日本脑炎病毒WHe株全长cDNA的长链RT-PCR法的扩增  

Amplification of full-length cDNA of Japanese encephalitis virus WHe strain by long RT-PCR

作　　者：唐青海[1] 乔宪凤[2] 马秋明[1] 何维明[1] 郑新民[2] 刘西梅[2] 安立国[1] 颜艳[2] 魏庆信[2] 

机构地区：[1]西北农林科技大学动物科技学院,陕西杨凌712100 [2]湖北省农业科学院畜牧兽医研究所,湖北武汉430064

出　　处：《西北农林科技大学学报（自然科学版）》2007年第8期19-23,共5页Journal of Northwest A&F University(Natural Science Edition)

基　　金：湖北省自然科学基金项目(2005ABA195)

摘　　要：为深入探索日本脑炎病毒(Japanese encephalitis virus,JEV)WHe株的致病机理,并为新型疫苗研发提供技术支持,采用长链RT-PCR技术对其基因组全长cDNA进行了扩增鉴定,并对含复杂二级结构的3′末端和5′末端非编码区扩增片段进行了测序分析。扩增结果表明,扩增出的JEV WHe株基因组全长cDNA分子近11 kb大小;非编码区测序分析表明,扩增产物为WHe株所特有。提示长链RT-PCR法可用于JEV WHe株基因组全长cD-NA的扩增。In order to elucidate the pathogenesis of Japanese encephalitis virus （JEV） Wile strain and develope novel vaccine,the full-length cDNA was amplified by long reverse transcription-polymerase chain reaction（long RT-PCR）. The full-length cDNA was identified by PCR and the fragments which contain the 3rand 5runtranslation regions were cloned into pGEM-T vector, and the sequences of inserted fragments were determined by Invitrogen Corporation. The results showed that the approximate 11 kb full-length cD- NAs of WHe strain were amplified by long RT-PCR and its correctness was proved by partial nucleotide sequence analysis. The genomic cDNAs of WHe strain were successfully amplified.

关 键 词：日本脑炎病毒WHe株 全长CDNA 长链RT-PCR 

分 类 号：R373.31[医药卫生—病原生物学] S855.99[医药卫生—基础医学]