A Novel Bacterium-Free Method for Generation of Flavivirus Infectious DNA by Circular Polymerase Extension Reaction Allows Accurate Recapitulation of Viral Heterogeneity

被引：63
作者：
Edmonds, Judith [1 ]
van Grinsven, Erinke [1 ,2 ]
Prow, Natalie [1 ]
Bosco-Lauth, Angela [3 ]
Brault, Aaron C. [3 ]
Bowen, Richard A. [4 ]
Hall, Roy A. [1 ]
Khromykh, Alexander A. [1 ]
机构：
[1] Univ Queensland, Sch Chem & Mol Biosci, Australian Infect Dis Res Ctr, Brisbane, Qld, Australia
[2] Univ Utrecht, Grad Sch Life Sci, Utrecht, Netherlands
[3] Ctr Dis Control & Prevent, Div Vector Borne Dis, Ft Collins, CO USA
[4] Colorado State Univ, Ft Collins, CO 80523 USA
来源：
JOURNAL OF VIROLOGY | 2013年 / 87卷 / 04期
基金：
英国医学研究理事会;
关键词：
WEST-NILE-VIRUS; JAPANESE-ENCEPHALITIS-VIRUS; FULL-LENGTH CDNA; ENVELOPE PROTEIN; KUNJIN VIRUS; IN-VITRO; RT-PCR; RNA; STRAIN; CLONE;
D O I：
10.1128/JVI.03162-12
中图分类号：
Q93 [微生物学];
学科分类号：
071005 ; 100705 ;
摘要：
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.
引用
收藏
页码：2367 / 2372
页数：6
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