Expression of Mycobacterium Tuberculosis PE35 in 293T cells

Objective To construct the eukaryotic expression vector of pcDNA-FLAG-PE35 and evaluate Mycobacterium Tuberculosis(TB) PE35 expression in SV40-transformed embryonic kidney cell line 293T.Methods The full-length coding gene of Mycobacterium Tuberculosis PE35 was amplified from a plasmid with standard PCR method.The expression construct of pcDNA-FLAG-PE35 was generated by inserting the amplified gene of TB PE35 into the eukaryotic expression vector of pcDNA3-FLAG.The expressed protein of TB PE35 was evaluated by Western blot in transient transfected 293T cells.Results The pattern of restriction enzyme digestion and result of nucleotide sequencing confirmed that the gene of TB PE35 was accurately inserted into eukaryotic expression vector pcDNA-FLAG-PE35.The expression of recombinant protein was detected by Western blot.Conclusion The production of recombinant protein will facilitate the understanding of molecular mechanism of TB PE35 related pathogenesis in TB infection.