val Glu Gly Ser Ala Trp Arg His Pro Gln Phe Gly Gly 
                245                 250                 255 


<210>  5
<211>  705
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotidic sequence of VC1Pp

<400>  5
atgttctctc caattttgtc cttggaaatt attttagctt tggctacttt gcaatctgtc       60

ttcgctcgag ctcaaaacat cacagcccgg attggcgagc cactggtgct gaagtgtaag      120

ggggccccca agaaaccacc ccagcggctg gaatggaaac tgaacacagg ccggacagaa      180

gcttggaagg tcctgtctcc ccagggagga ggcccctggg acagtgtggc tcgtgtcctt      240

cccaacggct ccctcttcct tccggctgtc gggatccagg atgaggggat tttccggtgc      300

caggcaatga acaggaatgg aaaggagacc aagtccaact accgagtccg tgtctaccag      360

attcctggga agccagaaat tgtagattct gcctctgaac tcacggctgg tgttcccaat      420

aaggtgggga catgtgtgtc agagggaagc taccctgcag ggactcttag ctggcacttg      480

gatgggaagc ccctggtgcc taatgagaag ggagtatctg tgaaggaaca gaccaggaga      540

caccctgaga cagggctctt cacactgcag tcggagctaa tggtgacccc agcccgggga      600

ggagatcccc gtcccacctt ctcctgtagc ttcagcccag gccttccccg acaccgggcc      660

ttgcgcacag cccccatcca gccccgtgtc tgggagcctg tgtag                      705


<210>  6
<211>  234
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Aminoacidic sequence of VC1Pp precursor

<400>  6

Met Phe Ser Pro Ile Leu Ser Leu Glu Ile Ile Leu Ala Leu Ala Thr 
1               5                   10                  15      


Leu Gln Ser Val Phe Ala Arg Ala Gln Asn Ile Thr Ala Arg Ile Gly 
            20                  25                  30          


Glu Pro Leu Val Leu Lys Cys Lys Gly Ala Pro Lys Lys Pro Pro Gln 
        35                  40                  45              


Arg Leu Glu Trp Lys Leu Asn Thr Gly Arg Thr Glu Ala Trp Lys Val 
    50                  55                  60                  


Leu Ser Pro Gln Gly Gly Gly Pro Trp Asp Ser Val Ala Arg Val Leu 
65                  70                  75                  80  


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


Ile Phe Arg Cys Gln Ala Met Asn Arg Asn Gly Lys Glu Thr Lys Ser 
            100                 105                 110         


Asn Tyr Arg Val Arg Val Tyr Gln Ile Pro Gly Lys Pro Glu Ile Val 
        115                 120                 125             


Asp Ser Ala Ser Glu Leu Thr Ala Gly Val Pro Asn Lys Val Gly Thr 
    130                 135                 140                 


Cys Val Ser Glu Gly Ser Tyr Pro Ala Gly Thr Leu Ser Trp His Leu 
145                 150                 155                 160 


Asp Gly Lys Pro Leu Val Pro Asn Glu Lys Gly Val Ser Val Lys Glu 
                165                 170                 175     


Gln Thr Arg Arg His Pro Glu Thr Gly Leu Phe Thr Leu Gln Ser Glu 
            180                 185                 190         


Leu Met Val Thr Pro Ala Arg Gly Gly Asp Pro Arg Pro Thr Phe Ser 
        195                 200                 205             


Cys Ser Phe Ser Pro Gly Leu Pro Arg His Arg Ala Leu Arg Thr Ala 
    210                 215                 220                 


Pro Ile Gln Pro Arg Val Trp Glu Pro Val 
225                 230                 


<210>  7
<211>  41
<212>  DNA
<213>  artificial

<220>
<223>  VNdeIFOR for the construction of the recombinant plasmid  pET-15b 
       His-VC1-Strep for the expression in E. coli

<400>  7
agcatattcg actgacatat ggctcaaaac atcacagccc g                           41


<210>  8
<211>  72
<212>  DNA
<213>  Artificial

<220>
<223>  StrepREV for the construction of the recombinant plasmid  pET-15b 
       His-VC1-Strep for the expression in E. coli

<400>  8
atcgtcgggc tcactcgagc taaccaccga actgcgggtg acgccaagcg ctaccctcca       60

ccaccaattg ga                                                           72


<210>  9
<211>  40
<212>  DNA
<213>  artificial

<220>
<223>  VCXhoIFOR for the construction of the recombinant plasmid or the expression of
	an immobilizable variant of VC1 in P. pastoris

<400>  9
agcatattcg actgactcga gctcaaaaca tcacagcccg                             40


<210>  10
<211>  72
<212>  DNA
<213>  artificial

<220>
<223>  StrepREV for the construction of the recombinant plasmid for the expression of
	an immobilizable variant of VC1 in P. pastoris

<400>  10
atcgtcgggc tcactcgagc taaccaccga actgcgggtg acgccaagcg ctaccctcca       60

ccaccaattg ga                                                           72


<210>  11
<211>  40
<212>  DNA
<213>  artificial

<220>
<223>  VCXhoIFOR for the construction of the recombinant plasmid pHIL-S1 
       VC1 for the expression of VC1 in P. pastoris

<400>  11
agcatattcg actgactcga gctcaaaaca tcacagcccg                             40


<210>  12
<211>  40
<212>  DNA
<213>  artificial

<220>
<223>  VC233XhoREV for the construction of the recombinant plasmid pHIL-S1 
       VC1 for the expression of VC1 in P. pastoris


<400>  12
atcgtcgggc tcactcgagc tacacaggct cccagacacg                             40


<210>  13
<211>  235
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Aminoacidic sequence His-VC1-StrepEc of VC1-StrepEc after 
	digestion with thrombin

<400>  13

Gly Ser His Met Ala Gln Asn Ile Thr Ala Arg Ile Gly Glu Pro Leu 
1               5                   10                  15      


Val Leu Lys Cys Lys Gly Ala Pro Lys Lys Pro Pro Gln Arg Leu Glu 
            20                  25                  30          


Trp Lys Leu Asn Thr Gly Arg Thr Glu Ala Trp Lys Val Leu Ser Pro 
        35                  40                  45              


Gln Gly Gly Gly Pro Trp Asp Ser Val Ala Arg Val Leu Pro Asn Gly 
    50                  55                  60                  


Ser Leu Phe Leu Pro Ala Val Gly Ile Gln Asp Glu Gly Ile Phe Arg 
65                  70                  75                  80  


Cys Gln Met Asn Arg Asn Gly Lys Glu Thr Lys Ser Asn Tyr Arg Val 
                85                  90                  95      


Arg Val Tyr Gln Ile Pro Gly Lys Pro Glu Ile Val Asp Ser Ala Ser 
            100                 105                 110         


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


Gly Ser Tyr Pro Ala Gly Thr Leu Ser Trp His Leu Asp Gly Lys Pro 
    130                 135                 140                 


Leu Val Pro Asn Glu Lys Gly Val Ser Val Lys Glu Gln Thr Arg Arg 
145                 150                 155                 160 


His Pro Glu Thr Gly Leu Phe Thr Leu Gln Ser Glu Leu Met Val Thr 
                165                 170                 175     


Pro Ala Arg Gly Gly Asp Pro Arg Pro Thr Phe Ser Cys Ser Phe Ser 
            180                 185                 190         


Pro Gly Leu Pro Arg His Arg Ala Leu Arg Thr Ala Pro Ile Gln Pro 
        195                 200                 205             


Arg Val Trp Glu Pro Val Pro Leu Glu Glu Val Gln Leu Val Val Glu 
    210                 215                 220                 


Gly Ser Ala Trp Arg His Pro Gln Phe Gly Gly 
225                 230                 235 


<210>  14
<211>  233
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Aminoacidic sequence of VC1-StrepPp mature-after removal of 
	the signal sequence

<400>  14

Arg Ala Gln Asn Ile Thr Ala Arg Ile Gly Glu Pro Leu Val Leu Lys 
1               5                   10                  15      


Cys Lys Gly Ala Pro Lys Lys Pro Pro Gln Arg Leu Glu Trp Lys Leu 
            20                  25                  30          


Asn Thr Gly Arg Thr Glu Ala Trp Lys Val Leu Ser Pro Gln Gly Gly 
        35                  40                  45              


Gly Pro Trp Asp Ser Val Ala Arg Val Leu Pro Asn Gly Ser Leu Phe 
    50                  55                  60                  


Leu Pro Ala Val Gly Ile Gln Asp Glu Gly Ile Phe Arg Cys Gln Ala 
65                  70                  75                  80  


Met Asn Arg Asn Gly Lys Glu Thr Lys Ser Asn Tyr Arg Val Arg Val 
                85                  90                  95      


Tyr Gln Ile Pro Gly Lys Pro Glu Ile Val Asp Ser Ala Ser Glu Leu 
            100                 105                 110         


Thr Ala Gly Val Pro Asn Lys Val Gly Thr Cys Val Ser Glu Gly Ser 
        115                 120                 125             


Tyr Pro Ala Gly Thr Leu Ser Trp His Leu Asp Gly Lys Pro Leu Val 
    130                 135                 140                 


Pro Asn Glu Lys Gly Val Ser Val Lys Glu Gln Thr Arg Arg His Pro 
145                 150                 155                 160 


Glu Thr Gly Leu Phe Thr Leu Gln Ser Glu Leu Met Val Thr Pro Ala 
                165                 170                 175     


Arg Gly Gly Asp Pro Arg Pro Thr Phe Ser Cys Ser Phe Ser Pro Gly 
            180                 185                 190         


Leu Pro Arg His Arg Ala Leu Arg Thr Ala Pro Ile Gln Pro Arg Val 
        195                 200                 205             


Trp Glu Pro Val Pro Leu Glu Glu Val Gln Leu Val Val Glu Gly Ser 
    210                 215                 220                 


Ala Trp Arg His Pro Gln Phe Gly Gly 
225                 230             


<210>  15
<211>  212
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Aminoacidic sequence VC1Pp mature-after removal of the signal 
	sequence

<400>  15

Arg Ala Gln Asn Ile Thr Ala Arg Ile Gly Glu Pro Leu Val Leu Lys 
1               5                   10                  15      


Cys Lys Gly Ala Pro Lys Lys Pro Pro Gln Arg Leu Glu Trp Lys Leu 
            20                  25                  30          


Asn Thr Gly Arg Thr Glu Ala Trp Lys Val Leu Ser Pro Gln Gly Gly 
        35                  40                  45              


Gly Pro Trp Asp Ser Val Ala Arg Val Leu Pro Asn Gly Ser Leu Phe 
    50                  55                  60                  


Leu Pro Ala Val Gly Ile Gln Asp Glu Gly Ile Phe Arg Cys Gln Ala 
65                  70                  75                  80  


Met Asn Arg Asn Gly Lys Glu Thr Lys Ser Asn Tyr Arg Val Arg Val 
                85                  90                  95      


Tyr Gln Ile Pro Gly Lys Pro Glu Ile Val Asp Ser Ala Ser Glu Leu 
            100                 105                 110         


Thr Ala Gly Val Pro Asn Lys Val Gly Thr Cys Val Ser Glu Gly Ser 
        115                 120                 125             


Tyr Pro Ala Gly Thr Leu Ser Trp His Leu Asp Gly Lys Pro Leu Val 
    130                 135                 140                 


Pro Asn Glu Lys Gly Val Ser Val Lys Glu Gln Thr Arg Arg His Pro 
145                 150                 155                 160 


Glu Thr Gly Leu Phe Thr Leu Gln Ser Glu Leu Met Val Thr Pro Ala 
                165                 170                 175     


Arg Gly Gly Asp Pro Arg Pro Thr Phe Ser Cys Ser Phe Ser Pro Gly 
            180                 185                 190         


Leu Pro Arg His Arg Ala Leu Arg Thr Ala Pro Ile Gln Pro Arg Val 
        195                 200                 205             


Trp Glu Pro Val 
    210         


<210>  16
<211>  735
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotidic sequence of VC1-HisPp

<400>  16
atgttctctc caattttgtc cttggaaatt attttagctt tggctacttt gcaatctgtc       60

ttcgctcgag ctcaaaacat cacagcccgg attggcgagc cactggtgct gaagtgtaag      120

ggggccccca agaaaccacc ccagcggctg gaatggaaac tgaacacagg ccggacagaa      180

gcttggaagg tcctgtctcc ccagggagga ggcccctggg acagtgtggc tcgtgtcctt      240

cccaacggct ccctcttcct tccggctgtc gggatccagg atgaggggat tttccggtgc      300

caggcaatga acaggaatgg aaaggagacc aagtccaact accgagtccg tgtctaccag      360

attcctggga agccagaaat tgtagattct gcctctgaac tcacggctgg tgttcccaat      420

aaggtgggga catgtgtgtc agagggaagc taccctgcag ggactcttag ctggcacttg      480

gatgggaagc ccctggtgcc taatgagaag ggagtatctg tgaaggaaca gaccaggaga      540

caccctgaga cagggctctt cacactgcag tcggagctaa tggtgacccc agcccgggga      600

ggagatcccc gtcccacctt ctcctgtagc ttcagcccag gccttccccg acaccgggcc      660

ttgcgcacag cccccatcca gccccgtgtc tgggagcctg tgggcagcca tcaccatcac      720

catcactagc tcgag                                                       735


<210>  17
<211>  242
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Aminoacidic sequence of the VC1-HisPp precursor 

<400>  17

Met Phe Ser Pro Ile Leu Ser Leu Glu Ile Ile Leu Ala Leu Ala Thr 
1               5                   10                  15      


Leu Gln Ser Val Phe Ala Arg Ala Gln Asn Ile Thr Ala Arg Ile Gly 
            20                  25                  30          


Glu Pro Leu Val Leu Lys Cys Lys Gly Ala Pro Lys Lys Pro Pro Gln 
        35                  40                  45              


Arg Leu Glu Trp Lys Leu Asn Thr Gly Arg Thr Glu Ala Trp Lys Val 
    50                  55                  60                  


Leu Ser Pro Gln Gly Gly Gly Pro Trp Asp Ser Val Ala Arg Val Leu 
65                  70                  75                  80  


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


Ile Phe Arg Cys Gln Ala Met Asn Arg Asn Gly Lys Glu Thr Lys Ser 
            100                 105                 110         


Asn Tyr Arg Val Arg Val Tyr Gln Ile Pro Gly Lys Pro Glu Ile Val 
        115                 120                 125             


Asp Ser Ala Ser Glu Leu Thr Ala Gly Val Pro Asn Lys Val Gly Thr 
    130                 135                 140                 


Cys Val Ser Glu Gly Ser Tyr Pro Ala Gly Thr Leu Ser Trp His Leu 
145                 150                 155                 160 


Asp Gly Lys Pro Leu Val Pro Asn Glu Lys Gly Val Ser Val Lys Glu 
                165                 170                 175     


Gln Thr Arg Arg His Pro Glu Thr Gly Leu Phe Thr Leu Gln Ser Glu 
            180                 185                 190         


Leu Met Val Thr Pro Ala Arg Gly Gly Asp Pro Arg Pro Thr Phe Ser 
        195                 200                 205             


Cys Ser Phe Ser Pro Gly Leu Pro Arg His Arg Ala Leu Arg Thr Ala 
    210                 215                 220                 


Pro Ile Gln Pro Arg Val Trp Glu Pro Val Gly Ser His His His His 
225                 230                 235                 240 


His His 
        


<210>  18
<211>  220
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Aminoacidic sequence of VC1-HisPp mature-after removal of the 
	signal sequence

<400>  18

Arg Ala Gln Asn Ile Thr Ala Arg Ile Gly Glu Pro Leu Val Leu Lys 
1               5                   10                  15      


Cys Lys Gly Ala Pro Lys Lys Pro Pro Gln Arg Leu Glu Trp Lys Leu 
            20                  25                  30          


Asn Thr Gly Arg Thr Glu Ala Trp Lys Val Leu Ser Pro Gln Gly Gly 
        35                  40                  45              


Gly Pro Trp Asp Ser Val Ala Arg Val Leu Pro Asn Gly Ser Leu Phe 
    50                  55                  60                  


Leu Pro Ala Val Gly Ile Gln Asp Glu Gly Ile Phe Arg Cys Gln Ala 
65                  70                  75                  80  


Met Asn Arg Asn Gly Lys Glu Thr Lys Ser Asn Tyr Arg Val Arg Val 
                85                  90                  95      


Tyr Gln Ile Pro Gly Lys Pro Glu Ile Val Asp Ser Ala Ser Glu Leu 
            100                 105                 110         


Thr Ala Gly Val Pro Asn Lys Val Gly Thr Cys Val Ser Glu Gly Ser 
        115                 120                 125             


Tyr Pro Ala Gly Thr Leu Ser Trp His Leu Asp Gly Lys Pro Leu Val 
    130                 135                 140                 


Pro Asn Glu Lys Gly Val Ser Val Lys Glu Gln Thr Arg Arg His Pro 
145                 150                 155                 160 


Glu Thr Gly Leu Phe Thr Leu Gln Ser Glu Leu Met Val Thr Pro Ala 
                165                 170                 175     


Arg Gly Gly Asp Pro Arg Pro Thr Phe Ser Cys Ser Phe Ser Pro Gly 
            180                 185                 190         


Leu Pro Arg His Arg Ala Leu Arg Thr Ala Pro Ile Gln Pro Arg Val 
        195                 200                 205             


Trp Glu Pro Val Gly Ser His His His His His His 
    210                 215                 220 


<210>  19
<211>  795
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotidic sequence of VC1-His-StrepPp

<400>  19
atgttctctc caattttgtc cttggaaatt attttagctt tggctacttt gcaatctgtc       60

ttcgctcgag ctcaaaacat cacagcccgg attggcgagc cactggtgct gaagtgtaag      120

ggggccccca agaaaccacc ccagcggctg gaatggaaac tgaacacagg ccggacagaa      180

gcttggaagg tcctgtctcc ccagggagga ggcccctggg acagtgtggc tcgtgtcctt      240

cccaacggct ccctcttcct tccggctgtc gggatccagg atgaggggat tttccggtgc      300

caggcaatga acaggaatgg aaaggagacc aagtccaact accgagtccg tgtctaccag      360

attcctggga agccagaaat tgtagattct gcctctgaac tcacggctgg tgttcccaat      420

aaggtgggga catgtgtgtc agagggaagc taccctgcag ggactcttag ctggcacttg      480

gatgggaagc ccctggtgcc taatgagaag ggagtatctg tgaaggaaca gaccaggaga      540

caccctgaga cagggctctt cacactgcag tcggagctaa tggtgacccc agcccgggga      600

ggagatcccc gtcccacctt ctcctgtagc ttcagcccag gccttccccg acaccgggcc      660

ttgcgcacag cccccatcca gccccgtgtc tgggagcctg tgagcgccca tcaccatcac      720

catcacggtg gtggtagcgg tggtggtagc ggtggtagcg cttggcgtca cccgcagttc      780

ggtggttagc tcgag                                                       795


<210>  20
<211>  262
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Aminoacidic sequence of the VC1-His-StrepPp precursor

<400>  20

Met Phe Ser Pro Ile Leu Ser Leu Glu Ile Ile Leu Ala Leu Ala Thr 
1               5                   10                  15      


Leu Gln Ser Val Phe Ala Arg Ala Gln Asn Ile Thr Ala Arg Ile Gly 
            20                  25                  30          


Glu Pro Leu Val Leu Lys Cys Lys Gly Ala Pro Lys Lys Pro Pro Gln 
        35                  40                  45              


Arg Leu Glu Trp Lys Leu Asn Thr Gly Arg Thr Glu Ala Trp Lys Val 
    50                  55                  60                  


Leu Ser Pro Gln Gly Gly Gly Pro Trp Asp Ser Val Ala Arg Val Leu 
65                  70                  75                  80  


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


Ile Phe Arg Cys Gln Ala Met Asn Arg Asn Gly Lys Glu Thr Lys Ser 
            100                 105                 110         


Asn Tyr Arg Val Arg Val Tyr Gln Ile Pro Gly Lys Pro Glu Ile Val 
        115                 120                 125             


Asp Ser Ala Ser Glu Leu Thr Ala Gly Val Pro Asn Lys Val Gly Thr 
    130                 135                 140                 


Cys Val Ser Glu Gly Ser Tyr Pro Ala Gly Thr Leu Ser Trp His Leu 
145                 150                 155                 160 


Asp Gly Lys Pro Leu Val Pro Asn Glu Lys Gly Val Ser Val Lys Glu 
                165                 170                 175     


Gln Thr Arg Arg His Pro Glu Thr Gly Leu Phe Thr Leu Gln Ser Glu 
            180                 185                 190         


Leu Met Val Thr Pro Ala Arg Gly Gly Asp Pro Arg Pro Thr Phe Ser 
        195                 200                 205             


Cys Ser Phe Ser Pro Gly Leu Pro Arg His Arg Ala Leu Arg Thr Ala 
    210                 215                 220                 


Pro Ile Gln Pro Arg Val Trp Glu Pro Val Ser Ala His His His His 
225                 230                 235                 240 


His His Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Ser Ala Trp Arg 
                245                 250                 255     


His Pro Gln Phe Gly Gly 
            260         


<210>  21
<211>  240
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Aminoacidic sequence of VC1-His-StrepPp mature-after removal 
	of the signal sequence 

<400>  21

Arg Ala Gln Asn Ile Thr Ala Arg Ile Gly Glu Pro Leu Val Leu Lys 
1               5                   10                  15      


Cys Lys Gly Ala Pro Lys Lys Pro Pro Gln Arg Leu Glu Trp Lys Leu 
            20                  25                  30          


Asn Thr Gly Arg Thr Glu Ala Trp Lys Val Leu Ser Pro Gln Gly Gly 
        35                  40                  45              


Gly Pro Trp Asp Ser Val Ala Arg Val Leu Pro Asn Gly Ser Leu Phe 
    50                  55                  60                  


Leu Pro Ala Val Gly Ile Gln Asp Glu Gly Ile Phe Arg Cys Gln Ala 
65                  70                  75                  80  


Met Asn Arg Asn Gly Lys Glu Thr Lys Ser Asn Tyr Arg Val Arg Val 
                85                  90                  95      


Tyr Gln Ile Pro Gly Lys Pro Glu Ile Val Asp Ser Ala Ser Glu Leu 
            100                 105                 110         


Thr Ala Gly Val Pro Asn Lys Val Gly Thr Cys Val Ser Glu Gly Ser 
        115                 120                 125             


Tyr Pro Ala Gly Thr Leu Ser Trp His Leu Asp Gly Lys Pro Leu Val 
    130                 135                 140                 


Pro Asn Glu Lys Gly Val Ser Val Lys Glu Gln Thr Arg Arg His Pro 
145                 150                 155                 160 


Glu Thr Gly Leu Phe Thr Leu Gln Ser Glu Leu Met Val Thr Pro Ala 
                165                 170                 175     


Arg Gly Gly Asp Pro Arg Pro Thr Phe Ser Cys Ser Phe Ser Pro Gly 
            180                 185                 190         


Leu Pro Arg His Arg Ala Leu Arg Thr Ala Pro Ile Gln Pro Arg Val 
        195                 200                 205             


Trp Glu Pro Val Ser Ala His His His His His His Gly Gly Gly Ser 
    210                 215                 220                 


Gly Gly Gly Ser Gly Gly Ser Ala Trp Arg His Pro Gln Phe Gly Gly 
225                 230                 235                 240 


<210>  22
<211>  64
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  VC1-233HisREV for the construction of the recombinant plasmid for 
	the expression of the immobilizzable variant VC1-HisPp in P. pastoris

<400>  22
atcgtcgggc tcactcgagc tagtgatggt gatggtgatg gctgcccaca ggctcccaga       60

cacg                                                                    64


<210>  23
<211>  124
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  VC1-233His-StrepREV for the construction of the recombinant plasmid for 
	the expression of the immobilizzable VC1-His-StrepPp variant in P. pastoris

<400>  23
atcgtcgggc tcactcgagc taaccaccga actgcgggtg acgccaagcg ctaccaccgc       60

taccaccacc gctaccacca ccgtgatggt gatggtgatg ggcgctcaca ggctcccaga      120

cacg                                                                   124


<210>  24
<211>  8
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Strep tag

<400>  24

Trp Arg His Pro Gln Phe Gly Gly 
1               5               

