Expression of Prokaryotic Gene in Mammalian Cells Based on Recombinant Vaccinia Virus Encoding T7 RNA Polymerase

China Biotechnology

2011, Vol. 31

Issue (5): 48-54 DOI:

Expression of Prokaryotic Gene in Mammalian Cells Based on Recombinant Vaccinia Virus Encoding T7 RNA Polymerase

PAN Xiao-xia, ZHANG Shun, WEN Yu-ling, YUAN Jing, CHEN Yuan-ding

Institute of Medical Biology, Chinese Academy of Medical Sciences & Peking Union Medical College, Kunming 650118, China

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Abstract

The vaccinia virus/T7 RNA polymerase transient expression system relying on the bacteriophage T7 RNA polymerase with several key advantages was widely used for protein expression. VP6-encoding cDNA of rotavirus strain TB-Chen was cloned and constructed by inserting into prokaryotic expression plasmid pETL between bacteriophage T7 promoter and terminator regions. When eukaryotic cells were transfected with the recombinant plasmid containing the VP6 gene and infected with recombinant vaccinia virus vTF7-3 which provides bacteriophage T7 RNA polymerase, the VP6 gene was expressed. The findings showed that cytopathic effects by vaccinia virus infection were fast and severe leading to low level of protein expression. If cytopathic effects of the vaccinia virus can be reduced, higher level of the protein might be obtained.The findings provided a helpful means to further study the VP6 function and utilize the vaccinia virus/T7 RNA polymerase transient expression system.

Key words： Rotavirus VP6 Prokaryotic plasmid expression Eukaryotic cells Recombinant vaccinia virus vTF7-3 T7 RNA

Received: 07 December 2010 Published: 27 May 2011

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PAN Xiao-xia, ZHANG Shun, WEN Yu-ling, YUAN Jing, CHEN Yuan-ding. Expression of Prokaryotic Gene in Mammalian Cells Based on Recombinant Vaccinia Virus Encoding T7 RNA Polymerase. China Biotechnology, 2011, 31(5): 48-54.

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https://manu60.magtech.com.cn/biotech/ OR https://manu60.magtech.com.cn/biotech/Y2011/V31/I5/48

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[1]	. Expression of Prokaryotic Gene in Mammalian Cells Based on Recombinant Vaccinia Virus Encoding T7 RNA Polymerase[J]. China Biotechnology, 2011, 31(05): 0-0.

[2]	ZHANG Jing, LIU Huan, ZHOU Jing, LIU Jian-Hua. Large Protein Production Through Split Intein-Mediated Trans-Splicing[J]. China Biotechnology, 2009, 29(12): 74-78.

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