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Position: Home > Articles > 3'RACE Amplification and Sequence Analysis of a Strain of Norovirus China Animal Husbandry & Veterinary Medicine 2008 (8) 36-37+39-40

一株诺如病毒的3’RACE扩增及基因序列分析

作  者:
LI Ya-jing;林祥梅;WU Shao-qiang;刘建;MEI Lin
单  位:
中国检验检疫科学研究院动植物检疫研究所
关键词:
诺如病毒;3’RACE;序列分析;起源
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。
译  名:
3'RACE Amplification and Sequence Analysis of a Strain of Norovirus
作  者:
LI Ya-jing1,2,LIN Xiang-mei2,WU Shao-qiang2,LIU Jian2,MEI Lin2(1.College of Life Science,Hebei Normal University,Shijiazhuang 050016,China;2.Institute of Animal and Plant Quarantine,Chinese Academy of Inspection and Quarantine,Beijing 100029,China)
关键词:
norovirus;3'RACE;sequence analysis;origin
摘  要:
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.

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