Based on multi-alignment of complete polyprotein amino acid sequences of genus Potyvirus,five degenerated primers were designedThey were Sprimer(5′-GGX AAY AAY AGY GGX CAZ CC-3′),pNIa(+)(5′-TNY TGG AAM CAY TGG A...Based on multi-alignment of complete polyprotein amino acid sequences of genus Potyvirus,five degenerated primers were designedThey 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展开更多
通过使用blastx、Blockmaker、CODEHOP和Primer Premier 5.0等在线网络工具和生物软件,针对中国卤虫Orthodenticle基因高度保守区域设计了简并引物.用所设计的简并引物克隆了中国卤虫Orthodenticle基因片段.在GenBank中以blastx方法进...通过使用blastx、Blockmaker、CODEHOP和Primer Premier 5.0等在线网络工具和生物软件,针对中国卤虫Orthodenticle基因高度保守区域设计了简并引物.用所设计的简并引物克隆了中国卤虫Orthodenticle基因片段.在GenBank中以blastx方法进行比对,发现此段基因片段与美国卤虫Orthodenticle基因有高度的相似性.通过实验进一步证明,此种设计简并引物的方法可信性强,特异性高,能够快速得到满意的实验结果.展开更多
文摘Based on multi-alignment of complete polyprotein amino acid sequences of genus Potyvirus,five degenerated primers were designedThey 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
文摘通过使用blastx、Blockmaker、CODEHOP和Primer Premier 5.0等在线网络工具和生物软件,针对中国卤虫Orthodenticle基因高度保守区域设计了简并引物.用所设计的简并引物克隆了中国卤虫Orthodenticle基因片段.在GenBank中以blastx方法进行比对,发现此段基因片段与美国卤虫Orthodenticle基因有高度的相似性.通过实验进一步证明,此种设计简并引物的方法可信性强,特异性高,能够快速得到满意的实验结果.
