[An easier method for generating recombinant adenovirus containing CD/TK fusion gene driven by vascular endothelial growth factor promoter].
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.


CONCLUSIONS
The improved homologous recombination in bacteria is efficient, convenient and easy to be carried out.
Huang
Yuan-yuan
YY
Department of General Surgery, Zhujiang Hospital, First Military Medical University, Guangzhou 510282, China.
Huang
Zong-hai
ZH
Chen
Jian-fa
JF
Tang
Fu-xiang
FX
Yang
Wen-yu
WY
Zheng
Quan
Q
Che
Xiao-yan
XY
D000256:Adenoviridae / Q000235:genetics; D043525:Cytosine Deaminase / Q000235:genetics*; D010957:Plasmids; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000235:genetics*; D011995:Recombination, Genetic; D013937:Thymidine Kinase / Q000235:genetics*; D014162:Transfection; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics*
D011993:Recombinant Fusion Proteins; D042461:Vascular Endothelial Growth Factor A; D013937:Thymidine Kinase; D043525:Cytosine Deaminase