Construction of the Expression Vector of Viruslike Particles Containing FMDV IRES RNA
- VernacularTitle:含口蹄疫病毒IRES RNA病毒样颗粒表达载体的构建
- Author:
Min DOU
;
GuoGuang ZHANG
;
GuangFu YU
;
HongXin ZHANG
;
MingShan SHEN
;
Liang CHEN
;
- Publication Type:Journal Article
- Keywords:
MS2 bacteriophage Coat protein RNA virus Viruslike particles
- From:
China Biotechnology
2006;0(09):-
- CountryChina
- Language:Chinese
-
Abstract:
The Coat protein and Maturase gene of E.coli bacteriophage MS2 was amplified by PCR,then the gene was cloned into pET32a to construct the intermediate vector pET32aCP.The conservative sequence of FMDV internal ribosome entry site(IRES) was cloned into the downstream of pET32aCP bacteriophage gene to construct the prokaryotic expression vector pCPES.The recombinant plasmid pCPES transformed into E.coli strain BL21(DE3) was induced to express with 1mmol/L IPTG.The expression products were purified by sucrose density gradient centrifugation.The expression products observed by TEM were circular viruslike particles,and the diameter of these particles was about 26nm.The stability of viruslike particles was detected,and the viruslike particles was identified by RTPCR.The results showed that the viruslike particles contain the FMDV IRES RNA and have good stability.The viruslike particles have great prospect as the standard and quality control in the area of RNA virus detection.
