                         SEQUENCE LISTING

<110>  Techinical University of Denmark
 
<120>  A mutant sialidase having trans-sialidase activity for use in 
       production of sialylated glycans

<130>  P1419EP00

<160>  40    

<170>  PatentIn version 3.5

<210>  1
<211>  641
<212>  PRT
<213>  Trypanosoma rangeli


<220>
<221>  PEPTIDE
<222>  (1)..(641)
<223>  T. rangeli mutant sialidase (PBD 1WCS)

<400>  1

Ala Ala Ser Leu Ala Pro Gly Ser Ser Arg Val Glu Leu Phe Lys Arg 
1               5                   10                  15      


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


Arg Val Val His Ser Phe Arg Ile Pro Thr Ile Val Asn Val Asp Gly 
        35                  40                  45              


Val Met Val Ala Ile Ala Asp Ala Arg Tyr Glu Thr Ser Phe Asp Asn 
    50                  55                  60                  


Ser Phe Ile Glu Thr Ala Val Lys Tyr Ser Val Asp Asp Gly Ala Thr 
65                  70                  75                  80  


Trp Asn Thr Gln Ile Ala Ile Lys Asn Ser Arg Ala Ser Ser Val Ser 
                85                  90                  95      


Arg Val Val Asp Pro Thr Val Ile Val Lys Gly Asn Lys Leu Tyr Ile 
            100                 105                 110         


Leu Val Gly Ser Phe Asn Lys Thr Arg Asn Tyr Trp Thr Gln His Arg 
        115                 120                 125             


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


Ser Ala Ala Asn Gly Lys Thr Thr Ala Thr Ile Ser Trp Gly Lys Pro 
145                 150                 155                 160 


Val Ser Leu Lys Pro Leu Phe Pro Ala Glu Phe Asp Gly Ile Leu Thr 
                165                 170                 175     


Lys Glu Phe Val Gly Gly Val Gly Ala Ala Ile Val Ala Ser Asn Gly 
            180                 185                 190         


Asn Leu Val Tyr Pro Val Gln Ile Ala Asp Met Gly Gly Arg Val Phe 
        195                 200                 205             


Thr Lys Ile Met Tyr Ser Glu Asp Asp Gly Asn Thr Trp Lys Phe Ala 
    210                 215                 220                 


Glu Gly Arg Ser Lys Phe Gly Cys Ser Glu Pro Ala Val Leu Glu Trp 
225                 230                 235                 240 


Glu Gly Lys Leu Ile Ile Asn Asn Arg Val Asp Tyr Asn Arg Arg Leu 
                245                 250                 255     


Val Tyr Glu Ser Ser Asp Met Gly Lys Thr Trp Val Glu Ala Leu Gly 
            260                 265                 270         


Thr Leu Ser His Val Trp Thr Asn Ser Pro Thr Ser Asn Gln Pro Asp 
        275                 280                 285             


Cys Gln Ser Ser Phe Val Ala Val Thr Ile Glu Gly Lys Arg Val Met 
    290                 295                 300                 


Leu Phe Thr His Pro Leu Asn Leu Lys Gly Arg Trp Met Arg Asp Arg 
305                 310                 315                 320 


Leu His Leu Trp Met Thr Asp Asn Gln Arg Ile Phe Asp Val Gly Gln 
                325                 330                 335     


Ile Ser Ile Gly Asp Glu Asn Ser Gly Tyr Ser Ser Val Leu Tyr Lys 
            340                 345                 350         


Asp Asp Lys Leu Tyr Ser Leu His Glu Ile Asn Thr Asn Asp Val Tyr 
        355                 360                 365             


Ser Leu Val Phe Val Arg Leu Ile Gly Glu Leu Gln Leu Met Lys Ser 
    370                 375                 380                 


Val Val Arg Thr Trp Lys Glu Glu Asp Asn His Leu Ala Ser Ile Cys 
385                 390                 395                 400 


Thr Pro Val Val Pro Ala Thr Pro Pro Ser Lys Gly Ala Cys Gly Ala 
                405                 410                 415     


Ala Val Pro Thr Ala Gly Leu Val Gly Phe Leu Ser His Ser Ala Asn 
            420                 425                 430         


Gly Ser Val Trp Glu Asp Val Tyr Arg Cys Val Asp Ala Asn Val Ala 
        435                 440                 445             


Asn Ala Glu Arg Val Pro Asn Gly Leu Lys Phe Asn Gly Val Gly Gly 
    450                 455                 460                 


Gly Ala Val Trp Pro Val Ala Arg Gln Gly Gln Thr Arg Arg Tyr Gln 
465                 470                 475                 480 


Phe Ala Asn Tyr Arg Phe Thr Leu Val Ala Thr Val Thr Ile Asp Glu 
                485                 490                 495     


Leu Pro Lys Gly Thr Ser Pro Leu Leu Gly Ala Gly Leu Glu Gly Pro 
            500                 505                 510         


Gly Asp Ala Lys Leu Leu Gly Leu Ser Tyr Asp Lys Asn Arg Gln Trp 
        515                 520                 525             


Arg Pro Leu Tyr Gly Ala Ala Pro Ala Ser Pro Thr Gly Ser Trp Glu 
    530                 535                 540                 


Leu His Lys Lys Tyr His Val Val Leu Thr Met Ala Asp Arg Gln Gly 
545                 550                 555                 560 


Ser Val Tyr Val Asp Gly Gln Pro Leu Ala Gly Ser Gly Asn Thr Val 
                565                 570                 575     


Val Arg Gly Ala Thr Leu Pro Asp Ile Ser His Phe Tyr Ile Gly Gly 
            580                 585                 590         


Pro Arg Ser Lys Gly Ala Pro Thr Asp Ser Arg Val Thr Val Thr Asn 
        595                 600                 605             


Val Val Leu Tyr Asn Arg Arg Leu Asn Ser Ser Glu Ile Arg Thr Leu 
    610                 615                 620                 


Phe Leu Ser Gln Asp Met Ile Gly Thr Asp Gly Gly Ala Gly Thr Ala 
625                 630                 635                 640 


Ala 
    


<210>  2
<211>  638
<212>  PRT
<213>  Trypanosoma rangeli


<220>
<221>  mat_peptide
<222>  (1)..(638)

<220>
<221>  PEPTIDE
<222>  (1)..(372)
<223>  Mutant Tr6 T. rangeli trans-sialiadase - N-terminal region and 
       catalytic peptide

<220>
<221>  PEPTIDE
<222>  (28)..(372)
<223>  Mutant Tr6 T. rangeli trans-sialiadase - catalytic peptide

<220>
<221>  PEPTIDE
<222>  (373)..(404)
<223>  Mutant Tr6 T. rangeli trans-sialiadase - linker peptide

<220>
<221>  PEPTIDE
<222>  (405)..(638)
<223>  Mutant Tr6 T. rangeli trans-sialiadase - carbohydrate-binding 
       peptide

<400>  2

Leu Ala Pro Gly Ser Ser Arg Val Glu Leu Phe Lys Arg Lys Asn Ser 
1               5                   10                  15      


Thr Val Pro Phe Glu Glu Ser Asn Gly Thr Ile Arg Glu Arg Val Val 
            20                  25                  30          


His Ser Phe Arg Leu Pro Thr Ile Val Asn Val Asp Gly Val Met Val 
        35                  40                  45              


Ala Ile Ala Asp Ala Arg Tyr Glu Thr Ser Phe Asp Asn Ser Phe Ile 
    50                  55                  60                  


Glu Thr Ala Val Lys Tyr Ser Val Asp Asp Gly Ala Thr Trp Asn Thr 
65                  70                  75                  80  


Gln Ile Ala Ile Lys Asn Ser Arg Ala Ser Ser Val Ser Arg Val Val 
                85                  90                  95      


Asp Pro Thr Val Ile Val Lys Gly Asn Lys Leu Tyr Ile Leu Val Gly 
            100                 105                 110         


Ser Phe Asn Lys Thr Arg Asn Tyr Trp Thr Gln His Arg Asp Gly Ser 
        115                 120                 125             


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


Asn Gly Lys Thr Thr Ala Thr Ile Ser Trp Gly Lys Pro Val Ser Leu 
145                 150                 155                 160 


Lys Pro Leu Phe Pro Ala Glu Phe Asp Gly Ile Leu Thr Lys Glu Phe 
                165                 170                 175     


Val Gly Gly Val Gly Ala Ala Ile Val Ala Ser Asn Gly Asn Leu Val 
            180                 185                 190         


Tyr Pro Val Gln Ile Ala Asp Met Gly Gly Arg Val Phe Thr Lys Ile 
        195                 200                 205             


Met Tyr Ser Glu Asp Asp Gly Asn Thr Trp Lys Phe Ala Glu Gly Arg 
    210                 215                 220                 


Ser Lys Phe Gly Cys Ser Glu Pro Ala Val Leu Glu Trp Glu Gly Lys 
225                 230                 235                 240 


Leu Ile Ile Asn Asn Arg Val Asp Tyr Asn Arg Arg Leu Val Tyr Glu 
                245                 250                 255     


Ser Ser Asp Met Gly Lys Thr Trp Val Glu Ala Leu Gly Thr Leu Ser 
            260                 265                 270         


His Val Trp Thr Asn Ser Pro Thr Ser Asn Gln Pro Asp Cys Gln Ser 
        275                 280                 285             


Ser Phe Val Ala Val Thr Ile Glu Gly Lys Arg Val Met Leu Phe Thr 
    290                 295                 300                 


His Pro Leu Asn Leu Lys Gly Arg Trp Met Arg Asp Arg Leu His Leu 
305                 310                 315                 320 


Trp Met Thr Asp Asn Gln Arg Ile Phe Asp Val Gly Gln Ile Ser Ile 
                325                 330                 335     


Gly Asp Glu Asn Ser Gly Tyr Ser Ser Val Leu Tyr Lys Asp Asp Lys 
            340                 345                 350         


Leu Tyr Ser Leu His Glu Ile Asn Thr Asn Asp Val Tyr Ser Leu Val 
        355                 360                 365             


Phe Val Arg Leu Ile Gly Glu Leu Gln Leu Met Lys Ser Val Val Arg 
    370                 375                 380                 


Thr Trp Lys Glu Glu Asp Asn His Leu Ala Ser Ile Cys Thr Pro Val 
385                 390                 395                 400 


Val Pro Ala Thr Pro Pro Ser Lys Gly Ala Cys Gly Ala Ala Val Pro 
                405                 410                 415     


Thr Ala Gly Leu Val Gly Phe Leu Ser His Ser Ala Asn Gly Ser Val 
            420                 425                 430         


Trp Glu Asp Val Tyr Arg Cys Val Asp Ala Asn Val Ala Asn Ala Glu 
        435                 440                 445             


Arg Val Pro Asn Gly Leu Lys Phe Asn Gly Val Gly Gly Gly Ala Val 
    450                 455                 460                 


Trp Pro Val Ala Arg Gln Gly Gln Thr Arg Arg Tyr Gln Phe Ala Asn 
465                 470                 475                 480 


Tyr Arg Phe Thr Leu Val Ala Thr Val Thr Ile Asp Glu Leu Pro Lys 
                485                 490                 495     


Gly Thr Ser Pro Leu Leu Gly Ala Gly Leu Glu Gly Pro Gly Asp Ala 
            500                 505                 510         


Lys Leu Leu Gly Leu Ser Tyr Asp Lys Asn Arg Gln Trp Arg Pro Leu 
        515                 520                 525             


Tyr Gly Ala Ala Pro Ala Ser Pro Thr Gly Ser Trp Glu Leu His Lys 
    530                 535                 540                 


Lys Tyr His Val Val Leu Thr Met Ala Asp Arg Gln Gly Ser Val Tyr 
545                 550                 555                 560 


Val Asp Gly Gln Pro Leu Ala Gly Ser Gly Asn Thr Val Val Arg Gly 
                565                 570                 575     


Ala Thr Leu Pro Asp Ile Ser His Phe Tyr Ile Gly Gly Pro Arg Ser 
            580                 585                 590         


Lys Gly Ala Pro Thr Asp Ser Arg Val Thr Val Thr Asn Val Val Leu 
        595                 600                 605             


Tyr Asn Arg Arg Leu Asn Ser Ser Glu Ile Arg Thr Leu Phe Leu Ser 
    610                 615                 620                 


Gln Asp Met Ile Gly Thr Asp Gly Gly Ala Gly Thr Ala Ala 
625                 630                 635             


<210>  3
<211>  1914
<212>  DNA
<213>  Trypanosoma rangeli


<220>
<221>  CDS
<222>  (1)..(1914)
<223>  CDS for Tr13 mutant R. rangeli trans-sialidase comprising: 
       N-terminal region to catalytic domain peptide: aa residues 1-81; 
       Catalytic domain peptide: aa residues 81-1116; Linker peptide: aa
       residues 1117-1212; CBD: aa residues 1213-1914.

<400>  3
ttg gct ccc gga tca tct cgt gtg gaa tta ttt aaa aga aaa aac tcc         48
Leu Ala Pro Gly Ser Ser Arg Val Glu Leu Phe Lys Arg Lys Asn Ser           
1               5                   10                  15                

acc gtg cca ttt gag gag tca aac ggt act ata cga gaa cgt gtg gtt         96
Thr Val Pro Phe Glu Glu Ser Asn Gly Thr Ile Arg Glu Arg Val Val           
            20                  25                  30                    

cat tca ttt aga tta cca act atc gtt aac gta gat gga gtc atg gtt        144
His Ser Phe Arg Leu Pro Thr Ile Val Asn Val Asp Gly Val Met Val           
        35                  40                  45                        

gcc att gct gat gcc aga tat gag aca tca ttc gac aac tcc ttt atc        192
Ala Ile Ala Asp Ala Arg Tyr Glu Thr Ser Phe Asp Asn Ser Phe Ile           
    50                  55                  60                            

gaa act gct gtt aaa tac agt gtt gac gat ggt gct acg tgg aat aca        240
Glu Thr Ala Val Lys Tyr Ser Val Asp Asp Gly Ala Thr Trp Asn Thr           
65                  70                  75                  80            

caa att gca atc aaa aat tct cgt gca tca tca gtt tca agg gtt gtc        288
Gln Ile Ala Ile Lys Asn Ser Arg Ala Ser Ser Val Ser Arg Val Val           
                85                  90                  95                

gat cct acg gtc ata gta aag gga aat aag ttg tat atc ctg gtt gga        336
Asp Pro Thr Val Ile Val Lys Gly Asn Lys Leu Tyr Ile Leu Val Gly           
            100                 105                 110                   

tcc ttt aac aag aca agg aac tat tgg acc cag cac aga gat gga tct        384
Ser Phe Asn Lys Thr Arg Asn Tyr Trp Thr Gln His Arg Asp Gly Ser           
        115                 120                 125                       

gac tgg gaa cca ttg ttg gtg gtt gga gag gtt acg aag tct gct gct        432
Asp Trp Glu Pro Leu Leu Val Val Gly Glu Val Thr Lys Ser Ala Ala           
    130                 135                 140                           

aac ggt aaa aca act gca act att tca tgg ggg aaa cct gtc tcc ctt        480
Asn Gly Lys Thr Thr Ala Thr Ile Ser Trp Gly Lys Pro Val Ser Leu           
145                 150                 155                 160           

aag cct ttg ttc cct gca gag ttc gac ggc ata ctt act aag gaa ttc        528
Lys Pro Leu Phe Pro Ala Glu Phe Asp Gly Ile Leu Thr Lys Glu Phe           
                165                 170                 175               

gta ggt gga gta ggc gcc gcc atc gtg gca agt aat ggt aat ttg gta        576
Val Gly Gly Val Gly Ala Ala Ile Val Ala Ser Asn Gly Asn Leu Val           
            180                 185                 190                   

tac cct gtg caa gta act aat aag aag aag caa gta ttt aca aaa att        624
Tyr Pro Val Gln Val Thr Asn Lys Lys Lys Gln Val Phe Thr Lys Ile           
        195                 200                 205                       

atg tat tcc gag gat gat ggt aac act tgg aag ttc gcc gaa gga agg        672
Met Tyr Ser Glu Asp Asp Gly Asn Thr Trp Lys Phe Ala Glu Gly Arg           
    210                 215                 220                           

tct aag ttc ggt tgc tca gaa cca gca gtt ttg gaa tgg gaa gga aag        720
Ser Lys Phe Gly Cys Ser Glu Pro Ala Val Leu Glu Trp Glu Gly Lys           
225                 230                 235                 240           

cta atc att aat aac cga gtc gat tac aat aga cgt ctg gtg tac gaa        768
Leu Ile Ile Asn Asn Arg Val Asp Tyr Asn Arg Arg Leu Val Tyr Glu           
                245                 250                 255               

tcc agt gac atg ggc aaa aca tgg gta gag gct ctt ggt act ctg tcc        816
Ser Ser Asp Met Gly Lys Thr Trp Val Glu Ala Leu Gly Thr Leu Ser           
            260                 265                 270                   

cac gtc tgg acg aac agt cca act tcc aat caa ccc gat tgt cag agt        864
His Val Trp Thr Asn Ser Pro Thr Ser Asn Gln Pro Asp Cys Gln Ser           
        275                 280                 285                       

tca ttc gtt gca gtt act atc gaa ggt aaa cga gtg atg ttg ttt act        912
Ser Phe Val Ala Val Thr Ile Glu Gly Lys Arg Val Met Leu Phe Thr           
    290                 295                 300                           

cat cca cta aat ttg aag ggt aga tgg atg agg gat aga ctt cat ctg        960
His Pro Leu Asn Leu Lys Gly Arg Trp Met Arg Asp Arg Leu His Leu           
305                 310                 315                 320           

tgg atg acc gat aat cag aga atc ttt gat gtt ggc caa att tcc att       1008
Trp Met Thr Asp Asn Gln Arg Ile Phe Asp Val Gly Gln Ile Ser Ile           
                325                 330                 335               

ggt gat gaa aac agt ggt tac tct tcc gtc cta tac aag gac gat aaa       1056
Gly Asp Glu Asn Ser Gly Tyr Ser Ser Val Leu Tyr Lys Asp Asp Lys           
            340                 345                 350                   

tta tat tcc cta cat gag att aat act aat gat gtt tat tct ctt gtt       1104
Leu Tyr Ser Leu His Glu Ile Asn Thr Asn Asp Val Tyr Ser Leu Val           
        355                 360                 365                       

ttt gtc cga ttg att ggt gag ctg cag tta atg aaa agt gtg gtt cgt       1152
Phe Val Arg Leu Ile Gly Glu Leu Gln Leu Met Lys Ser Val Val Arg           
    370                 375                 380                           

acc tgg aag gaa gag gac aat cat ttg gct tca ata tgt act cca gtc       1200
Thr Trp Lys Glu Glu Asp Asn His Leu Ala Ser Ile Cys Thr Pro Val           
385                 390                 395                 400           

gta cca gca acc cca cca agt aaa gga gcc tgc ggt gcc gct gta cct       1248
Val Pro Ala Thr Pro Pro Ser Lys Gly Ala Cys Gly Ala Ala Val Pro           
                405                 410                 415               

aca gct ggt tta gtt ggc ttc tta tct cac tca gct aat gga tcc gtt       1296
Thr Ala Gly Leu Val Gly Phe Leu Ser His Ser Ala Asn Gly Ser Val           
            420                 425                 430                   

tgg gag gac gta tat aga tgt gtc gat gct aac gtc gcc aac gct gag       1344
Trp Glu Asp Val Tyr Arg Cys Val Asp Ala Asn Val Ala Asn Ala Glu           
        435                 440                 445                       

aga gtt cct aac ggc ctt aag ttt aat ggg gtt ggt ggg ggc gct gtc       1392
Arg Val Pro Asn Gly Leu Lys Phe Asn Gly Val Gly Gly Gly Ala Val           
    450                 455                 460                           

tgg cca gtc gcc agg cag gga caa acc cga agg tac caa ttc gca aac       1440
Trp Pro Val Ala Arg Gln Gly Gln Thr Arg Arg Tyr Gln Phe Ala Asn           
465                 470                 475                 480           

tac aga ttt acc tta gtc gcc acc gtt acg att gac gaa ttg ccc aaa       1488
Tyr Arg Phe Thr Leu Val Ala Thr Val Thr Ile Asp Glu Leu Pro Lys           
                485                 490                 495               

ggt acc tct ccc ctt ctt ggt gcc ggg tta gaa ggt cca ggc gac gct       1536
Gly Thr Ser Pro Leu Leu Gly Ala Gly Leu Glu Gly Pro Gly Asp Ala           
            500                 505                 510                   

aaa ttg cta ggt tta tct tac gac aag aac cgt caa tgg cga ccc ttg       1584
Lys Leu Leu Gly Leu Ser Tyr Asp Lys Asn Arg Gln Trp Arg Pro Leu           
        515                 520                 525                       

tac gga gca gcc cct gct tct cct aca gga tct tgg gag cta cac aag       1632
Tyr Gly Ala Ala Pro Ala Ser Pro Thr Gly Ser Trp Glu Leu His Lys           
    530                 535                 540                           

aag tac cat gta gtc ctg acc atg gct gac aga cag ggg agt gtt tat       1680
Lys Tyr His Val Val Leu Thr Met Ala Asp Arg Gln Gly Ser Val Tyr           
545                 550                 555                 560           

gtt gat ggg caa cct ctt gcc gga tca ggc aat acc gtg gtt aga gga       1728
Val Asp Gly Gln Pro Leu Ala Gly Ser Gly Asn Thr Val Val Arg Gly           
                565                 570                 575               

gct act ttg cca gac atc tct cac ttc tac att ggt gga ccc aga tct       1776
Ala Thr Leu Pro Asp Ile Ser His Phe Tyr Ile Gly Gly Pro Arg Ser           
            580                 585                 590                   

aag gga gcc cct aca gat agt aga gtt aca gtg act aac gtg gtt tta       1824
Lys Gly Ala Pro Thr Asp Ser Arg Val Thr Val Thr Asn Val Val Leu           
        595                 600                 605                       

tat aat cgt aga ctg aac tct agt gag atc cgt aca ctt ttt ttg tct       1872
Tyr Asn Arg Arg Leu Asn Ser Ser Glu Ile Arg Thr Leu Phe Leu Ser           
    610                 615                 620                           

caa gac atg ata ggt acc gat ggt gga gct ggt aca gca gca               1914
Gln Asp Met Ile Gly Thr Asp Gly Gly Ala Gly Thr Ala Ala                   
625                 630                 635                               


<210>  4
<211>  638
<212>  PRT
<213>  Trypanosoma rangeli

<400>  4

Leu Ala Pro Gly Ser Ser Arg Val Glu Leu Phe Lys Arg Lys Asn Ser 
1               5                   10                  15      


Thr Val Pro Phe Glu Glu Ser Asn Gly Thr Ile Arg Glu Arg Val Val 
            20                  25                  30          


His Ser Phe Arg Leu Pro Thr Ile Val Asn Val Asp Gly Val Met Val 
        35                  40                  45              


Ala Ile Ala Asp Ala Arg Tyr Glu Thr Ser Phe Asp Asn Ser Phe Ile 
    50                  55                  60                  


Glu Thr Ala Val Lys Tyr Ser Val Asp Asp Gly Ala Thr Trp Asn Thr 
65                  70                  75                  80  


Gln Ile Ala Ile Lys Asn Ser Arg Ala Ser Ser Val Ser Arg Val Val 
                85                  90                  95      


Asp Pro Thr Val Ile Val Lys Gly Asn Lys Leu Tyr Ile Leu Val Gly 
            100                 105                 110         


Ser Phe Asn Lys Thr Arg Asn Tyr Trp Thr Gln His Arg Asp Gly Ser 
        115                 120                 125             


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


Asn Gly Lys Thr Thr Ala Thr Ile Ser Trp Gly Lys Pro Val Ser Leu 
145                 150                 155                 160 


Lys Pro Leu Phe Pro Ala Glu Phe Asp Gly Ile Leu Thr Lys Glu Phe 
                165                 170                 175     


Val Gly Gly Val Gly Ala Ala Ile Val Ala Ser Asn Gly Asn Leu Val 
            180                 185                 190         


Tyr Pro Val Gln Val Thr Asn Lys Lys Lys Gln Val Phe Thr Lys Ile 
        195                 200                 205             


Met Tyr Ser Glu Asp Asp Gly Asn Thr Trp Lys Phe Ala Glu Gly Arg 
    210                 215                 220                 


Ser Lys Phe Gly Cys Ser Glu Pro Ala Val Leu Glu Trp Glu Gly Lys 
225                 230                 235                 240 


Leu Ile Ile Asn Asn Arg Val Asp Tyr Asn Arg Arg Leu Val Tyr Glu 
                245                 250                 255     


Ser Ser Asp Met Gly Lys Thr Trp Val Glu Ala Leu Gly Thr Leu Ser 
            260                 265                 270         


His Val Trp Thr Asn Ser Pro Thr Ser Asn Gln Pro Asp Cys Gln Ser 
        275                 280                 285             


Ser Phe Val Ala Val Thr Ile Glu Gly Lys Arg Val Met Leu Phe Thr 
    290                 295                 300                 


His Pro Leu Asn Leu Lys Gly Arg Trp Met Arg Asp Arg Leu His Leu 
305                 310                 315                 320 


Trp Met Thr Asp Asn Gln Arg Ile Phe Asp Val Gly Gln Ile Ser Ile 
                325                 330                 335     


Gly Asp Glu Asn Ser Gly Tyr Ser Ser Val Leu Tyr Lys Asp Asp Lys 
            340                 345                 350         


Leu Tyr Ser Leu His Glu Ile Asn Thr Asn Asp Val Tyr Ser Leu Val 
        355                 360                 365             


Phe Val Arg Leu Ile Gly Glu Leu Gln Leu Met Lys Ser Val Val Arg 
    370                 375                 380                 


Thr Trp Lys Glu Glu Asp Asn His Leu Ala Ser Ile Cys Thr Pro Val 
385                 390                 395                 400 


Val Pro Ala Thr Pro Pro Ser Lys Gly Ala Cys Gly Ala Ala Val Pro 
                405                 410                 415     


Thr Ala Gly Leu Val Gly Phe Leu Ser His Ser Ala Asn Gly Ser Val 
            420                 425                 430         


Trp Glu Asp Val Tyr Arg Cys Val Asp Ala Asn Val Ala Asn Ala Glu 
        435                 440                 445             


Arg Val Pro Asn Gly Leu Lys Phe Asn Gly Val Gly Gly Gly Ala Val 
    450                 455                 460                 


Trp Pro Val Ala Arg Gln Gly Gln Thr Arg Arg Tyr Gln Phe Ala Asn 
465                 470                 475                 480 


Tyr Arg Phe Thr Leu Val Ala Thr Val Thr Ile Asp Glu Leu Pro Lys 
                485                 490                 495     


Gly Thr Ser Pro Leu Leu Gly Ala Gly Leu Glu Gly Pro Gly Asp Ala 
            500                 505                 510         


Lys Leu Leu Gly Leu Ser Tyr Asp Lys Asn Arg Gln Trp Arg Pro Leu 
        515                 520                 525             


Tyr Gly Ala Ala Pro Ala Ser Pro Thr Gly Ser Trp Glu Leu His Lys 
    530                 535                 540                 


Lys Tyr His Val Val Leu Thr Met Ala Asp Arg Gln Gly Ser Val Tyr 
545                 550                 555                 560 


Val Asp Gly Gln Pro Leu Ala Gly Ser Gly Asn Thr Val Val Arg Gly 
                565                 570                 575     


Ala Thr Leu Pro Asp Ile Ser His Phe Tyr Ile Gly Gly Pro Arg Ser 
            580                 585                 590         


Lys Gly Ala Pro Thr Asp Ser Arg Val Thr Val Thr Asn Val Val Leu 
        595                 600                 605             


Tyr Asn Arg Arg Leu Asn Ser Ser Glu Ile Arg Thr Leu Phe Leu Ser 
    610                 615                 620                 


Gln Asp Met Ile Gly Thr Asp Gly Gly Ala Gly Thr Ala Ala 
625                 630                 635             


<210>  5
<211>  345
<212>  PRT
<213>  Trypanosoma cruzi


<220>
<221>  PEPTIDE
<222>  (1)..(345)
<223>  Catalytic domain of T. cruzi trans-sialidase [Q26966]

<400>  5

Thr Glu Arg Val Val His Ser Phe Arg Leu Pro Ala Leu Val Asn Val 
1               5                   10                  15      


Asp Gly Val Met Val Ala Ile Ala Asp Ala Arg Tyr Glu Thr Ser Asn 
            20                  25                  30          


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


Glu Thr Trp Glu Thr Gln Ile Ala Ile Lys Asn Ser Arg Ala Ser Ser 
    50                  55                  60                  


Val Ser Arg Val Val Asp Pro Thr Val Ile Val Lys Gly Asn Lys Leu 
65                  70                  75                  80  


Tyr Val Leu Val Gly Ser Tyr Asn Ser Ser Arg Ser Tyr Trp Thr Ser 
                85                  90                  95      


His Gly Asp Ala Arg Asp Trp Asp Ile Leu Leu Ala Val Gly Glu Val 
            100                 105                 110         


Thr Lys Ser Thr Ala Gly Gly Lys Ile Thr Ala Ser Ile Lys Trp Gly 
        115                 120                 125             


Ser Pro Val Ser Leu Lys Glu Phe Phe Pro Ala Glu Met Glu Gly Met 
    130                 135                 140                 


His Thr Asn Gln Phe Leu Gly Gly Ala Gly Val Ala Ile Val Ala Ser 
145                 150                 155                 160 


Asn Gly Asn Leu Val Tyr Pro Val Gln Val Thr Asn Lys Lys Lys Gln 
                165                 170                 175     


Val Phe Ser Lys Ile Phe Tyr Ser Glu Asp Glu Gly Lys Thr Trp Lys 
            180                 185                 190         


Phe Gly Lys Gly Arg Ser Ala Phe Gly Cys Ser Glu Pro Val Ala Leu 
        195                 200                 205             


Glu Trp Glu Gly Lys Leu Ile Ile Asn Thr Arg Val Asp Tyr Arg Arg 
    210                 215                 220                 


Arg Leu Val Tyr Glu Ser Ser Asp Met Gly Asn Ser Trp Leu Glu Ala 
225                 230                 235                 240 


Val Gly Thr Leu Ser Arg Val Trp Gly Pro Ser Pro Lys Ser Asn Gln 
                245                 250                 255     


Pro Gly Ser Gln Ser Ser Phe Thr Ala Val Thr Ile Glu Gly Met Arg 
            260                 265                 270         


Val Met Leu Phe Thr His Pro Leu Asn Phe Lys Gly Arg Trp Leu Arg 
        275                 280                 285             


Asp Arg Leu Asn Leu Trp Leu Thr Asp Asn Gln Arg Ile Tyr Asn Val 
    290                 295                 300                 


Gly Gln Val Ser Ile Gly Asp Glu Asn Ser Ala Tyr Ser Ser Val Leu 
305                 310                 315                 320 


Tyr Lys Asp Asp Lys Leu Tyr Cys Leu His Glu Ile Asn Ser Asn Glu 
                325                 330                 335     


Val Tyr Ser Leu Val Phe Ala Arg Leu 
            340                 345 


<210>  6
<211>  347
<212>  PRT
<213>  Trypanosoma congolense


<220>
<221>  PEPTIDE
<222>  (1)..(347)
<223>  Catalytic domain of T. congolese trans-sialidase [G0WJG3]

<400>  6

Gly Thr Thr Met Arg Thr Val His Ser Tyr Arg Ile Pro Ser Ile Val 
1               5                   10                  15      


Glu Val Gly Gly Val Leu Met Cys Val Gly Asp Ala Arg Tyr Ile Thr 
            20                  25                  30          


Ser Thr Asp Tyr Phe Phe Thr Asp Thr Val Ala Ala Tyr Ser Thr Asp 
        35                  40                  45              


Gly Gly Arg Thr Trp Lys Arg Glu Val Ile Ile Pro Asn Gly Arg Val 
    50                  55                  60                  


Asp Ala His Tyr Ser Arg Val Val Asp Pro Thr Val Val Ala Lys Gly 
65                  70                  75                  80  


Asn Asn Ile Tyr Val Leu Val Gly Arg Tyr Asn Val Thr Arg Gly Tyr 
                85                  90                  95      


Trp His Asn Lys Asn Asn Arg Ala Gly Val Ala Asp Trp Glu Pro Phe 
            100                 105                 110         


Val Tyr Lys Gly Thr Val Asn Val Gly Thr Lys Asp Asn Ala Thr Asp 
        115                 120                 125             


Val Ser Ile Ser Trp Glu Arg Thr Ala Leu Lys Ser Leu Tyr Asn Phe 
    130                 135                 140                 


Pro Val Ser Gly Ser Pro Gly Thr Gln Phe Leu Gly Gly Ala Gly Gly 
145                 150                 155                 160 


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


Asn Lys Ala Asn Arg Val Val Ser Met Ile Leu Tyr Ser Ala Asp Asp 
            180                 185                 190         


Gly Lys Ser Trp His Phe Gly Lys Gly Glu Ala Gly Val Gly Thr Ser 
        195                 200                 205             


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


Ser Asp Gly Gly Gln Gly Tyr Arg Met Ile Phe Glu Ser Ser Asp Leu 
225                 230                 235                 240 


Gly Ala Thr Trp Lys Glu Met Leu Asn Ser Ile Ser Arg Val Ile Gly 
                245                 250                 255     


Asn Ser Pro Gly Arg Ser Gly Pro Gly Ser Ser Ser Gly Phe Ile Thr 
            260                 265                 270         


Val Thr Val Glu Gly Val Pro Val Met Leu Leu Thr His Pro Lys Asn 
        275                 280                 285             


Leu Lys Gly Ser Tyr Tyr Arg Asp Arg Leu Gln Met Trp Met Thr Asp 
    290                 295                 300                 


Gly Asn Arg Met Trp His Val Gly Gln Val Ser Glu Gly Asp Asp Asn 
305                 310                 315                 320 


Ser Ala Tyr Ser Ser Leu Leu Tyr Thr Pro Asp Gly Val Leu Tyr Cys 
                325                 330                 335     


Leu His Glu Gln Asn Ile Asp Glu Val Tyr Ser 
            340                 345         


<210>  7
<211>  355
<212>  PRT
<213>  Trypanosoma brucei


<220>
<221>  PEPTIDE
<222>  (1)..(355)
<223>  Catalytic domain of T. brucei trans-sialidase [Q57XJ2]

<400>  7

Arg Thr Val His Ser Phe Arg Ile Pro Ser Phe Val Glu Val Asp Gly 
1               5                   10                  15      


Val Leu Met Gly Ile Gly Asp Ala Arg Tyr Leu Thr Ser Thr Asp Tyr 
            20                  25                  30          


Phe Phe Thr Asp Thr Val Ala Lys Tyr Ser Ala Asp Gly Gly Lys Thr 
        35                  40                  45              


Trp Lys Thr Glu Val Ile Ile Glu Asn Gly Arg Val Asp Pro Thr Tyr 
    50                  55                  60                  


Ser Arg Val Val Asp Pro Thr Val Val Ala Lys Ala Asp Ser Val Phe 
65                  70                  75                  80  


Val Leu Val Ala Arg Tyr Asn Val Thr Lys Gly Tyr Trp His Asn Glu 
                85                  90                  95      


Asn Asn Ala Ala Gly Ile Ala Asp Trp Glu Pro Phe Met Tyr Lys Gly 
            100                 105                 110         


Val Val Thr Lys Gly Ala Asp Gly Lys Thr Ser Asp Val Arg Ile Ser 
        115                 120                 125             


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


Ser Lys Gly Thr Gln Phe Ile Gly Gly Ala Gly Asn Gly Val Val Thr 
145                 150                 155                 160 


Leu Asn Gly Thr Ile Leu Phe Pro Val Gln Ala Arg Asn Glu Asp Asn 
                165                 170                 175     


Ala Val Val Ser Met Val Met Tyr Ser Val Asp Asp Gly Val Ser Trp 
            180                 185                 190         


His Phe Ala Arg Gly Glu Thr Ala Leu Leu Thr Ser Glu Ala Ser Leu 
        195                 200                 205             


Thr Glu Trp Asn Gly Lys Leu Leu Met Ser Ala Arg Thr Asp Thr Ser 
    210                 215                 220                 


Gly Val Asn Val Glu Gly Gly Phe Arg Lys Val Phe Glu Ser Asn Asn 
225                 230                 235                 240 


Leu Gly Ala Thr Trp Glu Glu Ser Leu Gly Thr Ile Ser Arg Val Ile 
                245                 250                 255     


Gly Asn Ser Pro Asp Arg Thr Lys Pro Ser Pro Thr Ala Asn Tyr Pro 
            260                 265                 270         


Gly Ser Ser Gly Ala Leu Ile Thr Val Thr Leu Gly Asp Val Pro Val 
        275                 280                 285             


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


Arg Leu Gln Leu Trp Met Thr Asp Gly Asn Arg Met Trp Leu Val Gly 
305                 310                 315                 320 


Gln Ile Ser Glu Gly Asp Asp Asn Ser Ala Tyr Ser Ser Leu Leu Leu 
                325                 330                 335     


Ala Arg Asp Gly Leu Leu Tyr Cys Leu His Glu Gln Asn Ile Asp Glu 
            340                 345                 350         


Val Tyr Gly 
        355 


<210>  8
<211>  270
<212>  PRT
<213>  Trypanosoma cruzi


<220>
<221>  PEPTIDE
<222>  (1)..(270)
<223>  C-terminal region (linker and CBD peptide) of T. cruzi 
       trans-sialidase [Q26966]

<400>  8

Val Gly Glu Leu Arg Ile Ile Lys Ser Val Leu Gln Ser Trp Lys Asn 
1               5                   10                  15      


Trp Asp Ser His Leu Ser Ser Ile Cys Thr Pro Ala Asp Pro Ala Ala 
            20                  25                  30          


Ser Ser Ser Glu Arg Gly Cys Gly Pro Ala Val Thr Thr Val Gly Leu 
        35                  40                  45              


Val Gly Phe Leu Ser His Ser Ala Thr Lys Thr Glu Trp Glu Asp Ala 
    50                  55                  60                  


Tyr Arg Cys Val Asn Ala Ser Thr Ala Asn Ala Glu Arg Val Pro Asn 
65                  70                  75                  80  


Gly Leu Lys Phe Ala Gly Val Gly Gly Gly Ala Leu Trp Pro Val Ser 
                85                  90                  95      


Gln Gln Gly Gln Asn Gln Arg Tyr Arg Phe Ala Asn His Ala Phe Thr 
            100                 105                 110         


Val Val Ala Ser Val Thr Ile His Glu Val Pro Ser Val Ala Ser Pro 
        115                 120                 125             


Leu Leu Gly Ala Ser Leu Asp Ser Ser Gly Gly Lys Lys Leu Leu Gly 
    130                 135                 140                 


Leu Ser Tyr Asp Glu Arg His Gln Trp Gln Pro Ile Tyr Gly Ser Thr 
145                 150                 155                 160 


Pro Val Thr Pro Thr Gly Ser Trp Glu Met Gly Lys Arg Tyr His Val 
                165                 170                 175     


Val Leu Thr Met Ala Asn Lys Ile Gly Ser Glu Tyr Ile Asp Gly Glu 
            180                 185                 190         


Pro Leu Glu Gly Ser Gly Gln Thr Val Val Pro Asp Glu Arg Thr Pro 
        195                 200                 205             


Asp Ile Ser His Phe Tyr Val Gly Gly Tyr Lys Arg Ser Asp Met Pro 
    210                 215                 220                 


Thr Ile Ser His Val Thr Val Asn Asn Val Leu Leu Tyr Asn Arg Gln 
225                 230                 235                 240 


Leu Asn Ala Glu Glu Ile Arg Thr Leu Phe Leu Ser Gln Asp Leu Ile 
                245                 250                 255     


Gly Thr Glu Ala His Met Asp Ser Ser Ser Asp Thr Ser Ala 
            260                 265                 270 


<210>  9
<211>  250
<212>  PRT
<213>  Trypanosoma congolense


<220>
<221>  PEPTIDE
<222>  (1)..(250)
<223>  C-terminal region (linker and CBD peptide) of T. congolese 
       trans-sialidase [G0WJG3]

<400>  9

Val Asp Glu Leu Lys Ser Ile Lys Ser Thr Ala Leu Val Trp Lys Ala 
1               5                   10                  15      


Gln Asp Glu Leu Leu Leu Gly Asn Cys Leu Pro Gly Asp Lys Tyr Asp 
            20                  25                  30          


Pro Gly Cys Asp Gly Ile Pro Thr Ala Gly Leu Ala Gly Leu Leu Val 
        35                  40                  45              


Gly Pro Leu Thr Glu Lys Thr Trp Pro Asp Ala Tyr Arg Cys Val Asn 
    50                  55                  60                  


Ala Ala Thr Ser Gly Ala Val Ser Thr Ala Glu Gly Val Arg Leu Asp 
65                  70                  75                  80  


Val Gly Gly Gly Gly His Val Val Trp Pro Val Ser Glu Gln Gly Gln 
                85                  90                  95      


Asp Gln Arg Tyr Tyr Phe Thr Asn Ser Glu Phe Thr Leu Ala Val Thr 
            100                 105                 110         


Val Arg Phe Asp Glu Met Pro Gln Gly Glu Leu Pro Leu Leu Gly Phe 
        115                 120                 125             


Val Asn Arg Glu Gly Lys Val Lys Lys Ile Leu Lys Val Ser Leu Ser 
    130                 135                 140                 


Gly Val Glu Trp Leu Leu Ala Tyr Gly Asn Glu Tyr Asn Ser Thr Ala 
145                 150                 155                 160 


Ala Glu Pro Leu Asp Val Asn Glu Ser His Gln Val Val Leu Ala Leu 
                165                 170                 175     


His Asp Gly Ile Val Ser Leu His Val Asp Gly Gly Asn Thr Thr Ala 
            180                 185                 190         


Thr Val Ser Val Arg Val Ala Ser Pro Ala Glu Leu Leu Asn Ile His 
        195                 200                 205             


His Leu Phe Val Gly Thr Pro Val Asp Gly Gly Ala Lys Glu His Ala 
    210                 215                 220                 


Asn Ile Thr Val Ser Asn Val Leu Val Tyr Asn Arg Pro Leu Arg Gly 
225                 230                 235                 240 


Val Glu Leu Leu Gly Leu Phe Ala Asn Arg 
                245                 250 


<210>  10
<211>  284
<212>  PRT
<213>  Trypanosoma brucei


<220>
<221>  PEPTIDE
<222>  (1)..(284)
<223>  C-terminal region (linker and CBD peptide) of T. brucei 
       trans-sialidase [Q57XJ2]

<400>  10

Val Asp Glu Leu Glu Lys Val Asn Ala Thr Val Arg Lys Trp Lys Ala 
1               5                   10                  15      


Gln Asp Ala Leu Leu Ala Gly Leu Cys Ser Ser Ser Arg Lys Lys Asn 
            20                  25                  30          


Asp Pro Thr Cys Ser Gly Val Pro Thr Asp Gly Leu Val Gly Leu Leu 
        35                  40                  45              


Ala Gly Pro Val Gly Ala Ser Val Trp Ala Asp Val Tyr Asp Cys Val 
    50                  55                  60                  


Asn Ala Ser Ile Ser Asp Gly Val Lys Val Ser Glu Gly Val Gln Leu 
65                  70                  75                  80  


Gly Gly Lys Arg Asn Ser Arg Val Leu Trp Pro Val Ser Glu Gln Gly 
                85                  90                  95      


Gln Asp Gln Arg Tyr Tyr Phe Ala Asn Thr His Phe Thr Leu Leu Ala 
            100                 105                 110         


Thr Val Arg Phe Ala Gly Glu Pro Lys Ala Glu Ala Pro Leu Met Gly 
        115                 120                 125             


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


Gly Gly Lys Lys Trp Val Leu Thr Tyr Gly Ser Val Arg Lys Glu Gly 
145                 150                 155                 160 


Pro Thr Thr Ser Met Asp Trp Asn Gln Thr His Gln Ile Ala Leu Thr 
                165                 170                 175     


Leu Arg Asp Gly Lys Val Asp Ala His Val Asn Gly Glu Leu Ile Ile 
            180                 185                 190         


Lys Glu Val Ser Val Gly Ala Ser Glu Ser Ser Ala His Leu His Leu 
        195                 200                 205             


Ser His Phe Phe Ile Gly Ala Pro Val Asn Asp Ser Gly Glu Gly Gly 
    210                 215                 220                 


Asn Asn Val Ile Val Arg Asn Val Leu Leu Tyr Asn Arg Lys Leu Asp 
225                 230                 235                 240 


Glu Asp Glu Leu Gln Leu Leu Tyr Ser Asn Arg Glu Lys Ile Gln Pro 
                245                 250                 255     


Val Val Ser Ala Val Gly Ile Pro Glu Gly Met Ser Ala Pro Arg Leu 
            260                 265                 270         


Cys Cys Leu Leu Ile Leu Met Tyr Val Leu Ala Ile 
        275                 280                 


<210>  11
<211>  2259
<212>  DNA
<213>  Trypanosoma rangeli


<220>
<221>  CDS
<222>  (1)..(273)
<223>  Nt 1-273 encode a-factor signal sequence; Kex2 and Ste3 sites;

<220>
<221>  CDS
<222>  (274)..(2187)
<223>  Nt 274-2187 encode Tr13 mutant trans-sialidase;

<220>
<221>  CDS
<222>  (2188)..(2259)
<223>  Nt 274-2187 encode a C-terminal c-myc-tag and His-tag.;

<400>  11
atg aga ttt cct tca att ttt act gct gtt tta ttc gca gca tcc tcc         48
Met Arg Phe Pro Ser Ile Phe Thr Ala Val Leu Phe Ala Ala Ser Ser           
1               5                   10                  15                

gca tta gct gct cca gtc aac act aca aca gaa gat gaa acg gca caa         96
Ala Leu Ala Ala Pro Val Asn Thr Thr Thr Glu Asp Glu Thr Ala Gln           
            20                  25                  30                    

att ccg gct gaa gct gtc atc ggt tac tca gat tta gaa ggg gat ttc        144
Ile Pro Ala Glu Ala Val Ile Gly Tyr Ser Asp Leu Glu Gly Asp Phe           
        35                  40                  45                        

gat gtt gct gtt ttg cca ttt tcc aac agc aca aat aac ggg tta ttg        192
Asp Val Ala Val Leu Pro Phe Ser Asn Ser Thr Asn Asn Gly Leu Leu           
    50                  55                  60                            

ttt ata aat act act att gcc agc att gct gct aaa gaa gaa ggg gta        240
Phe Ile Asn Thr Thr Ile Ala Ser Ile Ala Ala Lys Glu Glu Gly Val           
65                  70                  75                  80            

tct ctc gag aag aga gag gct gaa gct gct tct ttg gct ccc gga tca        288
Ser Leu Glu Lys Arg Glu Ala Glu Ala Ala Ser Leu Ala Pro Gly Ser           
                85                  90                  95                

tct cgt gtg gaa tta ttt aaa aga aaa aac tcc acc gtg cca ttt gag        336
Ser Arg Val Glu Leu Phe Lys Arg Lys Asn Ser Thr Val Pro Phe Glu           
            100                 105                 110                   

gag tca aac ggt act ata cga gaa cgt gtg gtt cat tca ttt aga tta        384
Glu Ser Asn Gly Thr Ile Arg Glu Arg Val Val His Ser Phe Arg Leu           
        115                 120                 125                       

cca act atc gtt aac gta gat gga gtc atg gtt gcc att gct gat gcc        432
Pro Thr Ile Val Asn Val Asp Gly Val Met Val Ala Ile Ala Asp Ala           
    130                 135                 140                           

aga tat gag aca tca ttc gac aac tcc ttt atc gaa act gct gtt aaa        480
Arg Tyr Glu Thr Ser Phe Asp Asn Ser Phe Ile Glu Thr Ala Val Lys           
145                 150                 155                 160           

tac agt gtt gac gat ggt gct acg tgg aat aca caa att gca atc aaa        528
Tyr Ser Val Asp Asp Gly Ala Thr Trp Asn Thr Gln Ile Ala Ile Lys           
                165                 170                 175               

aat tct cgt gca tca tca gtt tca agg gtt gtc gat cct acg gtc ata        576
Asn Ser Arg Ala Ser Ser Val Ser Arg Val Val Asp Pro Thr Val Ile           
            180                 185                 190                   

gta aag gga aat aag ttg tat atc ctg gtt gga tcc ttt aac aag aca        624
Val Lys Gly Asn Lys Leu Tyr Ile Leu Val Gly Ser Phe Asn Lys Thr           
        195                 200                 205                       

agg aac tat tgg acc cag cac aga gat gga tct gac tgg gaa cca ttg        672
Arg Asn Tyr Trp Thr Gln His Arg Asp Gly Ser Asp Trp Glu Pro Leu           
    210                 215                 220                           

ttg gtg gtt gga gag gtt acg aag tct gct gct aac ggt aaa aca act        720
Leu Val Val Gly Glu Val Thr Lys Ser Ala Ala Asn Gly Lys Thr Thr           
225                 230                 235                 240           

gca act att tca tgg ggg aaa cct gtc tcc ctt aag cct ttg ttc cct        768
Ala Thr Ile Ser Trp Gly Lys Pro Val Ser Leu Lys Pro Leu Phe Pro           
                245                 250                 255               

gca gag ttc gac ggc ata ctt act aag gaa ttc gta ggt gga gta ggc        816
Ala Glu Phe Asp Gly Ile Leu Thr Lys Glu Phe Val Gly Gly Val Gly           
            260                 265                 270                   

gcc gcc atc gtg gca agt aat ggt aat ttg gta tac cct gtg caa gta        864
Ala Ala Ile Val Ala Ser Asn Gly Asn Leu Val Tyr Pro Val Gln Val           
        275                 280                 285                       

act aat aag aag aag caa gta ttt aca aaa att atg tat tcc gag gat        912
Thr Asn Lys Lys Lys Gln Val Phe Thr Lys Ile Met Tyr Ser Glu Asp           
    290                 295                 300                           

gat ggt aac act tgg aag ttc gcc gaa gga agg tct aag ttc ggt tgc        960
Asp Gly Asn Thr Trp Lys Phe Ala Glu Gly Arg Ser Lys Phe Gly Cys           
305                 310                 315                 320           

tca gaa cca gca gtt ttg gaa tgg gaa gga aag cta atc att aat aac       1008
Ser Glu Pro Ala Val Leu Glu Trp Glu Gly Lys Leu Ile Ile Asn Asn           
                325                 330                 335               

cga gtc gat tac aat aga cgt ctg gtg tac gaa tcc agt gac atg ggc       1056
Arg Val Asp Tyr Asn Arg Arg Leu Val Tyr Glu Ser Ser Asp Met Gly           
            340                 345                 350                   

aaa aca tgg gta gag gct ctt ggt act ctg tcc cac gtc tgg acg aac       1104
Lys Thr Trp Val Glu Ala Leu Gly Thr Leu Ser His Val Trp Thr Asn           
        355                 360                 365                       

agt cca act tcc aat caa ccc gat tgt cag agt tca ttc gtt gca gtt       1152
Ser Pro Thr Ser Asn Gln Pro Asp Cys Gln Ser Ser Phe Val Ala Val           
    370                 375                 380                           

act atc gaa ggt aaa cga gtg atg ttg ttt act cat cca cta aat ttg       1200
Thr Ile Glu Gly Lys Arg Val Met Leu Phe Thr His Pro Leu Asn Leu           
385                 390                 395                 400           

aag ggt aga tgg atg agg gat aga ctt cat ctg tgg atg acc gat aat       1248
Lys Gly Arg Trp Met Arg Asp Arg Leu His Leu Trp Met Thr Asp Asn           
                405                 410                 415               

cag aga atc ttt gat gtt ggc caa att tcc att ggt gat gaa aac agt       1296
Gln Arg Ile Phe Asp Val Gly Gln Ile Ser Ile Gly Asp Glu Asn Ser           
            420                 425                 430                   

ggt tac tct tcc gtc cta tac aag gac gat aaa tta tat tcc cta cat       1344
Gly Tyr Ser Ser Val Leu Tyr Lys Asp Asp Lys Leu Tyr Ser Leu His           
        435                 440                 445                       

gag att aat act aat gat gtt tat tct ctt gtt ttt gtc cga ttg att       1392
Glu Ile Asn Thr Asn Asp Val Tyr Ser Leu Val Phe Val Arg Leu Ile           
    450                 455                 460                           

ggt gag ctg cag tta atg aaa agt gtg gtt cgt acc tgg aag gaa gag       1440
Gly Glu Leu Gln Leu Met Lys Ser Val Val Arg Thr Trp Lys Glu Glu           
465                 470                 475                 480           

gac aat cat ttg gct tca ata tgt act cca gtc gta cca gca acc cca       1488
Asp Asn His Leu Ala Ser Ile Cys Thr Pro Val Val Pro Ala Thr Pro           
                485                 490                 495               

cca agt aaa gga gcc tgc ggt gcc gct gta cct aca gct ggt tta gtt       1536
Pro Ser Lys Gly Ala Cys Gly Ala Ala Val Pro Thr Ala Gly Leu Val           
            500                 505                 510                   

ggc ttc tta tct cac tca gct aat gga tcc gtt tgg gag gac gta tat       1584
Gly Phe Leu Ser His Ser Ala Asn Gly Ser Val Trp Glu Asp Val Tyr           
        515                 520                 525                       

aga tgt gtc gat gct aac gtc gcc aac gct gag aga gtt cct aac ggc       1632
Arg Cys Val Asp Ala Asn Val Ala Asn Ala Glu Arg Val Pro Asn Gly           
    530                 535                 540                           

ctt aag ttt aat ggg gtt ggt ggg ggc gct gtc tgg cca gtc gcc agg       1680
Leu Lys Phe Asn Gly Val Gly Gly Gly Ala Val Trp Pro Val Ala Arg           
545                 550                 555                 560           

cag gga caa acc cga agg tac caa ttc gca aac tac aga ttt acc tta       1728
Gln Gly Gln Thr Arg Arg Tyr Gln Phe Ala Asn Tyr Arg Phe Thr Leu           
                565                 570                 575               

gtc gcc acc gtt acg att gac gaa ttg ccc aaa ggt acc tct ccc ctt       1776
Val Ala Thr Val Thr Ile Asp Glu Leu Pro Lys Gly Thr Ser Pro Leu           
            580                 585                 590                   

ctt ggt gcc ggg tta gaa ggt cca ggc gac gct aaa ttg cta ggt tta       1824
Leu Gly Ala Gly Leu Glu Gly Pro Gly Asp Ala Lys Leu Leu Gly Leu           
        595                 600                 605                       

tct tac gac aag aac cgt caa tgg cga ccc ttg tac gga gca gcc cct       1872
Ser Tyr Asp Lys Asn Arg Gln Trp Arg Pro Leu Tyr Gly Ala Ala Pro           
    610                 615                 620                           

gct tct cct aca gga tct tgg gag cta cac aag aag tac cat gta gtc       1920
Ala Ser Pro Thr Gly Ser Trp Glu Leu His Lys Lys Tyr His Val Val           
625                 630                 635                 640           

ctg acc atg gct gac aga cag ggg agt gtt tat gtt gat ggg caa cct       1968
Leu Thr Met Ala Asp Arg Gln Gly Ser Val Tyr Val Asp Gly Gln Pro           
                645                 650                 655               

ctt gcc gga tca ggc aat acc gtg gtt aga gga gct act ttg cca gac       2016
Leu Ala Gly Ser Gly Asn Thr Val Val Arg Gly Ala Thr Leu Pro Asp           
            660                 665                 670                   

atc tct cac ttc tac att ggt gga ccc aga tct aag gga gcc cct aca       2064
Ile Ser His Phe Tyr Ile Gly Gly Pro Arg Ser Lys Gly Ala Pro Thr           
        675                 680                 685                       

gat agt aga gtt aca gtg act aac gtg gtt tta tat aat cgt aga ctg       2112
Asp Ser Arg Val Thr Val Thr Asn Val Val Leu Tyr Asn Arg Arg Leu           
    690                 695                 700                           

aac tct agt gag atc cgt aca ctt ttt ttg tct caa gac atg ata ggt       2160
Asn Ser Ser Glu Ile Arg Thr Leu Phe Leu Ser Gln Asp Met Ile Gly           
705                 710                 715                 720           

acc gat ggt gga gct ggt aca gca gca ttt cta gaa caa aaa ctc atc       2208
Thr Asp Gly Gly Ala Gly Thr Ala Ala Phe Leu Glu Gln Lys Leu Ile           
                725                 730                 735               

tca gaa gag gat ctg aat agc gcc gtc gac cat cat cat cat cat cat       2256
Ser Glu Glu Asp Leu Asn Ser Ala Val Asp His His His His His His           
            740                 745                 750                   

tga                                                                   2259


<210>  12
<211>  91
<212>  PRT
<213>  Trypanosoma rangeli

<400>  12

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


Ala Leu Ala Ala Pro Val Asn Thr Thr Thr Glu Asp Glu Thr Ala Gln 
            20                  25                  30          


Ile Pro Ala Glu Ala Val Ile Gly Tyr Ser Asp Leu Glu Gly Asp Phe 
        35                  40                  45              


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


Phe Ile Asn Thr Thr Ile Ala Ser Ile Ala Ala Lys Glu Glu Gly Val 
65                  70                  75                  80  


Ser Leu Glu Lys Arg Glu Ala Glu Ala Ala Ser 
                85                  90      


<210>  13
<211>  638
<212>  PRT
<213>  Trypanosoma rangeli

<400>  13

Leu Ala Pro Gly Ser Ser Arg Val Glu Leu Phe Lys Arg Lys Asn Ser 
1               5                   10                  15      


Thr Val Pro Phe Glu Glu Ser Asn Gly Thr Ile Arg Glu Arg Val Val 
            20                  25                  30          


His Ser Phe Arg Leu Pro Thr Ile Val Asn Val Asp Gly Val Met Val 
        35                  40                  45              


Ala Ile Ala Asp Ala Arg Tyr Glu Thr Ser Phe Asp Asn Ser Phe Ile 
    50                  55                  60                  


Glu Thr Ala Val Lys Tyr Ser Val Asp Asp Gly Ala Thr Trp Asn Thr 
65                  70                  75                  80  


Gln Ile Ala Ile Lys Asn Ser Arg Ala Ser Ser Val Ser Arg Val Val 
                85                  90                  95      


Asp Pro Thr Val Ile Val Lys Gly Asn Lys Leu Tyr Ile Leu Val Gly 
            100                 105                 110         


Ser Phe Asn Lys Thr Arg Asn Tyr Trp Thr Gln His Arg Asp Gly Ser 
        115                 120                 125             


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


Asn Gly Lys Thr Thr Ala Thr Ile Ser Trp Gly Lys Pro Val Ser Leu 
145                 150                 155                 160 


Lys Pro Leu Phe Pro Ala Glu Phe Asp Gly Ile Leu Thr Lys Glu Phe 
                165                 170                 175     


Val Gly Gly Val Gly Ala Ala Ile Val Ala Ser Asn Gly Asn Leu Val 
            180                 185                 190         


Tyr Pro Val Gln Val Thr Asn Lys Lys Lys Gln Val Phe Thr Lys Ile 
        195                 200                 205             


Met Tyr Ser Glu Asp Asp Gly Asn Thr Trp Lys Phe Ala Glu Gly Arg 
    210                 215                 220                 


Ser Lys Phe Gly Cys Ser Glu Pro Ala Val Leu Glu Trp Glu Gly Lys 
225                 230                 235                 240 


Leu Ile Ile Asn Asn Arg Val Asp Tyr Asn Arg Arg Leu Val Tyr Glu 
                245                 250                 255     


Ser Ser Asp Met Gly Lys Thr Trp Val Glu Ala Leu Gly Thr Leu Ser 
            260                 265                 270         


His Val Trp Thr Asn Ser Pro Thr Ser Asn Gln Pro Asp Cys Gln Ser 
        275                 280                 285             


Ser Phe Val Ala Val Thr Ile Glu Gly Lys Arg Val Met Leu Phe Thr 
    290                 295                 300                 


His Pro Leu Asn Leu Lys Gly Arg Trp Met Arg Asp Arg Leu His Leu 
305                 310                 315                 320 


Trp Met Thr Asp Asn Gln Arg Ile Phe Asp Val Gly Gln Ile Ser Ile 
                325                 330                 335     


Gly Asp Glu Asn Ser Gly Tyr Ser Ser Val Leu Tyr Lys Asp Asp Lys 
            340                 345                 350         


Leu Tyr Ser Leu His Glu Ile Asn Thr Asn Asp Val Tyr Ser Leu Val 
        355                 360                 365             


Phe Val Arg Leu Ile Gly Glu Leu Gln Leu Met Lys Ser Val Val Arg 
    370                 375                 380                 


Thr Trp Lys Glu Glu Asp Asn His Leu Ala Ser Ile Cys Thr Pro Val 
385                 390                 395                 400 


Val Pro Ala Thr Pro Pro Ser Lys Gly Ala Cys Gly Ala Ala Val Pro 
                405                 410                 415     


Thr Ala Gly Leu Val Gly Phe Leu Ser His Ser Ala Asn Gly Ser Val 
            420                 425                 430         


Trp Glu Asp Val Tyr Arg Cys Val Asp Ala Asn Val Ala Asn Ala Glu 
        435                 440                 445             


Arg Val Pro Asn Gly Leu Lys Phe Asn Gly Val Gly Gly Gly Ala Val 
    450                 455                 460                 


Trp Pro Val Ala Arg Gln Gly Gln Thr Arg Arg Tyr Gln Phe Ala Asn 
465                 470                 475                 480 


Tyr Arg Phe Thr Leu Val Ala Thr Val Thr Ile Asp Glu Leu Pro Lys 
                485                 490                 495     


Gly Thr Ser Pro Leu Leu Gly Ala Gly Leu Glu Gly Pro Gly Asp Ala 
            500                 505                 510         


Lys Leu Leu Gly Leu Ser Tyr Asp Lys Asn Arg Gln Trp Arg Pro Leu 
        515                 520                 525             


Tyr Gly Ala Ala Pro Ala Ser Pro Thr Gly Ser Trp Glu Leu His Lys 
    530                 535                 540                 


Lys Tyr His Val Val Leu Thr Met Ala Asp Arg Gln Gly Ser Val Tyr 
545                 550                 555                 560 


Val Asp Gly Gln Pro Leu Ala Gly Ser Gly Asn Thr Val Val Arg Gly 
                565                 570                 575     


Ala Thr Leu Pro Asp Ile Ser His Phe Tyr Ile Gly Gly Pro Arg Ser 
            580                 585                 590         


Lys Gly Ala Pro Thr Asp Ser Arg Val Thr Val Thr Asn Val Val Leu 
        595                 600                 605             


Tyr Asn Arg Arg Leu Asn Ser Ser Glu Ile Arg Thr Leu Phe Leu Ser 
    610                 615                 620                 


Gln Asp Met Ile Gly Thr Asp Gly Gly Ala Gly Thr Ala Ala 
625                 630                 635             


<210>  14
<211>  23
<212>  PRT
<213>  Trypanosoma rangeli

<400>  14

Phe Leu Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu Asn Ser Ala Val 
1               5                   10                  15      


Asp His His His His His His 
            20              


<210>  15
<211>  25
<212>  DNA
<213>  Trypanosoma rangeli


<220>
<221>  primer_bind
<222>  (1)..(25)
<223>  Tr_fwd PCR primer - Tr6 trans-sialidase gene

<400>  15
gctctcgaga agagagaggc tgaag                                             25


<210>  16
<211>  26
<212>  DNA
<213>  Trypanosoma rangeli


<220>
<221>  primer_bind
<222>  (1)..(26)
<223>  Tr_rev PCR primer - Tr6 trans-sialidase gene

<400>  16
cgctctagaa atgctgctgt accagc                                            26


<210>  17
<211>  34
<212>  DNA
<213>  Trypanosoma rangeli


<220>
<221>  primer_bind
<222>  (1)..(34)
<223>  Q123S_F mutation primer for Tr6 gene

<400>  17
ctattggacc tctcacagag atggatctga ctgg                                   34


<210>  18
<211>  35
<212>  DNA
<213>  Trypanosoma rangeli


<220>
<221>  primer_bind
<222>  (1)..(35)
<223>  Q123S_R mutation primer for Tr6 gene

<400>  18
catctctgtg agaggtccaa tagttccttg tcttg                                  35


<210>  19
<211>  33
<212>  DNA
<213>  Trypanosoma rangeli


<220>
<221>  primer_bind
<222>  (1)..(33)
<223>  R125G_F mutation primer for Tr6 gene

<400>  19
gacccagcac ggagatggat ctgactggga acc                                    33


<210>  20
<211>  33
<212>  DNA
<213>  Trypanosoma rangeli


<220>
<221>  primer_bind
<222>  (1)..(33)
<223>  R125G_R mutation primer for Tr6 gene

<400>  20
cagatccatc tccgtgctgg gtccaatagt tcc                                    33


<210>  21
<211>  34
<212>  DNA
<213>  Trypanosoma rangeli


<220>
<221>  primer_bind
<222>  (1)..(34)
<223>  G127A_F mutation primer for Tr6 gene

<400>  21
gcacagagat gcttctgact gggaaccatt gttg                                   34


<210>  22
<211>  35
<212>  DNA
<213>  Trypanosoma rangeli


<220>
<221>  primer_bind
<222>  (1)..(35)
<223>  G127A_R mutation primer for Tr6 gene

<400>  22
cccagtcaga agcatctctg tgctgggtcc aatag                                  35


<210>  23
<211>  32
<212>  DNA
<213>  Trypanosoma rangeli


<220>
<221>  primer_bind
<222>  (1)..(32)
<223>  E175Q_F mutation primer for Tr6 gene

<400>  23
acttactaag cagttcgtag gtggagtagg cg                                     32


<210>  24
<211>  40
<212>  DNA
<213>  Trypanosoma rangeli


<220>
<221>  primer_bind
<222>  (1)..(40)
<223>  E175Q_R mutation primer for Tr6 gene

<400>  24
ctccacctac gaactgctta gtaagtatgc cgtcgaactc                             40


<210>  25
<211>  29
<212>  DNA
<213>  Trypanosoma rangeli


<220>
<221>  primer_bind
<222>  (1)..(29)
<223>  V177L_F mutation primer for Tr6 gene

<400>  25
taaggaattc ttgggtggag taggcgccg                                         29


<210>  26
<211>  39
<212>  DNA
<213>  Trypanosoma rangeli


<220>
<221>  primer_bind
<222>  (1)..(39)
<223>  V177L_R mutation primer for Tr6 gene

<400>  26
cctactccac ccaagaattc cttagtaagt atgccgtcg                              39


<210>  27
<211>  28
<212>  DNA
<213>  Trypanosoma rangeli


<220>
<221>  primer_bind
<222>  (1)..(28)
<223>  V180A_F mutation primer for Tr6 gene

<400>  27
cgtaggtgga gctggcgccg ccatcgtg                                          28


<210>  28
<211>  37
<212>  DNA
<213>  Trypanosoma rangeli


<220>
<221>  primer_bind
<222>  (1)..(37)
<223>  V180A_R mutation primer for Tr6 gene

<400>  28
tggcggcgcc agctccacct acgaattcct tagtaag                                37


<210>  29
<211>  43
<212>  DNA
<213>  Trypanosoma rangeli


<220>
<221>  primer_bind
<222>  (1)..(43)
<223>  G202K_F mutation primer for Tr6 gene

<400>  29
tgctgacatg aagggaagag tatttacaaa aattatgtat tcc                         43


<210>  30
<211>  33
<212>  DNA
<213>  Trypanosoma rangeli


<220>
<221>  primer_bind
<222>  (1)..(33)
<223>  G202K_R mutation primer for Tr6 gene

<400>  30
atactcttcc cttcatgtca gcaatttgca cag                                    33


<210>  31
<211>  34
<212>  DNA
<213>  Trypanosoma rangeli


<220>
<221>  primer_bind
<222>  (1)..(34)
<223>  N250R_F mutation primer for Tr6 gene

<400>  31
agtcgattac agaagacgtc tggtgtacga atcc                                   34


<210>  32
<211>  41
<212>  DNA
<213>  Trypanosoma rangeli


<220>
<221>  primer_bind
<222>  (1)..(41)
<223>  N250R_R mutation primer for Tr6 gene

<400>  32
ccagacgtct tctgtaatcg actcggttat taatgattag c                           41


<210>  33
<211>  41
<212>  DNA
<213>  Trypanosoma rangeli


<220>
<221>  primer_bind
<222>  (1)..(41)
<223>  D363E_F mutation primer for Tr6 gene

<400>  33
gagattaata ctaatgaggt ttattctctt gtttttgtcc g                           41


<210>  34
<211>  52
<212>  DNA
<213>  Trypanosoma rangeli


<220>
<221>  primer_bind
<222>  (1)..(52)
<223>  D363E_R mutation primer for Tr6 gene

<400>  34
caagagaata aacctcatta gtattaatct catgtaggga atataattta tc               52


<210>  35
<211>  59
<212>  DNA
<213>  Trypanosoma rangeli


<220>
<221>  primer_bind
<222>  (1)..(59)
<223>  VTNKKKQ 197-203 _F mutation primer for Tr6 gene

<400>  35
ccctgtgcaa gtaactaata agaagaagca agtatttaca aaaattatgt attccgagg        59


<210>  36
<211>  54
<212>  DNA
<213>  Trypanosoma rangeli


<220>
<221>  primer_bind
<222>  (1)..(54)
<223>  VTNKKKQ 197-203 _R mutation primer for Tr6 gene

<400>  36
ttgtaaatac ttgcttcttc ttattagtta cttgcacagg gtataccaaa ttac             54


<210>  37
<211>  40
<212>  DNA
<213>  Trypanosoma rangeli


<220>
<221>  primer_bind
<222>  (1)..(40)
<223>  P98A_F mutation primer for Tr6 gene

<400>  37
ggttgtcgat gctacggtca tagtaaaggg aaataagttg                             40


<210>  38
<211>  33
<212>  DNA
<213>  Trypanosoma rangeli


<220>
<221>  primer_bind
<222>  (1)..(33)
<223>  P98A_R mutation primer for Tr6 gene

<400>  38
ctatgaccgt agcatcgaca acccttgaaa ctg                                    33


<210>  39
<211>  36
<212>  DNA
<213>  Trypanosoma rangeli


<220>
<221>  primer_bind
<222>  (1)..(36)
<223>  Y249G_F mutation primer for Tr6 gene

<400>  39
ccgagtcgat ggaaatagac gtctggtgta cgaatc                                 36


<210>  40
<211>  38
<212>  DNA
<213>  Trypanosoma rangeli


<220>
<221>  primer_bind
<222>  (1)..(38)
<223>  Y249G_R mutation primer for Tr6 gene

<400>  40
gacgtctatt tccatcgact cggttattaa tgattagc                               38


