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Construction of the expression vector of virus-like particles containing FMDV IRES RNA |
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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 pET32a-CP. The conservative sequence of FMDV internal ribosome entry site (IRES) was cloned into the downstream of pET32a-CP 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 virus-like particles, and the diameter of these particles was about 26 nm.. The stability of virus-like particles was detected, and the virus-like particles was identified by RT-PCR. The results showed that the virus-like particles contain the FMDV IRES RNA and have good stability. The study shows that the virus-like particles have great prospect as the standard and quality control in the area of RNA virus detection.
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Received: 14 May 2007
Published: 25 September 2007
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