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本研究从带有黄化症状的南瓜叶片中提取总RNA,用RT-PCR方法扩增得到来自湖北和云南的南瓜蚜传黄化病毒(CABYV)2个分离物的1375nt特异性核苷酸片段。分别将PCR产物插入到克隆载体pMD19-T并转化大肠杆菌DH5α,对筛选到的阳性克隆进行了序列测定和分析(GenBank登录号为EF488996和EF488997)。所获片段含有部分复制酶基因576nt,非编码区199nt和完整的CP基因600nt,编码一个由199个氨基酸组成的分子量约为22kDa的结构蛋白。湖北和云南分离物与法国分离物、意大利分离物、西班牙分离物、北京分离物和上海分离物的CP基因核苷酸序列和推测氨基酸序列的同源性分别为93.1%~98.5%和91.4%~98.5%。
In this study, we extracted total RNA from pumpkin leaves with yellowing symptoms and amplified 1375nt specific nucleotide fragments from 2 isolates of squash aphid (CABYV) from Hubei and Yunnan by RT-PCR . The PCR products were inserted into the cloning vector pMD19-T and transformed into E. coli DH5α. The positive clones were sequenced and analyzed (GenBank accession numbers EF488996 and EF488997). The resulting fragment contained 576 nt of partial replicase gene, 199 nt of non-coding region and 600 nt of the complete CP gene, encoding a structural protein of about 22 kDa consisting of 199 amino acids. The homologies of nucleotide and deduced amino acid sequences of CP gene in Hubei and Yunnan isolates were 93.1% -98.5% and 91.4% respectively with those of French isolates, Italian isolates, Spanish isolates, Beijing isolates and Shanghai isolates. ~ 98.5%.