李属坏死环斑病毒怀柔分离物(PNRSV-HR)外壳蛋白基因片断的克隆与序列分析  

The cloning and sequence analysing of the CP gene from huairou isolate of Prunus necrotic ringspot virus

作　　者：黄冲[1] 李明福[2] 张永江[2] 

机构地区：[1]中国农业大学植物病理学系,北京100094 [2]中国检验检疫科学研究院动植物检疫研究所,北京100029

出　　处：《西北农林科技大学学报（自然科学版）》2005年第B08期105-108,共4页Journal of Northwest A&F University(Natural Science Edition)

基　　金：国家自然科学基金项目(30270892)

摘　　要：应用RT-PCR方法检测了采自北京市怀柔区的樱桃叶片,将此PCR扩增产物连接到pMD18-T载体上,通过转化大肠杆菌DH5α感受态细胞,克隆了含有目的片段的重组质粒,并进行序列测定和分析。序列分析结果表明,认为该分离物外壳蛋白基因片断与李属坏死环斑病毒的NRSiz8株系的同源性最高,为98%,与德国分离物Ring17的同源性达99%;通过进化关系分析,认为该分离物属于引起温和症状的group,认为该分离物的第5,62,81和126位氨基酸也与CH9血清型引起温和症状的保守的替换相同。The leaves of cherry were sampled from Huairou,Beiiing and detected by RT-PCR amplification. The PCR product was cloned in pMD18-T vector,and transformed to the DH5α. The recombinant plasmid was analysed with Sanger method. The result showed that the CP-gene sequence had basic harmonization with that of strain NRSiz8 （98% homology） and isolate Ring 17 （99% homology）. The phylogenetic trees suggest that PNRSV-HR belongs to group 11 （the mild pathotype,defined by Hammond） of PNRSV,and the 5th ,62th,81th ,and 126th amino acid of the isolate are same to the CH9 serotype.

关 键 词：李属坏死环斑病毒(PNRSV) RT—PCR 克隆 序列分析 

分 类 号：S436.629[农业科学—农业昆虫与害虫防治]