[Construction of recombinant adenovirus using modified AdEasy system].
To simplify AdEasy system for better efficiency, we attempted some modifications on the original system. Specifically, the Pme -linearized plasmid pAdtrack was transformed into competent AdEasier-1 cells prepared from treatment with CaCl(2). The DNA contained in the identified recombinant plasmid was digested with Pac and transfected into 293 cells to package the adenovirus, followed by identification of the recombinant adenovirus by means of observation of green fluorescence protein expression under fluorescence microscope. Chemical transformation of the linearized plasmid into AdEasier-1 cells resulted in very high success rate (20/20) for producing positive recombinant adenovirus clones, confirming the efficiency of the modified AdEasy system in constructing recombinant adenovirus.
Tang
Fu-xiang
FX
Department of General Surgery, Zhujiang Hospital, First Military Medical University, Guangzhou 510282, China.
Zheng
Quan
Q
Huang
Zong-hai
ZH
Zhang
Qing-guo
QG
Che
Xiao-yan
XY
D000256:Adenoviridae / Q000235:genetics*; D005822:Genetic Vectors; D006801:Humans; D010957:Plasmids / Q000235:genetics*; D011995:Recombination, Genetic; D014162:Transfection