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棉铃虫核型多角体病毒Ha105的序列分析及敲除和回复菌株的构建

Sequence Analysis of Ha105 and Struction of Ha105's Deleted and Retrieved Recombinant Virus
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摘要 通过生物信息学方法对Ha105的生物功能进行预测,运用λ噬菌体Red重组系统介导的同源重组,在大肠杆菌BW25113中,用含有350 bp同源臂的氯霉素抗性基因和绿色荧光蛋白基因替换了棉铃虫病毒细菌人工染色体HaBacHZ8上的Ha105基因,然后利用Bac-to-Bac系统把Ha105回复到Ha105缺失的重组病毒上,构建了Ha105的缺失和回复重组病毒。生物信息学分析结果表明,Ha105是bro基因家族成员,含有Bro-N结构域,可能对宿主细胞的转录和病毒复制有一定影响。此外,重组病毒的PCR及酶切结果表明,成功构建了vHaHa105-KO-PH-gfp缺失菌株和vHaHa105-REP-PH-gfp、vHaHa105-REP-gfp回复菌株。该Ha105缺失及回复菌株的获得为进一步研究Ha105的功能奠定基础。 This paper have predicted the biological function of orf105 of Helicoverpa armigera singly-enveloped nucleopolyhedrovirus（HaSNPV）by bioinformatics.The Ha105 gene of HaSNPV bacterial artificial chromosome（HaBacHZ8） was replaced with a DNA fragment containing a chloramphenicol antibiotic gene,gfp genes and a 600 bp flanking region using the λ phage Red recombination system in E.coli BW25113.Ha105 gene were then translocated to the Ha105 deleted HaBacHZ8 using a Bac-to-Bac system.Bioinformatics analysis indicated that Ha105 belong to bro gene,including Bro-N domain.It may influence transcription of host cell and virus replication.In addition,the results of PCR and restriction show the recombinant virus have been constructed successfully.The deleted and retrieved recombinant virus made preparations for further study.
作者 曾利平 闫尧 徐旭士
机构地区 江苏省生物多样性与生物技术重点实验室南京师范大学生命科学学院
出处 《生物技术通报》 CAS CSCD 北大核心 2011年第7期134-138,共5页 Biotechnology Bulletin
关键词 棉铃虫核型多角体病毒 Bro基因 基因敲除 Red-ET重组 Bac-to-Bac昆虫表达系统 HaSNPV Bro gene Gene knockout Red-ET recombination system Bac-to-Bac system
分类号 S476.1 [农业科学—农业昆虫与害虫防治]
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参考文献8

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生物技术通报
2011年 第7期

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