Chinese Agricultural Science Bulletin ›› 2012, Vol. 28 ›› Issue (14): 110-113.doi: 10.11924/j.issn.1000-6850.2011-2249
Special Issue: 生物技术
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Abstract:
Extract the RNA of FMDV tape A from frozen cells in the laboratory,then acquire the cDNA by RT-PCR. A special primer pair was designed which containing BamHⅠand HindⅢ according to complete genome of FMDV,the target gene VP1 was amplified by PCR and subcloned into pMD 18-T vector.Then, digest the VP1 from pMD18-T vector and expression vector PET32a with BamHⅠand HindⅢ, ligated gene VP1 into PET32a and named recombinant plasmid PET32-VP1 after identification. The interest gene was induced to express in E.coli with IPTG and identified with SDS-PAGE.The target protein was purified with Ni-NTA Purification System. Result showed that the target protein was expressed in E.coli and also purified.This provide laboratory important material for further study.
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URL: https://www.casb.org.cn/EN/10.11924/j.issn.1000-6850.2011-2249
https://www.casb.org.cn/EN/Y2012/V28/I14/110