Synthesis of encephalomyocarditis virus in a cell-free system: from DNA to RNA virus in one tube

被引：0
作者：
Tominari Kobayashi
Yusuke Nakamura
Satoshi Mikami
Mamiko Masutani
Kodai Machida
Hiroaki Imataka
机构：
[1] University of Hyogo,Department of Materials Science and Chemistry, Graduate School of Engineering
[2] RIKEN Systems and Structural Biology Center,Molecular Nanotechnology Research Center, Graduate School of Engineering
[3] University of Hyogo,undefined
来源：
Biotechnology Letters | 2012年 / 34卷
关键词：
Cell-free; DNA; Encephalomyocarditis virus; RNA virus; Virus particles; Virus synthesis;
D O I：
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中图分类号：
学科分类号：
摘要：
Virus particles are used in vaccination, drug delivery, and material sciences. Here we devised a system where the RNA virus encephalomyocarditis virus (EMCV) is synthesized from DNA templates in vitro. When a plasmid or a PCR product harboring the full-length cDNA of EMCV in the T7 promoter/terminator unit was incubated in a HeLa cell extract supplemented with T7 RNA polymerase, EMCV was produced within 4 h at an efficiency of over 10-fold compared with the system programmed with the EMCV RNA. The EMCV RNA transcribed by the virally encoded RNA-dependent RNA polymerase was predominantly incorporated into the EMCV particle even in the presence of a larger amount of the EMCV RNA transcribed by T7 RNA polymerase from the plasmid.
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页码：67 / 73
页数：6
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