[An easier method for generating recombinant adenovirus containing CD/TK fusion gene driven by vascular endothelial growth factor promoter].
作者信息
Huang Yuan-yuan, Huang Zong-hai, Chen Jian-fa, Tang Fu-xiang, Yang Wen-yu, Zheng Quan, Che Xiao-yan
机构信息
Department of General Surgery, Zhujiang Hospital, First Military Medical University, Guangzhou 510282, China.
出版信息
Di Yi Jun Yi Da Xue Xue Bao. 2004 Jan;24(1):117-20.
OBJECTIVE
To efficiently construct the recombinant adenovirus containing CD/TK fusion gene driven by vascular endothelial growth factor (VEGF) promoter using improved homologous recombination in bacteria.
METHODS
Chemical transformation, instead of electroporation, of the plasmid pAdEasy-1 into E.coli BJ5183 strain was performed to prepare BJ5183pAdEasy-1 as the competent bacterium, in which pAdEasy-1 and pAdtrack-VEGFP-CDglyTK were recombined. Finally, the recombinant adenovirus of AdVEGFP-CDglyTK was constructed by transfecting 293 cells with linearized adenoviral genomes of pAdEasy-VEGFP-CDglyTK.
RESULTS
This new transformation procedure generated recombinant plasmids in a yield of 60% (6/10), and the adenovirus AdVEGFP-CDglyTK was harvested 7-12 d after transfection.
CONCLUSION
The improved homologous recombination in bacteria is efficient, convenient and easy to be carried out.
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