葡萄卷叶伴随病毒基因的克隆及序列分析

Gene clone and nucleotid sequence analysis of grapevine leafroll virus 3

根据葡萄卷叶伴随病毒 美国分离物的 GLRa V- 3CP基因序列 ,设计并合成了 1对该病毒的 PCR引物 ,利用 IC- RT- PCR成功地检查到合适大小的 PCR产物 ;同时将PCR产物克隆到中间载体 PGEM- 3Zf上 ,转化大肠杆菌 DH5α菌株 ,筛选阳性克隆。通过双酶切以及序列分析表明 ,扩增样品与美国分离物的同源性为 94.1 %。

A pair of primers were designed and synthesized based on the nucleotide sequence of coat protein of GLRav 3 from America.By IC RT PCR method,the expected size PCR products were amplified.This product was cloned into vector PGEM3Zf,then translated into E.Coli BL2l,and the recombinant plasmid has been obtained.The nucleotid sequence analysis of PCR products showed 94.1% of the nucleotid sequence is identical with nucleotid sequence of coat protein of GLRaV 3 from America.