H_5N_1亚型禽流感病毒核蛋白基因的克隆及分子进化分析  

Cloning and phylogenetic analysis of nucleoprotein gene of an AIV isolate (H_5N_1)

作　　者：王振国[1] 金宁一[1] 马鸣潇[1] 金扩世[1] 张洪勇[1] 尹革芬[1] 葛淑敏[1] 

机构地区：[1]解放军军需大学研究所解放军基因工程重点实验室,吉林长春130062

出　　处：《中国预防兽医学报》2004年第6期436-438,共3页Chinese Journal of Preventive Veterinary Medicine

摘　　要：用RT＿PCR法,以1株H5N1亚型禽流感病毒RNA为模板,扩增了NP全基因cDNA。将cDNA克隆于pMD18＿T载体,并对其进行测序。测序结果表明所克隆的1542个核苷酸的片段包含了核蛋白完整阅读框架,其中编码区为1497个核苷酸,编码498个氨基酸。将其序列与6株A型流感病毒NP基因序列进行比较,各毒株之间NP基因核苷酸序列同源性为92 5%～95 9%,编码的氨基酸同源性为97 0%～98 4%。本研究通过分子进化分析揭示了各毒株间的亲源关系。Complete nucleoprotein gene was amplified from an avian influenza virus(H_5N_1) by RT_PCR.The cDNA of NPgene was cloned into pMD18_T vector and then was sequenced.The cloned 1?542?bp fragment covered open reading frame of NP gene and encoded 498 amino acids.The results of comparative sequence analysis indicated that the nucleotide sequence of NP gene of this isolate shared 92.7?%～97.3?% homology with other six type A influenza virus isolates(H_5N_1) and putative amino acid sequence shared 97.0?%～98.4?% homology with other six isolates.The phylogenetic analysis revealed the relationships among these isolates.

关 键 词：RT-PCR 禽流感病毒 NP基因 分子进化分析 

分 类 号：S852.65[农业科学—基础兽医学] Q785[农业科学—兽医学]