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Potyvirus属成员基因组全序列的简并引物PCR和RACE扩增方法 被引量:30

Determination of Genome Sequence of Potyviruses by Degenerated PCR and RACE Methods
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摘要 Based on multi-alignment of complete polyprotein amino acid sequences of genus Potyvirus,five degenerated primers were designedThey were Sprimer(5′-GGX AAY AAY AGY GGX CAZ CC-3′),pNIa(+)(5′-TNY TGG AAM CAY TGG AT-3′),pCI2(+)(5′-GCX ACX AAX ATX ATX GAX AA-3′),pCI1(+)(5′-GTX GGX TCX GGX AAX TCX AC-3′)and pHC(+)(5′-TGY GAY AAY CAZ TTX GA-3′)(X=A,T,C or G:Y=T or C;Z=A or G;N=A or T;M=A,T or G)Using degenerated PCR and modified RACE methods,a protocol for determination of complete genome sequence of potyviruses was established and proved to be successful on five Based on multi-alignment of complete polyprotein amino acid sequences of genus Potyvirus,five degenerated primers were designedThey were Sprimer(5′-GGX AAY AAY AGY GGX CAZ CC-3′),pNIa(+)(5′-TNY TGG AAM CAY TGG AT-3′),pCI2(+)(5′-GCX ACX AAX ATX ATX GAX AA-3′),pCI1(+)(5′-GTX GGX TCX GGX AAX TCX AC-3′)and pHC(+)(5′-TGY GAY AAY CAZ TTX GA-3′)(X=A,T,C or G:Y=T or C;Z=A or G;N=A or T;M=A,T or G)Using degenerated PCR and modified RACE methods,a protocol for determination of complete genome sequence of potyviruses was established and proved to be successful on five potyviruses
作者 陈炯 陈剑平
出处 《病毒学报》 CAS CSCD 北大核心 2002年第4期371-374,共4页 Chinese Journal of Virology
基金 浙江省青年科技人才专项(RC9604) 浙江省重点实验室专项(001107557) 浙江省十五重点科技项目(011102181)
关键词 Potyvirus属成员 基因组分序列 PCR RACE 扩增方法 马铃薯Y病毒属 简并引物 全序列测定 扩增方法 genus Potyvirus degenerated primer determination of genome sequence
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参考文献5

  • 1Jiong Chen,Jianping Chen,M. J. Adams. Characterisation of potyviruses from sugarcane and maize in China[J] 2002,Archives of Virology(6):1237~1246
  • 2J. Chen,H. Y. Zheng,J. P. Chen,M. J. Adams. Characterisation of a potyvirus and a potexvirus from Chinese scallion[J] 2002,Archives of Virology(4):683~693
  • 3H. M. Zhang,J. P. Chen,M. J. Adams. Molecular characterisation of segments 1 to 6 of Rice black-streaked dwarf virus from China provides the complete genome[J] 2001,Archives of Virology(12):2331~2339
  • 4Jiong Chen,Jianping Chen,Jishuang Chen,M. J. Adams. Molecular characterisation of an isolate of Dasheen mosaic virus from Zantedeschia aethiopica in China and comparisons in the genus Potyvirus[J] 2001,Archives of Virology(9):1821~1829
  • 5J. Chen,M. J. Adams. A universal PCR primer to detect members of the Potyviridae and its use to examine the taxonomic status of several members of the family[J] 2001,Archives of Virology(4):757~766

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