EGFP/GLUC 标记的辛德毕斯样病毒 XJ-160 的构建。

Development of EGFP/GLUC-tagged Sindbis-like virus XJ-160.

机构信息

State Key Laboratory for Infectious Disease Prevention and Control (SKLID), National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention (IVDC, China CDC), 100 Yingxin Street, Xuan Wu District, Beijing 100052, China.

出版信息

J Virol Methods. 2013 Apr;189(1):235-7. doi: 10.1016/j.jviromet.2013.01.015. Epub 2013 Feb 9.

DOI:10.1016/j.jviromet.2013.01.015

PMID:23403149

Abstract

Based on an infectious clone of Sindbis-like virus XJ-160, recombinant vectors containing a reporter gene (enhanced green fluorescence protein [EGFP] or Gaussia luciferase [GLUC]) were constructed by placing the reporter gene cassette containing the subgenomic promoter behind the 3' terminus of the viral structural protein gene. EGFP/GLUC-tagged Sindbis-like viruses were rescued in BHK-21 cells transfected with transcripts produced from the recombinant vectors. EGFP expression and strong luciferase activity were detected in BHK-21 cells infected with repeated passages of the EGFP/GLUC-tagged viruses, revealing the genetic stability of the chimeric viruses. The EGFP/GLUC-tagged Sindbis viruses reported will contribute to the assessment of viral replication and proliferation, tracking and elucidating Alphavirus-host interactions, and screening for antiviral compounds.

摘要

基于辛德毕斯样病毒 XJ-160 的感染性克隆，通过将包含亚基因组启动子的报告基因盒置于病毒结构蛋白基因的 3' 末端，构建了含有报告基因（增强型绿色荧光蛋白 [EGFP] 或高斯荧光素酶 [GLUC]）的重组载体。通过转染来自重组载体的转录本，在 BHK-21 细胞中拯救了 EGFP/GLUC 标记的辛德毕斯样病毒。在感染了重复传代的 EGFP/GLUC 标记病毒的 BHK-21 细胞中检测到 EGFP 表达和强烈的荧光素酶活性，表明嵌合病毒的遗传稳定性。报告的 EGFP/GLUC 标记辛德毕斯病毒将有助于评估病毒复制和增殖、跟踪和阐明黄病毒-宿主相互作用以及筛选抗病毒化合物。