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KMID : 0364819870250030165
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Korean Journal of Microbiology
1987 Volume.25 No. 3 p.165 ~ p.172
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Construction of an expression vector with SV40 DNA in a mammalian cell
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Chung, M.H/정민혜
Kim, S.H./Jun, H.S./Rho, H.M./김상해/전희숙/노현모
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Abstract
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1
An expression vector in a mammalian cell was constructed using the origin of replication (OR) and the promoters of SV40. The plasmid pSVOE was constructed by inserting SV40 DNA fragment (1,118 bp) containing SV40 OR and promoters into pBR322-1, and then a multiple cloning sequence was inserted at the immediate downstream of the late promoter of SV40 in the pSVOE vector. The plasmid was named pSVML. As a selection marker, thymidine kinase gene of herpes simplex virus with its promoter was inserted into EcoRI site of pSVML and the recombinant was named pSVMLTKp.
To test the ex-ression capacity of foreigen gene inserted at the multiple cloning site of pSVML, the thymidine kip se gene without its own promoter was inserted at the BamHI site of pSVML. The recombinant was named pSVML-TK.Tbese plasmids,,pSVML-TKp and pSVML-TK, were transfected into COS cells with calcium phosphate precipitation method. The thymidine kinase activity was significantly increased in both transfected cells.
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KEYWORD
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