一种新颖的无细菌环聚合酶延伸反应生成黄病毒感染性 DNA 的方法可准确再现病毒异质性。
A novel bacterium-free method for generation of flavivirus infectious DNA by circular polymerase extension reaction allows accurate recapitulation of viral heterogeneity.
机构信息
Australian Infectious Disease Research Centre, School of Chemistry and Molecular Biosciences, University of Queensland, Brisbane, Queensland, Australia.
出版信息
J Virol. 2013 Feb;87(4):2367-72. doi: 10.1128/JVI.03162-12. Epub 2012 Dec 12.
Abstract
A novel bacterium-free approach for rapid assembly of flavivirus infectious cDNAs using circular polymerase extension reaction was applied to generate infectious cDNA for the virulent New South Wales isolate of the Kunjin strain of West Nile virus (KUNV) that recently emerged in Australia. Recovered virus recapitulated the genetic heterogeneity present in the original isolate. The approach was utilized to generate viral mutants with designed phenotypic properties and to identify E protein glycosylation as one of the virulence determinants.
摘要
一种新颖的无细菌方法，利用环聚合酶延伸反应快速组装黄病毒感染性 cDNA，应用于产生最近在澳大利亚出现的具有强毒力的新南威尔士州 Kunjin 株西尼罗河病毒（KUNV）的感染性 cDNA。回收的病毒重现了原始分离株存在的遗传异质性。该方法用于产生具有设计表型特性的病毒突变体，并确定 E 蛋白糖基化是一个毒力决定因素。
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