当前位置: X-MOL 学术J. Virol. Methods › 论文详情
Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)
Stabilization of a full-length infectious cDNA clone for duck Tembusu virus by insertion of an intron.
Journal of Virological Methods ( IF 3.1 ) Pub Date : 2020-06-26 , DOI: 10.1016/j.jviromet.2020.113922
Jiaqi Guo 1 , Yu He 1 , Xiaoli Wang 1 , Bowen Jiang 1 , Xiao Lin 1 , Mingshu Wang 2 , Renyong Jia 2 , Dekang Zhu 3 , Mafeng Liu 2 , Xinxin Zhao 2 , Qiao Yang 2 , Ying Wu 2 , Shun Chen 2 , Anchun Cheng 2
Affiliation  

Duck Tembusu virus (DTMUV) belongs to the genus Flavivirus, family Flaviviridae. In our previously study, a full-length cDNA clone of DTMUV was constructed, however, it is prone to mutation during genetic engineering due to the prokaryotic toxicity of viral protein, which is also a common feature for flavivirus. In this study, we reported an intron-containing full-length cDNA clone for a clinical strain CQW1, the intro (133bp) was inserted into nonstructural protein 1 of DTMUV at 192 site. This intron-containing full-length cDNA clone was stably propagated in Escherichia coli without prokaryotic toxicity, and recombinant virus was produced by direct transfection of plasmids. Besides, this cDNA clone-derived recombinant virus showed similar properties in comparison with parent virus both in vitro and in vivo. It’s convenient and efficient, making it a useful platform for the subsequent research of reverse genetics of flavivirus.



中文翻译:

通过插入内含子来稳定鸭Tembusu病毒的全长感染性cDNA克隆。

鸭坦布苏病毒(DTMUV)属于黄病毒科黄病毒科。在我们之前的研究中,构建了DTMUV的全长cDNA克隆,但是由于病毒蛋白的原核毒性,它在基因工程中易于突变,这也是黄病毒的常见特征。在这项研究中,我们报告了临床菌株CQW1的含内含子的全长cDNA克隆,该引物(133bp)已插入192位DTMUV的非结构蛋白1中。该含内含子的全长cDNA克隆在大肠杆菌中稳定繁殖没有原核毒性,并且通过直接转染质粒产生重组病毒。此外,该cDNA克隆来源的重组病毒在体外体内均与亲本病毒相比具有相似的特性。它方便,高效,使其成为黄病毒反向遗传学后续研究的有用平台。

更新日期:2020-07-06
down
wechat
bug