Construction of recombinant adenovirus containing human IL-24 gene and expression in A549 cells / 第三军医大学学报

ABSTRACT

Objective To construct the recombinant adenovirus encoding human IL-24 gene for future gene therapy. Methods The human IL-24 gene fragment was cloned into the shuttle plasmid pAdTrack-CMV to form the transfer vector by the method of homogenous recombination in bacteria. Then the recombinant adenovirus was transfected into 293T cells using Lipofectine DOTAP. The target gene was detected by polymerase chain reaction (PCR). The titer and its infection rate were determined using the green fluorescent protein (GFP) expression in the shuttle plasmid. The expression of target protein was measured by the method of immunohistochemistry. Results Restriction endonuclease and PCR analysis confirmed that the human IL-24 gene was successfully inserted into the adenovirus vector. The titer of the recombinant adenovirus was 1.2?10 10 pfu/ml. The adenovirus has a strong effect on A549 cells and human IL-24 can express in it. Conclusion The recombinant adenovirus containing human IL-24 gene was successfully constructed by the method of homogenous recombination in bacteria.