PH N LËP Vµ X C ÞNH TR NH Tù GIEN M HãA PR«TªIN Vá CñA VIRót Y ë KHOAI T Y TRåNG T¹I TH I NGUY N - PDF

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32(1): T¹p chý Sinh häc PH N LËP Vµ X C ÞNH TR NH Tù GIEN M HãA PR«TªIN Vá CñA VIRót Y ë KHOAI T Y TRåNG T¹I TH I NGUY N NGUYÔN THÞ T M Tr êng ¹i häc S ph¹m, ¹i häc Th i Nguyªn CHU HOµNG MËU

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32(1): T¹p chý Sinh häc PH N LËP Vµ X C ÞNH TR NH Tù GIEN M HãA PR«TªIN Vá CñA VIRót Y ë KHOAI T Y TRåNG T¹I TH I NGUY N NGUYÔN THÞ T M Tr êng ¹i häc S ph¹m, ¹i häc Th i Nguyªn CHU HOµNG MËU ¹i häc Th i Nguyªn NGUYÔN Vò THANH THANH Tr êng ¹i häc Khoa häc, ¹i häc Th i Nguyªn Khoai t y (Solanum tuberosum L.) lµ c y l ng thùc îc trång réng r i trªn thõ giíi. Virót X (Potato virus X - PVX), virót Y (Potato virus Y - PVY), virót g y xo n l (Potato leaf roll virus - PLRV), virót A (Potato virus A - PVA), virót M (Potato virus M - PVM), virót S (Potato virus S - PVS)... g y ra c c bönh xo¾n lïn, kh m l vµ cuèn l lµ c c virót g y bönh trªn khoai t y ë ViÖt Nam. V thõ c y sinh tr ëng chëm, sè l îng vµ khèi l îng cñ bþ gi m ng kó, do vëy n ng suêt bþ gi m râ röt. ë ViÖt Nam, bönh virót cã ë kh¾p c c vïng trång khoai t y, phæ biõn g y h¹i næng lµ virót X vµ Y [1, 2]. PVY lµ Potyvirus thuéc hä Potyviridae víi hö gien chøa ARN sîi n, kých th íc kho ng 9,7kb. PVY g y bönh nghiªm träng trªn khoai t y, thuèc l, cµ chua vµ mét sè c y trång thuéc hä Cµ. PVY cã 5 nhãm kh c nhau, ã lµ: PVY O, PVY N, PVY NTN, PVY N Wi vµ PVY C. PVY gåm c c protein sau: P1, HC (Helper Component), P3, CI (Cylindrical Inclusion), Nia (Nuclear Inclusion A), Nib (Nuclear Inclusion B), CP (Coat Protein), 6K1, 6K2 [3]. Trong bµi b o nµy, chóng t«i nghiªn cøu ph n lëp vµ x c Þnh tr nh tù gien m hãa protein vá (CP-Coat Protein) cña virót PVY ë khoai t y îc trång t¹i Th i Nguyªn nh»m phôc vô cho viöc t¹o gièng khoai t y chuyón gien kh ng l¹i c c virót c«ng bè. Tr nh tù nucleotit vïng m hãa cña gien CP tõ PVY cña khoai t y trång t¹i Th i Nguyªn Òu dµi 801 nucleotit, m hãa 267 axit amin. Tr nh tù nucleotit cña gien CP vµ tr nh tù axit amin cña protein CP tõ PVY cña khoai t y ph n lëp Òu t ng ång cao víi tr nh tù c«ng bè trªn Ng n hµng gien NCBI. 1. VËt liöu I. PH NG PH P NGHI N CøU Hai méu l khoai t y bþ nhiôm bönh thu tõ huyön Phó B nh tønh Th i Nguyªn îc ký hiöu ThaiNguyen1 vµ ThaiNguyen2. C c lo¹i hãa chêt, dông cô vµ thiõt bþ phôc vô cho thý nghiöm sinh häc ph n tö. 2. Ph ng ph p ARN tæng sè îc t ch chiõt theo h íng dén sö dông hãa chêt Trizol Reagent cña h ng Invitrogen (Mü). Tæng hîp cadn theo bé kit First stand cdna synthesis cña h ng Fermentas. Nh n gien CP b»ng kü thuët PCR. PCR îc tiõn hµnh víi tæng thó tých ph n øng 25 µl gåm: cadn (50 ng/µl) 3 µl, måi (10 µm) 2 µl, dntp (2,5 mm) 2 µl, MgCl 2 (25 mm) 2,5 µl, Taq polymeraza (5 unit/µl) 0,5 µl, dung dþch Öm PCR (10X) 2,5 µl, H 2 O khö ion 12,5 µl. Chu tr nh nhiöt bao gåm c c b íc sau: 94 o C-3 phót; 94 o C-1 phót, 52 o C-50 gi y, 72 o C-1 phót 30 gi y læp l¹i 30 chu k ; 72 o C - 10 phót vµ l u gi ë 4 o C. S n phèm PCR nh n gien CP îc kióm tra b»ng iön di trªn gel agaroza 1%. Gien îc lµm s¹ch (th«i gel) theo bé Kit cña h ng Bioneer vµ g¾n vµo vðct pbt (phßng C«ng nghö tõ bµo thùc vët - ViÖn C«ng nghö Sinh häc cung cêp), sau ã îc biõn n¹p vµo tõ bµo kh biõn E. coli 81 chñng DH5α. T ch plasmit theo bé Kit cña h ng Bioneer. Tr nh tù nucleotit cña gien CP îc x c Þnh trªn m y äc tr nh tù nucleotit tù éng ABI 3100 Advant Genetic Analyzer cña h ng Ampplied Biosystem. KÕt qu äc tr nh tù gien îc xö lý b»ng phçn mòm DNAstar vµ BioEdit. II. KÕT QU Vµ TH O LUËN 1. KÕt qu nh n gien CP Ó ph n lëp gien CP, chóng t«i thu thëp méu l khoai t y bþ nhiôm virót tõ huyön Phó B nh, tønh Th i Nguyªn. Sau ã, chóng t«i tiõn hµnh t ch ARN tæng sè tõ méu l nhiôm bönh vµ tæng hîp cadn tõ ARN tæng sè. Ó ph t hiön sù cã mæt cña gien CP ë hai méu khoai t y bþ nhiôm bönh, chóng t«i sö dông cæp måi Æc hiöu îc cung cêp bëi Phßng C«ng nghö tõ bµo thùc vët-viön C«ng nghö sinh häc Ó nh n gien CP. o¹n gien CP îc nh n lªn b»ng ph ng ph p PCR, kõt qu nh n gien, clony-pcr îc kióm tra b»ng c ch iön di trªn gel agaroza 1% vµ îc thó hiön trªn h nh 1. M 1 2 M 1200bp 1500 bp 1000 bp 1324bp a H nh 1. KÕt qu PCR nh n gien CP vµ clony-pcr Ghi chó: M. ChØ thþ ph n tö 1kb; 1. ThaiNguyen1; 2. ThaiNguyen2. b H nh 1a cho thêy, o¹n gien îc nh n lªn cã kých th íc kho ng 1200 bp. Ó x c Þnh tr nh tù gien CP, chóng t«i tiõn hµnh g¾n s n phèm PCR tinh s¹ch ë trªn vµo vðct t ch dßng, biõn n¹p vµo vi khuèn E.coli vµ chän dßng gien. Qu tr nh t ch dßng îc thùc hiön b»ng c ch g¾n s n phèm PCR tinh s¹ch vµo vðct t ch dßng pbt, biõn n¹p vµo tõ bµo kh biõn chñng E. coli DH5α vµ îc cêy tr i trªn Üa petri cã m«i tr êng LB Æc bæ sung ampixillin 100 mg/ml, X-gal 40 mg/ml vµ IPTG 100 µm. ñ c c Üa petri cêy tr i ë 37 o C trong 16 giê, kõt qu thu îc c khuèn l¹c xanh vµ tr¾ng. Chän khuèn l¹c tr¾ng nu«i trong m«i tr êng LB láng cã bæ sung ampixillin 100 mg/ml qua ªm. LÊy khuèn cña mçi méu ch¹y ph n øng clony PCR víi cæp måi puc18 Ó x c Þnh khuèn l¹c cã plasmit mang gien mong muèn. V cæp måi puc18 lµ cæp måi îc thiõt kõ chung cho c c vðct t¹o dßng nªn khi kióm tra s n phèm PCR võa dßng ho th kých th íc cña c c o¹n gien võa nh n lªn sï nhiòu h n kho ng 124 nucleotit so víi nh n b»ng cæp måi Æc hiöu. M 1 2 H nh 2. KÕt qu iön di t ch plasmýt mang gien CP S n phèm clony PCR tõ nh ng khuèn l¹c 82 tr¾ng Òu cho kõt qu d ng týnh. TiÕn hµnh chän khuèn l¹c tr¾ng t ng øng víi 2 méu nghiªn cøu cã s n phèm clony PCR ë trªn nu«i trong m«i tr êng LB láng sau ã t ch plasmit theo bé kit cña h ng Bioneer. S n phèm ADN plasmit îc iön di trªn gel agaroza 1%, kõt qu îc thó hiön ë h nh 2. KÕt qu iön di trªn h nh 2 cho thêy, s n phèm t ch plasmit s¹ch, m b o chêt l îng vµ sè l îng Ó tiõn hµnh äc tr nh tù nucleotit cña gien CP. Ó x c Þnh chýnh x c tr nh tù nucleotit cña gien CP t ch dßng, chóng t«i tiõn hµnh äc tr nh tù nucleotit cña gien CP trªn m y äc tù éng ABI 3100 Avant Genetic Analyzer theo hai chiòu xu«i vµ ng îc. KÕt qu cho thêy, chiòu dµi gien CP ë 2 méu nghiªn cøu Òu cã kých th íc 1201 nucleotit. Khi so s nh 2 tr nh tù nµy trong BLAST cña NCBI, kõt qu cho biõt y lµ c c tr nh tù gien m ho CP cña virót PVY ë khoai t y. Chóng t«i kõt luën nh n, t ch dßng vµ äc tr nh tù thµnh c«ng o¹n gien m ho CP cña virót PVY ë khoai t y. 2. So s nh tr nh tù gien vïng m hãa axit amin cña hai méu nghiªn cøu vµ so s nh tr nh tù axit amin cña protein CP ë mét sè tr nh tù ng ký trªn NCBI Chóng t«i quan t m tíi tr nh tù vïng m hãa cña gien CP virót PVY ë hai méu nghiªn cøu. Vïng m hãa axit amin cña gien CP virót PVY ë 2 méu ThaiNguyen1 vµ ThaiNguyen2 Òu cã chiòu dµi 801 nucleotit (h nh 3) ThaiNguyen1 GCAAATGACA CAATTGATGC AGGAGAAAGC AACAAGAAAG ATGCAAAACC ThaiNguyen2 GCAAATGACA CAATCGATGC AGGAGGAAGC AACAAGAAGG ATGCAAAACA ThaiNguyen1 AGAGCAAGGC AGCATCCAGT CAAACCTGAA CAAAGGAAAA GATAAGGATG ThaiNguyen2 AGAGCAAGGC AGCATCCAGC CAAACCTGAA CAAAGAAAAA GATAAGGACG ThaiNguyen1 TGAATGCTGG TACATCTGGG ACACATACTG TGCCGAGAAT CAAGGCTATC ThaiNguyen2 TGAATGCTGG AACATCTGGG ACACATACTG TGCCGAGAAT CAAGGCTATC ThaiNguyen1 ACGTCCAAAA TGAGAATGCC CAAAAGCAAG GGAGCAACCG TGCTAAATTT ThaiNguyen2 ACGTCCAAAA TGAGAATGCC CAAAAGCAAG GGAGCAACCG TGCTAAATTT ThaiNguyen1 AGAACACTTG CTTGAGTACG CTCCACAACA AATTGATATT TCAAATACTC ThaiNguyen2 AGAACACTTG CTTGAGTACG CTCCACAACA AATTGATATT TCAAATACTC ThaiNguyen1 GGGCAACTCA ATCACAGTTT GATGCGTGGT ATGAGGCAGT GCGGATGGCA ThaiNguyen2 GGGCAACTCA ATCACAGTTT GATGCGTGGT ATGAGGCAGT GCGGATGGCA ThaiNguyen1 TACGACATAG GAGAAACTGA GATGCCAACT GTGATGAATG GGCTTATGGT ThaiNguyen2 TACGACATAG GAGAAACTGA GATGCCAACT GTGATGAATG GGCTTATGGT ThaiNguyen1 TTGGTGCATT GAAAATGGAA CCTCGCCAAA TGTCAACGGA GTTTGGGTTA ThaiNguyen2 TTGGTGCATT GAAAATGGAA CCTCGCCAAA TGTCAACGGA GTTTGGGTTA ThaiNguyen1 TGATGGATGA GAATGAACAA GTTGAGTACC CGTTGAAACC AATCGTTGAG ThaiNguyen2 TGATGGATGA GAATGAACAA GTTGAGTACC CGTTGAAACC AATCGTTGAG ThaiNguyen1 AATGCAAAAC CAACCCTTAG GCAAATCATG GCACATTTCT CAGATGTTGC 83 ThaiNguyen2 AATGCAAAAC CAACCCTTAG GCAAATCATG GCACATTTCT CAGATGTTGC ThaiNguyen1 AGAAGCGTAT ATAGAAATGC GCAACAAAAA GGAACCATAT ATGCCACGAT ThaiNguyen2 AGAAGCGTAT ATAGAAATGC GCAACAAAAA GGAACCATAT ATGCCACGAT ThaiNguyen1 ATGGTTTAAT TCGAAATCTG CGGGATGTGG GTTTAGCGCG TTATGCCTTT ThaiNguyen2 ATGGTTTAAT TCGAAATCTG CGGGATGTGG GTTTAGCGCG TTATGCCTTT ThaiNguyen1 GACTTTTATG AAGTCACATC ACGAACACCA GTGAGGGCTA GGGAAGCGCA ThaiNguyen2 GACTTTTATG AAGTCACATC ACGAACACCA GTGAGGGCTA GGGAAGCGCA ThaiNguyen1 CATTCAAATG AAGGCCGCAG CATTGAAATC AGCCCAACCT CGACTTTTCG ThaiNguyen2 CATTCAAATG AAGGCCGCAG CATTGAAATC AGCCCAACCT CGACTTTTCG ThaiNguyen1 GGTTGGACGG TGGCATCAGT ACACAAGAGG AGAACACAGA GAGGCACACC ThaiNguyen2 GGTTGGACGG TGGCATCAGT ACACAAGAGG AGAACACAGA GAGGCACACC ThaiNguyen1 ACCGAGGATG TCTCTCCAAG TATGCATACT CTACTTGGAG TCAAGAACAT ThaiNguyen2 ACCGAGGATG TCTCTCCAAG TATGCATACT CTACTTGGAG TCAAGAACAT. ThaiNguyen1 G ThaiNguyen2 G H nh 3. Tr nh tù nucleotit vïng m hãa cña gien CP virót PVY ë hai méu khoai t y nhiôm bönh VÞ trý sai kh c trong tr nh tù nucleotit vïng m hãa gien CP cña 2 méu nghiªn cøu STT VÞ trý ThaiNguyen1 ThaiNguyen T C 2 26 A G 3 39 A G 4 50 C A 5 70 T C 6 86 G A 7 99 T C T A B ng 1 Hai méu khoai t y nghiªn cøu cã sù sai kh c vò tr nh tù gien CP cña virót PVY vïng m hãa ë 8 vþ trý 15, 26, 39, 50, 70, 86, 99, 111 (h nh 3, b ng 1). Do vëy, tr nh tù gien cña 2 méu nµy cã é t ng ång cao 99%. Chóng t«i tiõn hµnh so s nh hai tr nh tù axit amin cña protein CP cña virót PVY ph n lëp tõ Th i Nguyªn víi mét sè tr nh tù axit amin cña protein CP cña virót PVY îc c«ng bè trªn NCBI cã m sè AM236801, AY841269, DQ119141, AM236793, AM411504, AY742725, ThaiNguyen1, ThaiNguyen2, AY841268, AM411502, AM411503, AY [4-8]. KÕt qu cho thêy, hai méu nghiªn cøu cã é t ng ång cao (tõ 92,1-98,9%) so víi 10 méu trªn Ng n hµng gien NCBI. MÉu ThaiNguyen1 t ng ång cao nhêt víi méu cã m sè AY (97,8%) vµ thêp nhêt víi m sè AY (92,5%). KÕt qu îc thó hiön ë b ng 2. 84 B ng 2 HÖ sè t ng ång vò tr nh tù axit amin cña protein CP ë hai méu nghiªn cøu víi 10 méu trªn Ng n hµng gien NCBI HÖ sè kh c biöt HÖ sè t ng ång ,1 95,5 97,0 94,4 94,0 95,9 96,6 93,6 96,3 97,0 96,4 1 ThaiNguyen2 2 1,9 96,6 97,4 93,3 92,9 97,0 97,8 92,5 95,5 97,4 97,6 2 ThaiNguyen1 3 4,6 3,5 97,0 93,3 92,9 98,9 98,5 92,5 95,5 97,8 97,6 3 AM pro 4 3,1 2,7 3,1 94,4 94,0 97,4 98,1 92,9 95,5 98,5 97,6 4 AM pro 5 5,8 7,1 7,1 5,8 98,9 93,6 94,8 97,0 96,6 94,0 94,1 5 AM pro 6 6,3 7,5 7,5 6,3 1,1 93,3 94,4 97,4 96,3 93,6 93,7 6 AM pro 7 4,2 3,1 1,1 2,7 6,7 7,1 98,9 92,1 95,9 98,1 97,6 7 AM pro 8 3,5 2,3 1,5 1,9 5,4 5,8 1,1 93,3 97,0 98,5 98,0 8 AY pro 9 6,7 7,9 7,9 7,5 3,1 2,7 8,3 7,1 95,1 93,3 92,9 9 AY pro 10 3,8 4,6 4,6 4,6 3,5 3,8 4,2 3,1 5,0 95,9 95,7 10 AY pro 11 3,1 2,7 2,3 1,5 6,3 6,7 1,9 1,5 7,1 4,2 98,4 11 AY pro 12 3,6 2,4 2,4 2,4 6,2 6,6 2,4 2,0 7,5 4,5 1,6 12 DQ pro Tõ kõt qu so s nh tr nh tù axit amin cña protein CP ë virót PVY ë trªn, chóng t«i thó hiön kõt qu trªn bióu å h nh c y (h nh 4). H nh 4. BiÓu å h nh c y so s nh møc t ng ång cña protein CP ë 2 méu nghiªn cøu vµ 10 méu trªn Ng n hµng gien NCBI H nh 4 cho thêy, 12 méu so s nh chia thµnh 2 nhãm: nhãm I gåm 9 méu (AM236801, AY841269, DQ119141, AM236793, AM411504, AY742725, ThaiNguyen1, ThaiNguyen2, AY841268), nhãm II gåm 3 méu (AM411502, AM411503, AY742732). Trong nhãm I, méu ThaiNguyen1 vµ ThaiNguyen2 øng trong cïng nhau mét nhãm nhá vµ t ng ång cao (98,1%). 3. So s nh tr nh tù axit amin cña protein CP ë hai méu nghiªn cøu Do tr nh tù nucleotit vïng m hãa cña gien CP ë 2 méu nghiªn cøu cã é t ng ång cao nªn tr nh tù axit amin cña protein CP ë hai méu nµy còng cã é t ng ång rêt cao. KÕt qu îc thó hiön ë h nh ThaiNguyen1 ANDTIDAGES NKKDAKPEQG SIQSNLNKGK DKDVNAGTSG THTVPRIKAI ThaiNguyen2 ANDTIDAGGS NKKDAKQEQG SIQPNLNKEK DKDVNAGTSG THTVPRIKAI ThaiNguyen1 TSKMRMPKSK GATVLNLEHL LEYAPQQIDI SNTRATQSQF DAWYEAVRMA ThaiNguyen2 TSKMRMPKSK GATVLNLEHL LEYAPQQIDI SNTRATQSQF DAWYEAVRMA ThaiNguyen1 YDIGETEMPT VMNGLMVWCI ENGTSPNVNG VWVMMDENEQ VEYPLKPIVE ThaiNguyen2 YDIGETEMPT VMNGLMVWCI ENGTSPNVNG VWVMMDENEQ VEYPLKPIVE 85 ThaiNguyen1 NAKPTLRQIM AHFSDVAEAY IEMRNKKEPY MPRYGLIRNL RDVGLARYAF ThaiNguyen2 NAKPTLRQIM AHFSDVAEAY IEMRNKKEPY MPRYGLIRNL RDVGLARYAF ThaiNguyen1 DFYEVTSRTP VRAREAHIQM KAAALKSAQP RLFGLDGGIS TQEENTERHT ThaiNguyen2 DFYEVTSRTP VRAREAHIQM KAAALKSAQP RLFGLDGGIS TQEENTERHT ThaiNguyen1 TEDVSPSMHT LLGVKNM ThaiNguyen2 TEDVSPSMHT LLGVKNM H nh 5. So s nh tr nh tù axit amin cña protein CP ë hai méu nghiªn cøu B ng 3 VÞ trý sai kh c trong tr nh tù axit amin cña protein CP ë hai méu nghiªn cøu STT VÞ trý ThaiNguyen1 ThaiNguyen2 1 9 E G 2 18 P Q 3 24 S P 4 29 G E Tr nh tù axit amin cña protein CP ë 2 méu nghiªn cøu chø sai kh c nhau ë 4 vþ trý lµ 9, 18, 24, 29 (h nh 6, b ng 3). KÕt qu ph n lëp vµ x c Þnh tr nh tù gien CP cña virót PVY cña hai méu khoai t y Th i Nguyªn ë trªn lµ c së Ó chóng t«i cã thó thiõt kõ vðct chuyón gien kh ng virót g y bönh trªn khoai t y trång t¹i Th i Nguyªn. III. KÕT LUËN Chóng t«i nh n îc gien CP b»ng ph n øng PCR víi cæp måi Æc hiöu. S n phèm PCR îc dßng ho nhê vðct pbt. o¹n m hãa dµi 801 nucleotit vµ m hãa s n phèm protein dµi 267 axit amin. So s nh tr nh tù axit amin cña protein CP ë 2 méu nghiªn cøu víi 10 méu trªn Ng n hµng gien quèc tõ NCBI cho thêy é t ng ång cao (trªn 92%). TµI LIÖU THAM KH O 1. êng Hång DËt, 2005: C y khoai t y vµ kü thuët th m canh t ng n ng suêt, Nxb. Lao éng - X héi. 2. Tr ng V n Hé vµ cs., 1990: iòu tra vò b o qu n khoai t y gièng ë ång B»ng B¾c Bé, Mét sè kõt qu nghiªn cøu khoa häc c y khoai t y ( ): Nxb. N«ng nghiöp, Hµ Néi. 3. Misisiou A. et al., 2004: Molecular breeding, 14: Nguyen H. M. et al., 2006: Accession AM411502, Potato virus Y partial gene for polyprotein, genomic RNA, isolate VNY- Thaibinh. ncbi.nlm.nih.gov. 5. Nguyen H. M., Chu H. H., Pham V. T., Le B. T., 2006: Accession AM411503, genomic RNA, isolate VNY-Namdinh. 6. Nguyen H. M., Chu H. H., Pham V. T., Le B. T., 2006: Accession AM411504, genomic RNA, isolate VNY-Hanoi. ncbi.nlm.nih.gov. 7. Tian Y., Liu J., Zhu X., Li X., Valkonen J. P. T., 2008: Accession AM236793, coat protein region,genomic RNA, isolate Anhui2. ncbi.nlm.nih.gov. 8. Tian Y., Liu J., Zhu X., Li X., Valkonen J. P. T., 2008: Accession AM236801, coat protein region,genomic RNA, isolate Gansu5. ncbi.nlm.nih.gov. 86 CLONING AND SEQUENCING OF THE GENES ENCODING COAT PROTEIN (CP) IN POTATO VIRUS Y FROM THAI NGUYEN province NGUYEN THI TAM, CHU HOANG MAU, NGUYEN VU THANH THANH SUMMARY Potato (Solanum tuberosum L.) is the most widely distributed crop in tropical and subtropical zones of the world. Among diseases potato viruses play major role in reducing the yield. PVX (Potato virus X), PVY (Potato virus Y), PLRV (Potato leaf roll virus), PVA (Potato virus A), PVM (Potato virus M), PVS (Potato virus S) are major viral diseases occurring in Vietnam. PVY (Potato virus Y) is the type member of the genus Potyvirus of the family Potyviridae. The virus has a long filamentous particle containing one single-stranded, positive sense RNA genome of approximately 9.7 kb. PVY is an important pathogen infecting potato, tobacco, tomato and other solanaceous plant species. Different potato PVY isolates occur that can be categorized in 5 groups: PVY O, PVY N, PVY NTN, PVY N Wi and PVY C. In this paper we report cloning and sequencing of coat protein (CP) gene of PVY from Thai Nguyen potato. The CP gene of this isolate was found to be 801 nucleotides long, coding for 267 amino acids. Comparison amino acid of CP protein in potato from Thai Nguyen with ten sequences on NCBI showed that amino acid was % homology. Ngµy nhën bµi:
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