An easier method for generating recombinant adenovirus containing CD/TK fusi on gene driv-en by vascular endothelial growth factor promoter

Abstract

Abstract: Objective To efficiently construct the recombinant adenov ir us containing CD/TK fusion gene driven by vascular endothelial growth factor (V EGF) promoter using improved homologous recombination in bacteria.Methods Chemi cal trans-formation, instead of electroporation, of the plasmid pAdEasy-1 into E.coli BJ5183 strain was performed to prepare BJ5183pAdEasy-1 as t he competent bacterium, in which pAdEasy-1 and pAdtrack-VEGFP-CDglyTK were recom bined. Fi-nally, the recombinant adenovirus of AdVEGFP-CDglyTK was constructed b y tranfecting 293 cells with linearized adenoviral genomes of pAdEasy-VEGFP-CDg lyTK.Results This new transformation procedure generated recombinant plasmids i n a yield of 60% (6/10), and the adenovirus AdVEGFP-CDglyTK was harvested 7-12 d after transfection.Conclusion The im-proved homologous recombination in bacte ria is efficient, convenient and easy to be carried out.

CLC Number:

Q784
R373.1

HUANG Yuan-yuan¹, HUANG Zong-hai¹, CHEN Jian-fa¹, TANG Fu-xiang¹, YANG Wen-yu¹, ZHENG Quan¹, CHE Xiao-yan². An easier method for generating recombinant adenovirus containing CD/TK fusi on gene driv-en by vascular endothelial growth factor promoter[J]. Journal of Southern Medical University, 2004, 24(01): 117-120.

References

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