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致倦蜚蠊浓核病毒(PfDNV)作为一种用于体内持续性外源基因表达的昆虫载体。

The densovirus of Periplaneta fuliginosa (PfDNV) as an insect vector for persistent foreign gene expression in vivo.

作者信息

Hu Liu, Zhang Lei, Shen Chao, Lu Jie, Zhang Jiamin, Hu Yuanyang

机构信息

State Key Laboratory of Virology, College of Life Sciences, Wuhan University, Wuhan 430072, China.

出版信息

Biochem Biophys Res Commun. 2007 Jul 13;358(4):976-82. doi: 10.1016/j.bbrc.2007.04.210. Epub 2007 May 11.

Abstract

An infectious clone of the Periplaneta fuliginosa densovirus (PfDNV) has been constructed and the PfDNV genome can rescue from the plasmid and replicate as the wild-type virus in nymphs of P. fuliginosa. To investigate the ability of the cloned PfDNV genome to be used as a stable and persistent expression vector, we constructed seven recombinant plasmids in which the GFP reporter gene was inserted into the genome of PfDNV. When these recombinant constructs were transfected into hosts, the GFP was expressed efficiently in every clone. Southern blot analysis revealed that recombinant plasmids had integrated into host genome. Infectious recombinant virions could be produced from plasmids in which the GFP gene was downstream of and in frame with the NS3 and NS1 coding regions. These results indicate that PfDNV genome can be used as an insect vector for the transfer and persistent expression of an exogenous gene.

摘要

已构建了黑胸大蠊浓核病毒(PfDNV)的感染性克隆,PfDNV基因组可从质粒中拯救出来,并在黑胸大蠊若虫中作为野生型病毒进行复制。为了研究克隆的PfDNV基因组作为稳定和持久表达载体的能力,我们构建了七个重组质粒,其中绿色荧光蛋白(GFP)报告基因被插入到PfDNV基因组中。当这些重组构建体转染到宿主中时,每个克隆中GFP都能高效表达。Southern印迹分析表明重组质粒已整合到宿主基因组中。在GFP基因位于NS3和NS1编码区下游且读码框一致的质粒中可产生感染性重组病毒粒子。这些结果表明PfDNV基因组可作为昆虫载体用于外源基因的转移和持久表达。

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