Title: Construction of Prokaryotic Expression Plasmid Included HSV_1-TK Gene and Detection of Expressed Protein
Abstract: Objective:To construct the prokaryotic expression plasmid included HSV1-TK gene and detect the result of expression,the basis for using bacteria vector in tumor gene therapy was made in this work.Method:Plasmid pHSV-106 was taken as the template firstly,HSV1-TK gene was cloned by PCR and connected to the expression vector pBV220,then the recombinant plasmid was identified and transformed into E.coli DH5α competence cell,and the expressed protein would be detected.Result:The molecule weight of TK gene was 1 128bp which is equal to the size in theory,and the identity between TK gene(NCBI accession number: V00470) and the cloned segment is 100%;But the TK enzyme can't be observed by electrophoresis,even though many repeats and different hosts were tried,so influences induced by errors and rare codons could be ignored;TK enzyme which is about 41kD can be obviously detected in plasmid pET-28a-TK.Therefore,this study showed that the thymidine kinase from the herpes simplex virus can be expressed successfully in prokaryotic expression vector.Conclusion:The selection of prokaryotic expression system could be regarded as the premise to tumor gene therapy that hopes to apply bacteria as vector.
Publication Year: 2008
Publication Date: 2008-01-01
Language: en
Type: article
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