                               LISTADO DE SECUENCIAS

<110>  INSTITUTO DE SALUD CARLOS III; 
       UNIVERSIDAD CENTRAL DE VENEZUELA CARACAS
 
<120>  NUEVO INHIBIDOR DE SERINA PROTEASA Y SU USO

<130>  PCT-603

<150>  ES201031031
<151>  2010-07-03

<160>  9     

<170>  PatentIn version 3.3

<210>  1
<211>  1194
<212>  DNA
<213>  Anisakis simplex

<220>
<221>  CDS
<222>  (1)..(1191)
<223>  Secuencia codificante de AniSerp, inhibidor de serina proteasa

<220>
<221>  sig_peptide
<222>  (1)..(25)
<223>  Pptido seal

<400>  1
atg atg aca gca tta ccg ttt tta acg ctt ctg ggt atg ttc ttg ctg       48
Met Met Thr Ala Leu Pro Phe Leu Thr Leu Leu Gly Met Phe Leu Leu         
1               5                   10                  15              

atg gaa ggt gtc atc gtc gta caa ggt cag cag aca atc gat gat gcc       96
Met Glu Gly Val Ile Val Val Gln Gly Gln Gln Thr Ile Asp Asp Ala         
            20                  25                  30                  

caa gct gat ttt gca tgg aat tta ctt gaa gca agt gga ccc caa acc      144
Gln Ala Asp Phe Ala Trp Asn Leu Leu Glu Ala Ser Gly Pro Gln Thr         
        35                  40                  45                      

caa ttc ata gtg ctc tct cca ttc tca ata tca atc gca ctg gcg atg      192
Gln Phe Ile Val Leu Ser Pro Phe Ser Ile Ser Ile Ala Leu Ala Met         
    50                  55                  60                          

gtc tac gct ggt gct gaa gga aac aca aaa aca caa atc ggc aat acg      240
Val Tyr Ala Gly Ala Glu Gly Asn Thr Lys Thr Gln Ile Gly Asn Thr         
65                  70                  75                  80          

att gcg aaa ggt gtt aac aat gat agt gcg ctg att gat tat ttt tcg      288
Ile Ala Lys Gly Val Asn Asn Asp Ser Ala Leu Ile Asp Tyr Phe Ser         
                85                  90                  95              

tca gct gcc gaa caa atg gat aaa acc act gcg ggt ttc caa ctg aat      336
Ser Ala Ala Glu Gln Met Asp Lys Thr Thr Ala Gly Phe Gln Leu Asn         
            100                 105                 110                 

acg gcg aat aaa gtt tac att caa gat aaa ttc gag att ctc aaa agc      384
Thr Ala Asn Lys Val Tyr Ile Gln Asp Lys Phe Glu Ile Leu Lys Ser         
        115                 120                 125                     

ttc gag aat atc att aag caa aaa ttc gca agt gaa ata caa cag gtg      432
Phe Glu Asn Ile Ile Lys Gln Lys Phe Ala Ser Glu Ile Gln Gln Val         
    130                 135                 140                         

gat ttc gct gca aac aga gaa aca gca gag tta atc aat gca tgg gtt      480
Asp Phe Ala Ala Asn Arg Glu Thr Ala Glu Leu Ile Asn Ala Trp Val         
145                 150                 155                 160         

gag aag aaa acg aat tca cgc atc aga aat tta gtg cag cca caa atg      528
Glu Lys Lys Thr Asn Ser Arg Ile Arg Asn Leu Val Gln Pro Gln Met         
                165                 170                 175             

tta agc gct caa acc gct atg gca ctt atc aat gcg ata tat ttc aaa      576
Leu Ser Ala Gln Thr Ala Met Ala Leu Ile Asn Ala Ile Tyr Phe Lys         
            180                 185                 190                 

ggt gat tgg aaa act aaa ttc aat gcg gaa tca acc acc aag aaa atg      624
Gly Asp Trp Lys Thr Lys Phe Asn Ala Glu Ser Thr Thr Lys Lys Met         
        195                 200                 205                     

ttc cat gct ttc gaa aac tca gat cga gag gtt gat atg atg gca att      672
Phe His Ala Phe Glu Asn Ser Asp Arg Glu Val Asp Met Met Ala Ile         
    210                 215                 220                         

acg agt ggt ttc tcg tat gct gag act gag gat gtg caa gtg ctt gag      720
Thr Ser Gly Phe Ser Tyr Ala Glu Thr Glu Asp Val Gln Val Leu Glu         
225                 230                 235                 240         

cta ccg tat aaa gat cct gaa acg ttc atg tac gtt ttc cta cct act      768
Leu Pro Tyr Lys Asp Pro Glu Thr Phe Met Tyr Val Phe Leu Pro Thr         
                245                 250                 255             

gaa cgt ttc gga ctg agg caa ttc gag aag tcg atg aac ggc gag aaa      816
Glu Arg Phe Gly Leu Arg Gln Phe Glu Lys Ser Met Asn Gly Glu Lys         
            260                 265                 270                 

atc atg cag ttg atg aac gga tgc atg cca agg aat aaa att atc gtt      864
Ile Met Gln Leu Met Asn Gly Cys Met Pro Arg Asn Lys Ile Ile Val         
        275                 280                 285                     

gaa tta cca aaa ttc aag ttg gac ggt caa ttg aga cta aat aaa gtt      912
Glu Leu Pro Lys Phe Lys Leu Asp Gly Gln Leu Arg Leu Asn Lys Val         
    290                 295                 300                         

ctg gaa gaa ctg ggc att cgt gat gca ttc gat gcc gaa aaa gcg aac      960
Leu Glu Glu Leu Gly Ile Arg Asp Ala Phe Asp Ala Glu Lys Ala Asn         
305                 310                 315                 320         

ttc gat ggt atc agc aag aac gat tcg ttg gta gtg agt gat gtc gta     1008
Phe Asp Gly Ile Ser Lys Asn Asp Ser Leu Val Val Ser Asp Val Val         
                325                 330                 335             

cac aaa gcc aat gtt gag gtg aac gaa gaa ggc agt gaa gca gca gct     1056
His Lys Ala Asn Val Glu Val Asn Glu Glu Gly Ser Glu Ala Ala Ala         
            340                 345                 350                 

gct aca ggt cta ttt atg gta ttc cgc tcc agt cgt ccg atg ccg gtg     1104
Ala Thr Gly Leu Phe Met Val Phe Arg Ser Ser Arg Pro Met Pro Val         
        355                 360                 365                     

aca cca ccg ata aga ttc ata gct gat cat ccg ttc ata ttt acg atc     1152
Thr Pro Pro Ile Arg Phe Ile Ala Asp His Pro Phe Ile Phe Thr Ile         
    370                 375                 380                         

gtc agt cag aat cgc att ctg ttt att ggt cgt ttc cac tga             1194
Val Ser Gln Asn Arg Ile Leu Phe Ile Gly Arg Phe His                     
385                 390                 395                             


<210>  2
<211>  397
<212>  PRT
<213>  Anisakis simplex

<400>  2
Met Met Thr Ala Leu Pro Phe Leu Thr Leu Leu Gly Met Phe Leu Leu 
1               5                   10                  15      

Met Glu Gly Val Ile Val Val Gln Gly Gln Gln Thr Ile Asp Asp Ala 
            20                  25                  30          

Gln Ala Asp Phe Ala Trp Asn Leu Leu Glu Ala Ser Gly Pro Gln Thr 
        35                  40                  45              

Gln Phe Ile Val Leu Ser Pro Phe Ser Ile Ser Ile Ala Leu Ala Met 
    50                  55                  60                  

Val Tyr Ala Gly Ala Glu Gly Asn Thr Lys Thr Gln Ile Gly Asn Thr 
65                  70                  75                  80  

Ile Ala Lys Gly Val Asn Asn Asp Ser Ala Leu Ile Asp Tyr Phe Ser 
                85                  90                  95      

Ser Ala Ala Glu Gln Met Asp Lys Thr Thr Ala Gly Phe Gln Leu Asn 
            100                 105                 110         

Thr Ala Asn Lys Val Tyr Ile Gln Asp Lys Phe Glu Ile Leu Lys Ser 
        115                 120                 125             

Phe Glu Asn Ile Ile Lys Gln Lys Phe Ala Ser Glu Ile Gln Gln Val 
    130                 135                 140                 

Asp Phe Ala Ala Asn Arg Glu Thr Ala Glu Leu Ile Asn Ala Trp Val 
145                 150                 155                 160 

Glu Lys Lys Thr Asn Ser Arg Ile Arg Asn Leu Val Gln Pro Gln Met 
                165                 170                 175     

Leu Ser Ala Gln Thr Ala Met Ala Leu Ile Asn Ala Ile Tyr Phe Lys 
            180                 185                 190         

Gly Asp Trp Lys Thr Lys Phe Asn Ala Glu Ser Thr Thr Lys Lys Met 
        195                 200                 205             

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

Thr Ser Gly Phe Ser Tyr Ala Glu Thr Glu Asp Val Gln Val Leu Glu 
225                 230                 235                 240 

Leu Pro Tyr Lys Asp Pro Glu Thr Phe Met Tyr Val Phe Leu Pro Thr 
                245                 250                 255     

Glu Arg Phe Gly Leu Arg Gln Phe Glu Lys Ser Met Asn Gly Glu Lys 
            260                 265                 270         

Ile Met Gln Leu Met Asn Gly Cys Met Pro Arg Asn Lys Ile Ile Val 
        275                 280                 285             

Glu Leu Pro Lys Phe Lys Leu Asp Gly Gln Leu Arg Leu Asn Lys Val 
    290                 295                 300                 

Leu Glu Glu Leu Gly Ile Arg Asp Ala Phe Asp Ala Glu Lys Ala Asn 
305                 310                 315                 320 

Phe Asp Gly Ile Ser Lys Asn Asp Ser Leu Val Val Ser Asp Val Val 
                325                 330                 335     

His Lys Ala Asn Val Glu Val Asn Glu Glu Gly Ser Glu Ala Ala Ala 
            340                 345                 350         

Ala Thr Gly Leu Phe Met Val Phe Arg Ser Ser Arg Pro Met Pro Val 
        355                 360                 365             

Thr Pro Pro Ile Arg Phe Ile Ala Asp His Pro Phe Ile Phe Thr Ile 
    370                 375                 380                 

Val Ser Gln Asn Arg Ile Leu Phe Ile Gly Arg Phe His 
385                 390                 395         


<210>  3
<211>  372
<212>  PRT
<213>  Anisakis simplex


<220>
<221>  mat_peptide
<222>  (1)..(372)
<223>  Forma madura de AniSerp

<400>  3

Gln Gln Thr Ile Asp Asp Ala Gln Ala Asp Phe Ala Trp Asn Leu Leu 
1               5                   10                  15      

Glu Ala Ser Gly Pro Gln Thr Gln Phe Ile Val Leu Ser Pro Phe Ser 
            20                  25                  30          

Ile Ser Ile Ala Leu Ala Met Val Tyr Ala Gly Ala Glu Gly Asn Thr 
        35                  40                  45              

Lys Thr Gln Ile Gly Asn Thr Ile Ala Lys Gly Val Asn Asn Asp Ser 
    50                  55                  60                  

Ala Leu Ile Asp Tyr Phe Ser Ser Ala Ala Glu Gln Met Asp Lys Thr 
65                  70                  75                  80  

Thr Ala Gly Phe Gln Leu Asn Thr Ala Asn Lys Val Tyr Ile Gln Asp 
                85                  90                  95      

Lys Phe Glu Ile Leu Lys Ser Phe Glu Asn Ile Ile Lys Gln Lys Phe 
            100                 105                 110         

Ala Ser Glu Ile Gln Gln Val Asp Phe Ala Ala Asn Arg Glu Thr Ala 
        115                 120                 125             

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

Asn Leu Val Gln Pro Gln Met Leu Ser Ala Gln Thr Ala Met Ala Leu 
145                 150                 155                 160 

Ile Asn Ala Ile Tyr Phe Lys Gly Asp Trp Lys Thr Lys Phe Asn Ala 
                165                 170                 175     

Glu Ser Thr Thr Lys Lys Met Phe His Ala Phe Glu Asn Ser Asp Arg 
            180                 185                 190         

Glu Val Asp Met Met Ala Ile Thr Ser Gly Phe Ser Tyr Ala Glu Thr 
        195                 200                 205             

Glu Asp Val Gln Val Leu Glu Leu Pro Tyr Lys Asp Pro Glu Thr Phe 
    210                 215                 220                 

Met Tyr Val Phe Leu Pro Thr Glu Arg Phe Gly Leu Arg Gln Phe Glu 
225                 230                 235                 240 

Lys Ser Met Asn Gly Glu Lys Ile Met Gln Leu Met Asn Gly Cys Met 
                245                 250                 255     

Pro Arg Asn Lys Ile Ile Val Glu Leu Pro Lys Phe Lys Leu Asp Gly 
            260                 265                 270         

Gln Leu Arg Leu Asn Lys Val Leu Glu Glu Leu Gly Ile Arg Asp Ala 
        275                 280                 285             

Phe Asp Ala Glu Lys Ala Asn Phe Asp Gly Ile Ser Lys Asn Asp Ser 
    290                 295                 300                 

Leu Val Val Ser Asp Val Val His Lys Ala Asn Val Glu Val Asn Glu 
305                 310                 315                 320 

Glu Gly Ser Glu Ala Ala Ala Ala Thr Gly Leu Phe Met Val Phe Arg 
                325                 330                 335     

Ser Ser Arg Pro Met Pro Val Thr Pro Pro Ile Arg Phe Ile Ala Asp 
            340                 345                 350         

His Pro Phe Ile Phe Thr Ile Val Ser Gln Asn Arg Ile Leu Phe Ile 
        355                 360                 365             

Gly Arg Phe His 
    370         


<210>  4
<211>  22
<212>  DNA
<213>  Secuencia artificial

<220>
<223>  Cebador directo de PCR AP1, para amplificacin del extremo 5' del
       clon 3A22 de Anisakis simplex

<400>  4
ctaatacgac tcactatagg gc                                              22


<210>  5
<211>  28
<212>  DNA
<213>  Secuencia artificial

<220>
<223>  Cebador reverso de PCR SR1, para amplificacin del extremo 3' del
       clon 3A22 de Anisakis simplex

<400>  5
acccgcagta gttttatcca tttgttcg                                        28


<210>  6
<211>  26
<212>  DNA
<213>  Secuencia artificial

<220>
<223>  Cebador directo de PCR: SER5', para amplificacin del gen 
       completo de AniSerp

<400>  6
atgatgacag cattaccgtt tttaac                                          26


<210>  7
<211>  30
<212>  DNA
<213>  Secuencia artificial

<220>
<223>  Cebador inverso de PCR SER3', para la amplificacin del gen 
       completo de AniSerp

<400>  7
tcagtggaaa cgaccaataa acagaatgcg                                      30


<210>  8
<211>  27
<212>  DNA
<213>  Secuencia artificial

<220>
<223>  Cebador director de PCR 5'-SER-SPS

<400>  8
cccgggatgc agcagacaat cgatgat                                         27


<210>  9
<211>  28
<212>  DNA
<213>  Secuencia artificial

<220>
<223>  Cebador inverso de PCR 3'-SER-SPS

<400>  9
gcggccgctc agtggaaacg accaataa                                        28

