痘苗病毒基因在表达四环素阻遏物的哺乳动物细胞系中的条件性表达。

Conditional expression of vaccinia virus genes in mammalian cell lines expressing the tetracycline repressor.

作者信息

Hedengren-Olcott Marika, Hruby Dennis E

机构信息

Department of Microbiology, Oregon State University, 220 Nash Hall, Corvallis, OR 97331, USA.

出版信息

J Virol Methods. 2004 Sep 1;120(1):9-12. doi: 10.1016/j.jviromet.2004.03.016.

DOI:10.1016/j.jviromet.2004.03.016

PMID:15234804

Abstract

A new system was developed for producing conditional-lethal vaccinia virus mutants using the tetracycline controlled gene expression system from bacteria. The tetracycline resistance operon (tetO) sequence was placed between the promoter and coding sequence of the target gene of the virus. To regulate the expression of the target gene, the tetO-containing virus was used to infect a tetracycline repressor (TetR) expressing cell line. This method allowed isolation of a tetO-containing virus in the absence of the TetR thereby eliminating the risk of selecting for an inactivated viral tetR gene caused by recombination of the virus genome and improving the stability of the engineered mutants.

摘要

利用细菌的四环素调控基因表达系统，开发了一种用于生产条件致死痘苗病毒突变体的新系统。四环素抗性操纵子（tetO）序列被置于病毒靶基因的启动子和编码序列之间。为了调控靶基因的表达，将含有tetO的病毒用于感染表达四环素阻遏物（TetR）的细胞系。该方法允许在不存在TetR的情况下分离含有tetO的病毒，从而消除了因病毒基因组重组导致选择失活病毒tetR基因的风险，并提高了工程突变体的稳定性。