Construction of a Chimeric Adenovirus Vector with High Infection Power and Detection of its Infection Ability by Flow Cytometry

Objective To construct a chimeric adenovirus vector which is a fiber-substituted adenovirus type 5 (Ad5) vector containing adenovirus type 35 (Ad35) fiber,and to investigate its infection ability in cancer cells by flow cytometry.Methods The Fiber sequence of Ad35 was amplified using PCR,then inserted and replaced the Fiber sequence of Ad5 to generate a chimeric adenovirus vector Ad5-F35.Ad5-F35 was loaded the enhanced green fluorescent protein gene (EGFP) to generate Ad5-F35EGFP.Ad5-F35EGFP was used to infect the raji lymphoma cells and SGC-7901 gastric cancer cells at an MOI of 5 for the purpose of detecting EGFP expression and judging infection ability of this chimeric adenovirus vector by flow cytometry.Results The chimeric adenovirus vector Ad5-F35EGFP expressing EGFP was successfully constructed and recombined with the virus titer of 3.4×109 pfu/ml.The flow cytometry found that Ad5-F35EGFP expressed EGFP with higher efficiency in raji lymphoma cells and SGC-7901 gastric cancer cells compared with the traditional vector Ad5-EGFP.Conclusion The chimeric adenovirus vector Ad5-F35 has a higher infection power in cancer cells,which is important for increasing the transgene transfer ability of adenovirus vector and improving the gene therapeutic efficacy of clinical cancer treatment.