文档介绍：含SARS CoV RNA病毒样颗粒的构建和表达
作者:李振勇,景建洲,刘明霞,邵大晓,冯艳铭,李智涛,宋国英,王秋旗,王云龙,屈凌波,赵玉芬
【关键词】病毒样颗粒
【Abstract】 AIM: To construct and express RNaseresistant viruslike particles containing the RNA fragments of SARS CoV. METHODS: The primers of MS2 assembly protein gene, coat protein gene and the fragment of SARS Cov RNA polymerase gene were synthesized according to their sequences from Genbank data and were used to amplify their cDNAs by RTPCR. These cDNAs were purified with SABCs ResincolumTM system for DNA purification from agarose gel. Expression vector pTrc99a and their cDNA fragments were ligated together with T4 DNA ligase after digestion with restriction enzymes. The prokaryotic expression was obtained by the transformation of the binant plasmid constructed above into JM109. The obtained viruslike particles were purified and tested by quantitative analysis, RTPCR and stable experiment. RESULTS: The RNaseresistant viruslike particles containing the RNA fragments of SARS CoV was essfully constructed and remained stable for 30 days at 37
℃. CONCLUSION: The newtype viruslike particles can be conveniently used as positive control of SARS clinical nucleic diagnostic reagents.
【Keywords】 SARS; reverse transcriptase polymerase chain reaction; RNA, viruslike particles; ribonucleases
【摘要】目的: 构建并表达含有SARS冠状病毒RNA片断的抗核糖核酸酶的病毒样颗粒. 方法: 通过克隆大肠杆菌噬菌体MS2的装配蛋白和壳蛋白基因以及SARS冠状病毒RNA聚合酶基因片段,将这些基