[Construction of recombinant adenovirus vector carrying secondary lymphoid organ chemokine]

Ping Chen, Ling-lin Yang, Huo-zhen Hu
Sichuan da Xue Xue Bao. Yi Xue Ban, Journal of Sichuan University. Medical Science Edition 2007, 38 (3): 365-9

OBJECTIVE: To construct a recombinant adenovirus vector carring the SLC gene for further studies on gene therapy for carcinoma.

METHODS: The SLC gene was amplified from the pORF5 vector with polymerase chain reaction (PCR) technique. The amplified gene was cloned into the pENTR11 vector. With the pENTR11-SLC plasmid and the backbone plasmid pAd/CMV/V5-DEST, the homologous recombination reaction took place in vitro. The reaction mixture was transferred into TOP10 E. coli strains (without F' episome). The recombination adenovirus plasmid was then generated. The recombinant adenoviruses were packaged and amplified in 293A cells.

RESULTS: The SLC gene was successfully cloned into the pAd/CMV/V5-DEST plasmid and the recombinant adenoviruses carrying SLC gene were detected by PCR, with a viral titer of 2. 6 X 10(8) pfu/mL.

CONCLUSION: The constructed recombinant adenovirus can introduce SLC gene into tumor tissues, which provides a foundation for the study of antitumor efficacy of SLC.

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