Two-step transformation for highly efficient construction of a replication-defective recombinant adenoviral vector.

Objective: To efficiently construct a replication-defective recombinant adenoviral vector using a two-step transformation procedure.

Methods: Plasmid pAdEasy-1 was linearized and transformed into E.coli BJ5183 to construct BJ5183pAdEasy-1 cells.

[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.

[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.

Effects of TNP-470 on proliferation and apoptosis in human colon cancer xenografts in nude mice.

Aim: To study the effect of TNP-470 on cell growth, proliferation and apoptosis in human colon cancer xenografts in nude mice.

Methods: Human colon cancer xenografts were transplanted into 20 nude mice. Mice were randomly divided into two groups.