Title: Construction of prokaryotic expression vector for HSV_1-TK and detection of expressed protein
Abstract: This paper discussed the self-destruction gene HSV1-TK prokaryotic expression in order to establish the foundation of bacteria vector applied in the tumor gene therapy.Firstly,plasmid pHSV-106 was taken as the template,the clone of HSV1-TK gene was acquired through PCR and connected to the prokaryotic expression vector pGEX-6P-1,then recombinant plasmid was identified and transformed into E.coli BL21(DE3) competence cell to observe the protein expression.The result showed that the length of TK gene was 1 128 bp,which was coincident to the theoretic value,and the identity between TK gene and the segment inserted into the vector pGEX-6P-1 was 100%;TK enzyme which was about 41 ku could be observed obviously through SDS-PAGE,and the amount increased with time.Experiment showed that the thymidine kinase from the herpes simplex virus could be expressed successfully in prokaryotic expression system;this result is the premise for the tumor gene therapy applying the bacteria as the vector that carried the sucide gene TK.
Publication Year: 2009
Publication Date: 2009-01-01
Language: en
Type: article
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