【摘要】 本文报道应用聚合酶链式反应(PCR)技术,在体外扩增大豆花叶病毒外壳蛋白基因及其克隆和序列分析的结果。大豆花叶病毒的 RNA 从纯化的病毒粒子中提取,经逆转录合成了cDNA 的第一条链。以此为模板,利用 PCR 技术,经30个循环的扩增,得到一个特异的0.8kb片段。克隆后,对此片段进行了限制性内切酶物理图谱的分析,并测定了其全序列。实验结果证明,作者克隆到的是一个完整的大豆花叶病毒外壳蛋白基因,其核苷酸序列与已发表的大豆花叶病毒 N 株的同源率为96%,编码氨基酸的同源率为96.3%。还对利用大豆花叶病毒外壳蛋白基因培育抗大豆花叶病毒的转基因大豆的可行性进行了讨论。更多还原

【Abstract】 The results of cloning and sequencing of the gene encoding coat protein (CP) of soybean mosaic virus with PCR technique are reported. The virus RNA was extracted from purified viral particles. The first strand of cDNA was synthesized from viral RNA template primed with syn- thetic 3’polymerase chain reaction (PCR) primer using AMV reverse ,ranscriptase. A DNA fragment with O.s kb was obtained after 30 PCR amplification cycles, The restriction map of the DNA fragment has been analyzed and i~+s whole DNA sequence has been determined. The results show that the entire gene encoding the coat protein of soybean mosaic virus (SMV) has been cloned. The homologies of the DNA sequences and the deduced amino acid sequence be- tween the SMV Sa strain shown here and the N strain of SMV published abroad are 96% and 96.6% respectively. The work of tranferring the CP gene into soybean to obtain transgenic soy- bean plants that resist to SMV infection is in progress.更多还原