单引物法同时克隆RDV基因组片段S11、S12及其序列分析

Molecular Cloning Segment S11 and S12 Of RDV SynChrOnOuSIy by Single Primer Amplification and Sequence Analysis

水稻矮缩病毒(Rice dwarf wrus,RDV)6个地方分离物基因组的lO%SDS-PAGE电泳图谱显示:松溪分离物基因片段S11、S12相对迁移速率明显不同。运用单引物法同时克隆该分离物的基因组片段S11、s12,并测定它们的全序列,结果表明s11、S12全长分别是1036bp、l066bp,基因结构和报道的RDV福州分离物基本一致,但分别突变了34、24个碱基,同源性分别为97.01%、97.86%。同时单引物法也为dsRNA病毒基因组的克隆提供了一种方法。

The 10% SDS-PAGE of dsRNA profiles from six RDV isolates showed： the relative velocity of Gegment S11, S12 from RDV-SX isolate was obviously dissimilar from others. The full length cDNAs of RDV-SX isolate genome segments S11, S12 were cloned by Single Primer Amplification and their sequences were determined. The result showed the full length of S11, S12 is 1036bp, 1066bp respectively and has the same organization with RDV in Fuzhou. The similarity of nucleotide was 97.01%, 97.86% with Fuzbou isolate respectively. This study provided a good method for the sequence determination of viral dsRNA atthe sametime.