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- W2374818024 abstract "Objective: To construct the replication-deficient recombinant adenovirus containing human mindkine promoter and thymidine kinase gene of Herpes simplex virus(HSV-TK).Methods: Based on Adeno-X~(TM) expression system,CMV promoter of pShuttle was replaced by MK promoter.The SHV TK gene,which was obtained from plasmid p HSV-106,was subcloned into p Shuttle vector under MK promoter.The proper p Shuttle-MK-TK was identified by special restriction endonuclease analysis.Both of the plasmid p Shuttle-MK-TK and Adeno-X DNA were digested with restriction endonuclease PI-SceI and I-CeuI,the target fragment(MK+TK) was subcloned into the Adeno-X DNA with ligation in vitro and the recombinant Adeno-MK-TK was obtained.After linearized by digesting with restriction endonucleases PacI,the plasmid Adeno-MK-TK was transfected into HEK 293 to produce recombinant virus stocks.Using PCR method,the virus stocks was identified whether it contained the interest gene MK and TK.Results: The restriction endonuclease analysis showed that the MK promoter and TK gene were inserted into the pShuttle successfully.And,the TK gene was located on the downstream of the MK promoter.PCR products were in coincidence with expectedness.Conclusion: By ligation in vitro,the recombinant HSV-TK gene under MK promoter mediated by adenovirus vector was constructed successfully." @default.
- W2374818024 created "2016-06-24" @default.
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- W2374818024 date "2006-01-01" @default.
- W2374818024 modified "2023-09-26" @default.
- W2374818024 title "Construction of replication recombinant adenoviruscontaining human mindkine promoter and HSV-TK" @default.
- W2374818024 hasPublicationYear "2006" @default.
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