2012•Journal of Southeast UniversityRequires access
Construction of pGEX-5X-1-hLMO4 prokaryotic plasmid and identification of its recombinant protein
Feng Li
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Abstract
Objective:To construct prokaryotic expression vector of human LMO4 gene and induce,purify and identify its recombinant protein expression.Methods:The hLMO4 coding sequence was digested with BamHⅠ and XhoⅠ enzymes,and cloned into pGEX-5X-1.The expression of GST-hLMO4 fusion protein was induced by IPTG and identified by Western blot.Results:The coding sequence of hLMO4 gene was cloned into the pGEX-5X-1 plasmid which was transformed into E.coli BL21.The length of fragment was 500 bp.The expression of GST-hLMO4 fusion protein was induced by IPTG,and the molecular weight of protein was 49 000 Da.Conclusion:The recombinant prokaryotic plasmid was successfully constructed into pGEX-5X-1.The expression of GST-LMO4 fusion protein was induced by IPTG and identified.