1、外源雌激素處理對(duì)唐魚卵黃蛋白原和雌激素受體基因表達(dá)的影響,張為民 中山大學(xué)生命科學(xué)學(xué)院,唐魚(Tanichthys albonubes),雄魚,雌魚,唐魚,唐魚為鯉科(Cyprinidae)魚丹亞科(Danioninae)唐魚屬(Tanichthys Lin, 1932) 僅分布于我國廣州白云山一帶山區(qū)小溪流中，瀕危物種，屬國家II級(jí)保護(hù)動(dòng)物 研究表明唐魚對(duì)重金屬Cu 2 + 等敏感,是一種較理想的環(huán)境重金屬污染指示生物,Question,唐魚對(duì)環(huán)境雌激素污染是否也敏感？,卵黃蛋白原,卵黃蛋白原(VTG)是卵生動(dòng)物中卵黃蛋白(yolk protein)的前體 ，由1100多個(gè)氨基酸組成 VTG

2、經(jīng)蛋白酶水解產(chǎn)生：卵黃脂磷蛋白I (lipovitellin I),卵黃高磷蛋白(phosvitin),和卵黃脂磷蛋白II ( lipovitellin II) 在魚類、兩棲類和鳥類中, VTG是在E2刺激下由雌性動(dòng)物肝臟中合成的 雄魚肝臟在E2刺激下也能合成VTG,雌激素受體,廣義的雌激素受體（estrogen receptor，ER）包括雌激素膜受體、胞質(zhì)受體和核受體 雌激素核受體介導(dǎo)雌激素的基因組效應(yīng) E2刺激VTG生物合成是由雌激素核受體介導(dǎo)的 環(huán)境雌激素也可作用在雌激素核受體上影響動(dòng)物內(nèi)分泌平衡,目的意義,克隆唐魚VTG和雌激素受體基因 分析E2對(duì)雄性唐魚這些基因表達(dá)的影響,方法,基

3、因克隆采用3 和5RACE方法 基因表達(dá)分析采用半定量RT-PCR方法,VTG: Nucleotide and deduced amino acid sequences,1 ACAAACCATCAGCAATGAGAGCTGTTGTGCTTGCCCCGACTGTAGCCCTTGTGGCGAGTC 1 M R A V V L A P T V A L V A S Q 61 AACAGATCAGCCTTGTTCCCGAGTTTGCCTCTGATAAGACCTATGTGTACAAGTATGAGG 17 Q I S L V P E F A S D K T Y V Y K Y E A 121 CTCTGCA

4、CTTGGGAGGACTTCCTCAAGAAGGTCTGGCTAGAGTCGGAGCAAAAGTCAACG 37 L H L G G L P Q E G L A R V G A K V N G 181 GCAAGGTTTACATCCGTGCCGAGACAGAGAGCACCTTTCTGATGAAGTTCTCAGATGTTC 57 K V Y I R A E T E S T F L M K F S D V Q 241 AACTCCATGAGTATACTGGCATTTGGCCCAGGGATCCTTTCGTTCCTGAAACTAAGCTCA 77 L H E Y T G I W P R D P F V

5、 P E T K L M 301 TGTCAACTATGACTGCTGAGCTGCAGATTCCCATTAAGTTTGAGTATACCAATGGTGTGG 97 S T M T A E L Q I P I K F E Y T N G V V 361 TTGGAAAGATATTCGCCCCTGCAGGAGTCTCCCCTATTGTTCTGAACTTTTACAGAGGAA 117 G K I F A P A G V S P I V L N F Y R G I 421 TCCTCAACATCTTTCAGCTTAACCTCAAGAAGACCCAGAACATCTATGAGCTGCAAGAGG 137 L

6、 N I F Q L N L K K T Q N I Y E L Q E A 481 CTGGAGTTCAGGGAGTGTGCAGGACCCACTATGTCATCAGTGAGGATTCGAAAGCCAACC 157 G V Q G V C R T H Y V I S E D S K A N H 541 ACATTACTGTCACCAAGTCTAAGGATCTGAGCCACTGCCAGGAGAGAATCACAAAAGATT 177 I T V T K S K D L S H C Q E R I T K D F 601 TCGGCTTGGCATACACTGAGAAGTGTGACGAATGCACTG

7、AGCGGGTAAAGAGTCTGATTG 197 G L A Y T E K C D E C T E R V K S L I E 661 AAACCGCATCATACAACTACATCATGAAACCAACATCTGCCGGTGTATTGGTGTCTGAAG 217 T A S Y N Y I M K P T S A G V L V S E A 721 CAACAGTTGAGGAAGTGTATCAGTTTTCACCATTCAATGAGATCCATGGTGCTGCCCAGA 237 T V E E V Y Q F S P F N E I H G A A Q M 781 TGGAAGTAAAGC

8、AATCTTTGGCTTTTGTTGAGATTGAGAAGACCCCTATTGCTCCATTTA 257 E V K Q S L A F V E I E K T P I A P F K 841 AAGCTGATTACTTGGCTCGTGGATCTCTGCGGTATGAGTTTGCAAGTGAGATTCTTCAGG 277 A D Y L A R G S L R Y E F A S E I L Q A 901 CCCCCATTCAACTCATGAAGATCAGTGATGCACCAGCCCAGATTACTGAGATCCTGAAGC 297 P I Q L M K I S D A P A Q I T

9、 E I L K H 961 ACTTGGTTGAAAACAATGTGGCCATGGTCCATGATGATGCTCCACTTAAGTTTGTTCAGC 317 L V E N N V A M V H D D A P L K F V Q L 1021 TCGTCCAGCTCATGCGTGCTGCAACCTTTGAGGATATTGAGTCCATCTGGAAGCAGTGCA 337 V Q L M R A A T F E D I E S I W K Q C K,1081 AGGACAAACCAGTTTACAGGCGTTGGTTTCTGGATGCTCTTCCTGCTGTTGGTACACCAG 357

10、D K P V Y R R W F L D A L P A V G T P V 1141 TCATTGTAAAATTCATCAAGGAGAAGTTCCTGGATGGTGAACTTTCCATTCCTGAGTTCA 377 I V K F I K E K F L D G E L S I P E F I 1201 TTCAGGCTCTTGTAGTTGGTTTGCAAATGGTTTCTGCTGATTTGGACACCATCGAGTTGA 397 Q A L V V G L Q M V S A D L D T I E L T 1261 CAGCTAGTTTGGTTAAGCACGATAAAATCGCCACA

11、ATCCCAGCTCTTCGTGAAGTCGTCA 417 A S L V K H D K I A T I P A L R E V V M 1321 TGCTTGGATATGGTTCCATGATTGCCAAATACTGCGTTGAGGTTCCTTCTTGCCCTGCTG 437 L G Y G S M I A K Y C V E V P S C P A D 1381 ACCTCCTCAGGCCCATCCATGATCTTGCTGCAGAGGCAATTTCCAAGAATGACATTCCTG 457 L L R P I H D L A A E A I S K N D I P E 1441 AAATC

12、ACGTTGGCTCTGAAAGTTCTGGGCAATGCTGGTCATCCTGCTAGTCTTAAACCCA 477 I T L A L K V L G N A G H P A S L K P I 1501 TCATGAAGCTTCTGCCAGGATTGAGAACTTCAGCTACTTCTTTGCCTCTTAGAGTCCAGG 497 M K L L P G L R T S A T S L P L R V Q V 1561 TTGATGCCATCTTGGCCCTGAGGAACATTGCCAAAAAAGAACACAAACTGGTTCAGCCAG 517 D A I L A L R N I A

13、K K E H K L V Q P V 1621 TGGCCATGCAGATTGTTTTGGACAGGGCTCTCCACCCTGAAGTGCGTATGGTTGCTTGTG 537 A M Q I V L D R A L H P E V R M V A C V 1681 TTGTGTTGTTCGAGGCCAAGCCCTCAGTGGCTCTCGTCTCCAATCTTGCTGGTGCCATGA 557 V L F E A K P S V A L V S N L A G A M K 1741 AGACTGAGACTAACATGCATGTCGCAAGCTTTGCTTATTCCCACATCAAGTCATT

14、GACCA 577 T E T N M H V A S F A Y S H I K S L T R 1801 GAATCAGTGCTCCTGATATGGCATCCGTTGCTGGTGCTGCTAATGTTGCCGTCAAGCTCA 597 I S A P D M A S V A G A A N V A V K L M 1861 TGAGCCGCAAACTGGACAGGCTTAGCTTTCATTTCAGCAGAGCCCATCAGCTGGACTACT 617 S R K L D R L S F H F S R A H Q L D Y Y 1921 ATCATACTCCTCTTATGATTGGAGC

15、TGCTGGCAGTGCCTACATGATCAATGATGCCGCCA 637 H T P L M I G A A G S A Y M I N D A A T 1981 CCATCCTGCCCAGAGCTGTTGTAGCTAAAACCCGTGCTTACCTGGCTGGAGCTGCTGCTG 657 I L P R A V V A K T R A Y L A G A A A D 2041 ATGTTCTTGAGATCGGTGTGAGAACTGAGGGAATCCAGGAGGCTCTTCTGAAATCTTCTG 677 V L E I G V R T E G I Q E A L L K S S A

16、2101 CTGCAGATCAAAGTGTTGACCGTATGACAAAGATTAAGCGCACACTGAGAGCTCTTACAA 697 A D Q S V D R M T K I K R T L R A L T N,(4171bp，1326aa, full-length ),3121 ACAGTAACAGTAACAGCAGTAGCAGTAGCAGCAGCAGCCGTCGCAGCCGCAGTAACGGTA 1037 S N S N S S S S S S S S R R S R S N G S 3181 GTAGCAGCAGCAGCAGCAGAAGCTCAAGCTCAAGCTCAAGCTCA

17、AGCTCCTCCATGTCCA 1057 S S S S S R S S S S S S S S S S S M S S 3241 GCTCTCGTATGAATAAGGCTACCACTGTAATGAAGCCATTCAAGAAGTTCCACAAAGACC 1077 S R M N K A T T V M K P F K K F H K D Q 3301 AGTACTTGAATGCTCATGGAAATTCAAAGGCAGTAAGCAGCGGAGGCTCAGAATCTAGCT 1097 Y L N A H G N S K A V S S G G S E S S F 3361 TTCAGCAAATC

18、CAGAAACAGAACAAGTTCCTCGGAAATGCTATTCCACCTATTTTTGCCG 1117 Q Q I Q K Q N K F L G N A I P P I F A V 3421 TCATTGCTCGCGCTGTGAGAGTTGACCAAAAGCTGCTGGGCTACCAACTTGCTGCTTACT 1137 I A R A V R V D Q K L L G Y Q L A A Y F 3481 TTGACAAGTCAACATCAAGAGTTCAGTTCATCATTGCCTCCATTGCTGAAGATGACAACC 1157 D K S T S R V Q F I I A

19、 S I A E D D N L 3541 TGAAGATCTGTGCTGATGGTGCCCTGCTGAGCCAGCTCAAAGTCACTGCCAAGGTTGCTT 1177 K I C A D G A L L S Q L K V T A K V A W 3601 GGGGTCAGGAATGCCAACAGTATGCCATCACTGCTAAAGCTGAAACCGGTGTCCTGGATG 1197 G Q E C Q Q Y A I T A K A E T G V L D E 3661 AATTTCCTGCAGCACGCCTGGAGCTGGAATGGGAGAGGCTGCCAGAAGTTGTCTAC

20、AAAT 1217 F P A A R L E L E W E R L P E V V Y K Y 3721 ACGCCAAAAAGCTGTCTAAGCACATCCCTATGGCAGCTTTTCAGACTGGCCTTAGACTTG 1237 A K K L S K H I P M A A F Q T G L R L E 3781 AAAAGGCAAAGAACTGTGAGAAAGAGATTGAACTGACTGCAGCCTTGCCAACTCAGAGAT 1257 K A K N C E K E I E L T A A L P T Q R S 3841 CCTTGAATGTCATCGCTAGGATT

21、CCAGAGATGACACTGTCAAGGATGGATATTCCCCTCC 1277 L N V I A R I P E M T L S R M D I P L P 3901 CTGTCGCTGTTCCCATCAATCCAGATGGAACTTTTTCCATTCATTACGACGATGACGTTC 1297 V A V P I N P D G T F S I H Y D D D V L 3961 TCTCCTGGATCGAGAAACAAATCAATGAAGCATAAGACGTGAAGCATTAAATTTTACACT 1317 S W I E K Q I N E A * 4021 TTTCAGAA

22、GAAAATGAGAGTGGTGTAAAATTGATCATGCGGAATTGATTGTTTCTCTGAC 4081 AGTTTAATTGAACTGTACCACTGTTGTTATTCAAGTTTAACACTGATAAACACCCTGCAA 4141 GTTAAAAAAAAAAAAAAAAAAAAAAAAAAAA,2161 ACTGGAAAGCTCTGCCAACCAATCAGCCATTGGCTTCAGCATATGTAAAATTGTTTGGGC 717 W K A L P T N Q P L A S A Y V K L F G Q 2221 AGGAAGTTGCATTCGCTAATATTGACAGG

23、ACCATCATTGAGCAAGCAATACCGATTGTGA 737 E V A F A N I D R T I I E Q A I P I V T 2281 CTGGACCAAAACCACGTGAACTGTTGAAGGATGCCCTTAAAGCTTTGCAGGAAGGATTTT 757 G P K P R E L L K D A L K A L Q E G F S 2341 CTTACCAATATGCCAAACCCCTGCTTGTAGCTGAAGTGCGCCGTGTCTTGCCCACATCAG 777 Y Q Y A K P L L V A E V R R V L P T S V 2401

24、 TTGGTTTTCCAATGGAGCTCAGTGTTTACTCTGTTGTTGTTGGTGCTGCAAGTCTCAATG 797 G F P M E L S V Y S V V V G A A S L N V 2461 TTAAGGCCACCATTACACCTCCTCTCCCTGAGAAGATTGAGACTATGACTCTTGAACAGC 817 K A T I T P P L P E K I E T M T L E Q L 2521 TCAAGAAGACTGATGTTCAATTCGAAGCTGAAGCTAGACCAAGTATTGCCATACAGAAAT 837 K K T D V Q F

25、E A E A R P S I A I Q K F 2581 TTGCTGTGATGGGAGTGAACACTGCCTTTATCCAAGCTGCTGTTATGGCAAAGGGAGAAA 857 A V M G V N T A F I Q A A V M A K G E I 2641 TCCGCATGATTGCTCCAGGAAAAGTGGCTGCTAGAGCAGACTTACTCAAGGGCAACTACA 877 R M I A P G K V A A R A D L L K G N Y K 2701 AGGTTGAAGCTTTGCCTGTTGAGCTTCCTGAGCACATTGCTGATATGAG

26、CTTTGAGACGT 897 V E A L P V E L P E H I A D M S F E T F 2761 TTGCTGTGATTAGAAACGTTGAAGAGCCCAATGCTGATAGGATTGTTCCCTTAGTACCTG A V I R N V E E P N A D R I V P L V P E 2821 AGATGGCCATGCAAAATTTCCAGAGTCGTTCTGACTATTCGTCTTCTGAAGATGATTCAG 937 M A M Q N F Q S R S D Y S S S E D D S D 2881 ACGATACTGCTGTGAGAGCTTCT

27、GCTCCATATCACGAGACTCTCTGTTATGCTGTCCCAT 957 D T A V R A S A P Y H E T L C Y A V P Y 2941 ACATTGAAATCAAGGGATGTGTTGAGGTGCACTCTCAGAATGCTGCCCATATCAGAAATA 977 I E I K G C V E V H S Q N A A H I R N T 3001 CTTCTTTCTTCCACATAATTGGACAGCACTCAGCCCGTGCTGCAGTTTCAAGAACTGAAG 997 S F F H I I G Q H S A R A A V S R T E

28、G 3061 GTCCTGCAGTTGAAAGACTGGAGACTGAAGATAAAAATCGACGTGTCTCTTCTGAAAGCA 1017 P A V E R L E T E D K N R R V S S E S N 1,卵黃脂磷蛋白 I,卵黃脂磷蛋白 II,卵黃高磷蛋白,卵黃高磷蛋白,VTG: Nucleotide and deduced amino acid sequences,VTG phylogenetic tree,ER: Nucleotide and deduced amino acid sequences,1 GTCTCTGTCGGATACTACCCGGCTCCCACGG

29、ACCCCCACGCACCCCCCGCCGAAGAACAT V S V G Y Y P A P T D P H A P P A E E H 61 CTGCAGACGCTAGGCGCCGGATCCAGTAGCCCTCTCATGTTTGCACCGTCCAGTCCTCAG L Q T L G A G S S S P L M F A P S S P Q 121 CTGTCCCCGTACCTCAACCATCATGGAGGACAGCACTCGACCCAGCAGGTGTCCTACTAC L S P Y L N H H G G Q H S T Q Q V S Y Y 181 CTGGACACTTCTTCTAG

30、CACAGTGTACAGGTCCAGTGTGGTCTCCTCTCAGCAGGCAGGT L D T S S S T V Y R S S V V S S Q Q A G 241 GTGGGTCTGTGTGAGGTGTTGTGCAGTGCGACTGACAGGCAGGAGTTGTACAGCGGATCC V G L C E V L C S A T D R Q E L Y S G S 301 AGAGCTGCAGGAGGGTTTGATTCAGAGAAGGAGACGCGTTTCTGTGCGGTGTGCAGTGAC R A A G G F D S E K E T R F C A V C S D 361 TA

31、TGCCTCGGGCTATCATTATGGAGTCTGGTCCTGTGAGGGATGCAAAGCTTTCTTCAAG Y A S G Y H Y G V W S C E G C K A F F K 421 AGAAGTATACAAGGTCACAATGACTATGTTTGTCCAGCAACCAACCAGTGCACTATTGAC R S I Q G H N D Y V C P A T N Q C T I D 481 AGAAACCGCAGGAAGAGCTGCCAAGCATGCAGACTACGCAAGTGCTATGAAGTAGGCATG R N R R K S C Q A C R L R K C Y

32、 E V G M 541 ATGAAAGGAGGTATTCGCAAGGACCGCGGCGGCCGCACTATCAGACGTGAGCGCCCAAGG M K G G I R K D R G G R T I R R E R P R 601 AGCAGCATCGAGGAGAAGAGCTACAGTGAGCAGTCAAGCCGTGCCGGACTGAGGACAGCT S S I E E K S Y S E Q S S R A G L R T A 661 CAGGACAAGAGAAAGAAGAGCAGCAGTGGCGGGATGGTCAGTGCTTTATGTATGCCACCT Q D K R K K S S

33、S G G M V S A L C M P P 721 GACCAGGTGCTGATGTTGCTTCTGGGCGCAGAGCCGCCGGCTGTCTGTTCACGTCAGAAA D Q V L M L L L G A E P P A V C S R Q K 781 CACAGTCCCCCGTACACCGAGATCACCATGATGTCCCTGCTCACAAACATGGCTGACAAG H S P P Y T E I T M M S L L T N M A D K,841 GAACTCGTCCACATGATCGCCTGGGCTAAGAAAGTACCAGGGTTCCAGGACCTCTCTCTG E

34、 L V H M I A W A K K V P G F Q D L S L 901 CATGACCAGGTTCAGCTGTTGGAGAGCTCTTGGCTGGAGGTGTTGATGATCGGCCTCATT H D Q V Q L L E S S W L E V L M I G L I 961 TGGAGGTCCATTCATTCACCTGGAAAACTCATCTTTGCTCAAGATCTCATCCTCGATAGG W R S I H S P G K L I F A Q D L I L D R 1021 AGTGAAGGAGAATGTGTCGAGGGGATGGCTGAGATTTTCGACATGC

35、TTTTGGCGACTGTG S E G E C V E G M A E I F D M L L A T V 1081 GCTCGATTCCGCAGTCTCAAACTCAAGCTGGAGGAATTTGTATGTCTTAAAGCCATCATA A R F R S L K L K L E E F V C L K A I I 1141 CTTCTCAATTCTGGTGCATTTTCATTCTGCTCCAGTCCAGTGGAGCCCCTGATGGACAGC L L N S G A F S F C S S P V E P L M D S 1201 TTCATGGTCCAGTGTATGCTGGACAACA

36、TCACTGATGCCCTCATTCACGGCATCAGTAAA F M V Q C M L D N I T D A L I H G I S K 1261 TCAGGTGCCTCTCTGCAGCTGCAGTCCCGCCGTCAGGCCCAGCTCCTTCTGCTGCTGTCC S G A S L Q L Q S R R Q A Q L L L L L S 1321 CACATCAGACACATGAGCAACAAAGGAATGGAGCACTTATACCATATGAAATGTAAGAAT H I R H M S N K G M E H L Y H M K C K N 1381 CGAGTCCCGC

37、TGTATGATCTCTTGCTGGAGATGCTGGATGCCCAGAGATTCCAGTCACCA R V P L Y D L L L E M L D A Q R F Q S P 1441 GGAAAGGTGCAGCAACTGTGGGCAGAGGGTGAGAAAGAGCCCCTGTTTACACCCACAACC G K V Q Q L W A E G E K E P L F T P T T 1501 AGCAGCAGCAGCAACAGCCCCGGAGCCATGCAGCCCAACACTGCCTTTCTTAGCCAAAGT S S S S N S P G A M Q P N T A F L S Q

38、 S 1561 CCAGATACCTAATCTATATCCTGACCCGGTACCACCTACAATCCCAACAGCTGCAGAATG P D T 1621 TGATTACGAACTAATAATAAGGACTGTGATTCAAAACATTGCTGACTAATAGTTTTTGAA 1681 GGGGAGTCTCTCCTTGTGCCTTATTAGAGTCCACCTTAGATTCCTATATAGAAATTAACA 1741 ATTAAATTAACCTATTTTTAGAAGCACACACACATGCTAAGTCTTAAATTTGTAACAAGG 1801 CTTAAGATTCAAAACAGCTACA

39、GTTTGTAAGTGTGTTATAT,(1841 bp, 491 aa , partial),ER phylogenetic tree,ER: Nucleotide and deduced amino acid sequences,TG 3 GACTCTGGCAAGGCTGATCGAGGGGACTCGCCTAACTTCTTACCCCGTCTGTACTCTTCC D S G K A D R G D S P N F L P R L Y S S 61 CCACTCGGCATGGACAACCAGACCATCTGCATTCCATCTCCGTATTTGGAAGCCTGTCAG P L G M D N Q

40、 T I C I P S P Y L E A C Q 121 GATTACTCACCACCGCACGGGGGAGAGTTCAACCACGGAGCTTTAACACTCTACAGCCCG D Y S P P H G G E F N H G A L T L Y S P 181 GTGTCCTCCTCAGTGCTAGGGTACCCTCATCCCCCCGTGTCTGAAAGCCTGGTCCCGCTC V S S S V L G Y P H P P V S E S L V P L 241 AGTCCCACAATCTACTGGCCTTCTCACACGGCACACACTGCGCTTTCTCTGCACTGCCC

41、G S P T I Y W P S H T A H T A L S L H C P 301 CCTCCTCTGGCCTACAGCGAAACACACTCACACACTGCCTGGGAGGAAGCCAAGACACAC P P L A Y S E T H S H T A W E E A K T H 361 ACGCTTAACCACAGCAGTTCTGTCCTTACTCATGCAAAGCAGTTAGGGCTGCAAGTGGAA T L N H S S S V L T H A K Q L G L Q V E 421 GGTGACGATGGCTTAAATCCTTCTCCAGGCATTGTGGGTAAAGG

42、AGACACACACTTCTGT G D D G L N P S P G I V G K G D T H F C 481 GCGGTGTGTCATGACTACGCCTCTGGGTATCACTATGGTGTCTGGTCATGTGAGGGGTGC A V C H D Y A S G Y H Y G V W S C E G C 541 AAGGCTTTCTTCAAACGGAGCATTCAAGGACACAATGACTATATTTGTCCAGCCACCAAC K A F F K R S I Q G H N D Y I C P A T N 601 CAGTGCACCATCGACAAGAGCCGACGCAA

43、GAGCTGTCAGGCCTGTCGACTCCGCAAGTGC Q C T I D K S R R K S C Q A C R L R K C 661 TACGAGATGGGCATGATGAAGTGTGGTGTGAGGCGTGAACGCTGCAGTTACCGTGGTGCT Y E M G M M K C G V R R E R C S Y R G A 721 CGCCATCGGCGCAACCCCCAAATCCGAGATGGCTCGGGCGGGGCGGGGGCGTTAGGGGTC R H R R N P Q I R D G S G G A G A L G V 781 AGAGGTTGTTCCCA

44、GCATCAGTTAGAAATCCCTCTCAATCCCACTGATCACCTCTTCCCT R G C S Q H Q L E I P L N P T D H L F P 841 TCAGGGAGCAGAGCTGAGGGATCTGGCCTGAGCTTCTCTCCTGAGCAGCTGGTCAACTGC S G S R A E G S G L S F S P E Q L V N C 901 ATTCTGGAGGCGGAGCCTCCTCAGATTTGCCTGAGAGAGCCAGTGACAAAGCCGTACACG I L E A E P P Q I C L R E P V T K P Y T 961

45、 GAGGCCAGCATGATGATGTCACTCACCACCCTCGCTGACAAGGAGCTGGTGCTCATGATC E A S M M M S L T T L A D K E L V L M I 1021 AGCTGGGCCAAGAAGATACCAGGATTCGTAGAGCTGACACTTTCGGATCAGGTGCATCTA S W A K K I P G F V E L T L S D Q V H L,1081 TTGGAATGCTGTTGGCTGGATATTCCGATGTTGGGATTGATGTGGAGATCTGTGGATCAT L E C C W L D I P M L G L

46、M W R S V D H 1141 CCTGGGGAAACTCATCTTTTCACCCGACCTCAACTGAACAGGGATGAATGGAATTGTGTT P G E T H L F T R P Q L N R D E W N C V 1201 GAAGGCTTAATGGAGATCTTTGACATGCTGTTAGCAACCACCTCTAGATTCAGAGAGCTG E G L M E I F D M L L A T T S R F R E L 1261 AAGCTACAGAGAGAGGAATACGTCTGTCTCAAAGCCATGATTCTTCTCAACTCCAACAAC K L Q R

47、E E Y V C L K A M I L L N S N N 1321 TGTTCAAGCTTGTCACAGACTCCTGAAGATGTGGAGAGCCGTGGAAAGGTTCTGAGGCTG C S S L S Q T P E D V E S R G K V L R L 1381 CTGGACTCTGTGACTGACGCTCTGGTGTGGAGCATCTCCAGAACGGGCTTGTCCTCTCAG L D S V T D A L V W S I S R T G L S S Q 1441 CAGCAGTCCACCCGACTCGCCCATCTGCTGATGCTGCTCTCACATATTCGA

48、CACCTCAGC Q Q S T R L A H L L M L L S H I R H L S 1501 AACAAAGGCATCGAGCATCTGTCAAGCATGAAGAGGAAAAACGTGGTGCTGCTGTACGAT N K G I E H L S S M K R K N V V L L Y D 1561 CTTCTGCTGGAGATGCTGGATGCCAACACGTCCCAAAGCAGCAGGATGCTGGCTGATCAC L L L E M L D A N T S Q S S R M L A D H 1621 TCAGAAACCTCTCTCCGGCCGGACACAGAACAGACCCCAGAGGTCCTCTACACATCCAGA S E T S L R P D T E Q T P E V L Y T S R 1681 GCAAGAGAGAGCGACCCGGAGACCCGGCACAGTCCGCACGCTGAGGAGACATTGCACTCT A R E S D P E T R H S P H A E E T L H S 1741 TGTCTTCATCGAGAGGACATGGACACAGACTGACAGTCTTTCTTCTGTATGGTAACTGAA C L H R E D M D T D 1801 CTAGG