一株诺如病毒的3’RACE扩增及基因序列分析  

3'RACE Amplification and Sequence Analysis of a Strain of Norovirus

作　　者：李雅静[1] 林祥梅[2] 吴绍强[2] 刘建[2] 梅琳[2] 

机构地区：[1]河北师范大学生命科学学院,石家庄050016 [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：《中国畜牧兽医》2008年第8期36-40,共5页China Animal Husbandry & Veterinary Medicine

基　　金：国家科技支撑计划(2006BAD06A14);国家质检总局科研基金(2006IK038)

摘　　要：诺如病毒是严重危害人类健康的重要食源性病原。作者应用3’RACE(rapid amplification of cDNA 3’ends)技术对一株诺如病毒分离株的基因组3’末端进行扩增,并对其核苷酸序列进行比较分析,从而可为该病的分子生物学检测提供阳性物质,并从分子水平上推测该病的来源。经过提取病毒总RNA、以3’RACE锚定引物进行反转录、用特异引物进行3’RACE,琼脂糖凝胶电泳检测结果表明成功扩增了诺如病毒基因组约3282 bp的基因片段,测序及序列BLAST分析证实该分离株属GⅡ-4型,与日本分离株同源。3’RACE扩增序列为诺如病毒的实验室检测及深入研究奠定了基础。Norovirus is an important zoonotic pathogen threatening human health.To serve as positive control for the molecular biological examination of norovirus infection and facilitate deducing the origin of this isolate,the total RNA of a feces sample was extracted and used as template for reverse transcription.The first strand cDNA was amplified by 3 RACE（rapid amplification of cDNA 3 ends） using gene specific primer JV12 as forward inner primer and 3 RACE backward inner primer.The amplicon was 3282 bp in length judged from agarose electrophoresis. The amplicon was ligated to the Promega T-easy vector and sequence determined. The determined nucleotide sequence was then analyzed by blast online. Sequence analysis indicated that the isolated virus strain belonged to Norovirus GⅡ-4 cluster and has the same origin with the isolate in Japan. 3 ＇RACE amplification established a basis for experimental detection and further research of Norovirus.

关 键 词：诺如病毒 3’RACE 序列分析 起源 

分 类 号：Q785[生物学—分子生物学]