Bmp24缺失对家蚕核型多角体病毒基因组的复制、转录及病毒组装的影响  

Effect of Bmp24 Deficiency on Replication,Transcription and Virus Assembly of BmNPV

作　　者：陈琛[1] 于威[1,2] 史利利[1,2] 巩成见 蒋磊[1,2] 童富淡[1,2] 

机构地区：[1]浙江理工大学生物化学研究所,杭州310018 [2]浙江理工大学浙江省家蚕生物反应器和生物医药重点实验室,杭州310018

出　　处：《浙江理工大学学报（自然科学版）》2016年第1期109-115,共7页Journal of Zhejiang Sci-Tech University(Natural Sciences)

基　　金：国家高技术研究发展计划"863"项目(2011AA100603)

摘　　要：研究Bmp24基因的生物学功能,利用Red重组技术和Bac-to-Bac系统构建Bmp24缺失型病毒(Bmp24-ko-Bacmid)和Bmp24修复型病毒(Bmp24-re-Bacmid)。然后将Bmp24-ko-Bacmid、Bmp24-re-Bacmid以及野生型病毒(wtBacmid)分别转染BmN细胞,病毒滴度测定发现3种病毒都能产生有活力的子代病毒并使细胞发病,但Bmp24-ko-Bacmid在各个时相的滴度值显著低于其他两种病毒(P<0.05)。电镜观察结果发现Bmp24-ko-Bacmid感染的细胞中只有少量细长的杆状结构,而其他两种病毒感染细胞后能够产生大量具有囊膜包裹的成熟病毒粒子。qPCR实验结果显示,Bmp24缺失不会影响病毒基因组的复制,同时qRT-PCR结果显示Bmp24缺失使早期、晚期基因和极晚期基因的转录水平显著低于野生型病毒(P<0.05)。This research intends to study the biological function of Bmp24.We constructed Bmp24-koBacmid Red recombination technology and Bac-to-Bac system.Then,Bmp24-ko-Bacmid,Bmp24-re-Bacmid and wtBacmid transfected BmN cells.It was found through virus titer that the three viruses could generate viable progeny virus and induce cell pathogenesis,but the titerring values of Bmp24-ko-Bacmid was markedly lower than the other two types of viruses in various phases（P0.05）.Transmission electron microscopy result shows that among the cells infected by Bmp24-ko-Bacmid,there is only a small amount of long slender rod-shaped structures,while the other two virus-infected cells can produce large amounts of mature virus particles with capsule package.The qPCR result shows that lack of Bmp24 will not affect the replication of the viral genome.And the qRT-PCR result shows that lack of Bmp24 will reduce the transcription levels of early gene,late gene and very late gene which are significantly lower than that of the wild-type virus（P0.05）.

关 键 词：家蚕核型多角体病毒 Bmp24基因 RED重组系统 BAC-TO-BAC系统 

分 类 号：Q812[生物学—生物工程]