[1]耿宏伟,郭怡璠,万书明,等.马铃薯Y病毒黑龙江分离株外壳蛋白基因克隆及序列分析[J].黑龙江农业科学,2015,(02):11-13.[doi:10.11942/j.issn1002-2767.2015.02.00011]
　GENG Hong-wei,GUO Yi-fan,WAN Shu-ming,et al.Cloning and Sequences Analysis of Heilongjiang Isolate Potato Virus Y CP Gene[J].HEILONGJIANG AGRICULTURAL SCIENCES,2015,(02):11-13.[doi:10.11942/j.issn1002-2767.2015.02.00011]
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马铃薯Y病毒黑龙江分离株外壳蛋白基因克隆及序列分析

《黑龙江农业科学》[ISSN:1002-2767/CN:23-1204/S] 卷: 期数: 2015年02 页码: 11-13 栏目: 出版日期: 2015-02-10
Title:
Cloning and Sequences Analysis of Heilongjiang Isolate Potato Virus Y CP Gene
文章编号:
1002-2767（2015）02-0011-03
作者:
耿宏伟; 郭怡璠; 万书明; 范国权; 魏琪; 高艳玲; 张威
黑龙江省农业科学院 植物脱毒苗木研究所
Author(s):
GENG Hong-wei; GUO Yi-fan; WAN Shu-ming; FAN Guo-quan; WEI Qi; GAO Yan-ling; ZHANG Wei
Virus-free Seeding Research Institute of Heilongjiang Academy of Agricultural Sciences
关键词:
RT-PCR; 马铃薯Y病毒; 序列分析
Keywords:
RT-PCR; potato virus Y; sequence analysis
分类号:
S532
DOI:
10.11942/j.issn1002-2767.2015.02.00011
文献标志码:
A
摘要:
为研究马铃薯Y病毒的检测方法，参考GenBank中马铃薯Y病毒的全基因序列，应用Primer 6.0引物软件设计并合成了一对特异性引物PVYF、PVYR，利用RTPCR方法对PVY病毒黑龙江分离株CP基因进行特异性扩增，对扩增片段进行序列测定并进行序列比对。结果表明：引物PVYF、PVYR可以扩增出包含PVY病毒CP基因全长的cDNA特异性片段，序列分析结果表明PVY病毒黑龙江分离株CP基因与GenBank上的40个不同PVY病毒分离株的CP基因氨基酸序列同源性为92.1%~97.4%，说明PVY病毒CP基因保守性较高，可以用作制备PVY病毒血清型检测方法的抗体基因。
Abstract:
For the research of potato virus Y,pair of primers PVYF,PVYR were designed according to the complete genome of potato virus Y (PVY) in GenBank,the PVY CP gene was amplified from strain isolated from Heilongjiang.After that,the information of amino acid sequences with computer technology was studied.The RT-PCR results showed that the primers could be amplified the CP gene fragment.The analysis of sequence result showed that the CP gene amino acid homology of Heilongjiang isolate and other 40 isolates of PVY in GenBank were 92.1%~97.4%,CP gene was highly conserved that could be used as antibody gene to detect the PVY virus.

参考文献/References:

［1］高秀妍，高艳玲，耿宏伟，等.马铃薯Y病毒黑龙江分离物株系鉴定[J].中国马铃薯，2009，23(1):11-14.
[2]Salazar L F.Potato viruses and their control[J].International Potato Center,Lima,Peru,1996:616. [3]Hu Xiaojun,Alexander V K,Celeste J B,et al.Sequence characteristics of potato virus Y recombinants\[J\].Journal of General Virology,2009,90:3033-3041.
[4]AstierManifacier，Francine CasseDelbart.Nucleotide sequence of potato vrius Y (N strain)genomic RNA\[J\].Journal of General Virology,1989,70:935947.
[5]白艳菊，李学湛，何云霞，等.优质、低成本工厂化生产马铃薯脱毒试管苗[J].中国农学通报，2001，17(2):82-83.
[6]张威，张匀华，李学湛，等.应用RT-PCR分子检测技术快速检测大蒜普通潜隐病毒[J].植物保护，2008，34(1):133-137.
[7]耿宏伟，白艳菊，范国权，等.应用RT-PCR技术检测马铃薯A病毒[J].中国马铃薯，2009（6）：329331.
[8]Zhang Y,Guo D,Geng H,et al.Characterization of Mclass genome segments of Muscovy duck reovirus S14[J].Virus Research,2007,125:42-53.
[9]Rouis S,Lafaye P,JaouaAydi L,et al.Cloning and expression of functional singlechain Fv antibodies directed against NIa and coat proteins of potato virus Y[J].Journal of Virological Methods,2006,137:1-6.
[10]Nie X,Singh R P.Detection of multiple potato viruses using an oligo(dT) as a common cDNA primer inmultiplex RT-PCR[J].Journal of Virological Methods,2000,86:179-185.
[11]Gugerli P.Potato virus A and potato leafroll virus:purification,antiserum production and serological detection in potato and test plants by Enzymelinked Immunosorbent Assay (ELSIA)[J].Journal of Phytopathology,1979,96(2):97-107.
[12]Donald J G,Kristjansson G T,Singh R P,et al.Consecutive ELISA screening with monoclonal antibodies to detect potato virus YN[J].Amercan Journal of Potato research,1994,71(3):175-183.
[13]Kohler G，Milstein C.Continuous cultures of fused cells secreting antibody of predefined specificity[J ].Nature,1975,256:495-497.

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备注/Memo

基金项目：〖HTSS〗黑龙江省自然科学基金资助项目（C201120）；2010年黑龙江省农业科学院青年基金资助项目；哈尔滨市科技创新人才资助项目（2014RFQYJ050）〖ZK）〗〖ZK(〗〖HTH〗第一作者简介：〖HTSS〗耿宏伟（1982-），男，黑龙江省哈尔滨市人，博士，副研究员，从事马铃薯病害分子生物学研究。E-mail：hongwei-geng@163.com。

更新日期/Last Update: 2015-05-15