Construction and Identification of Eukaryotic Expression Vector of Folate Binding Proteins 1

Objective To construct the eukaryotic expression vector of pcDNA3.1-folate binding proteins 1(Folbp1) and detect its expression in lung adenocarcinoma A549 cell line.Methods The complete coding sequence of Folbp1 gene was amplified by reverse transription-PCR from lung adenocarcinoma A549 cell and was cloned into eukaryotic expression vector pcDNATM 3.1/myc-His B.The recombinant plasmid was then transfected into A549 cell by liposome.After screening culture by G418,stable transfected A549 cell line was established.The expression of Folbp1 protein was identified by Western blot.Results The Folbp1 gene was amplified by RT-PCR and detected by SDS-PAGE,the expected 823 bp was identified.The pMD-Folbp1 ligation products were transformed into the competent bacterial cells and selected by ampicillin,the positive clones were gained.Folbp1 gene was amplified and ligated with pcDNATM 3.1/myc-His B expression vector,in the electrophoresis after restriction enzyme reaction;about 5.1 kb vector band and a specific band were obtained.The pcDNA 3.1-Folbp1 plasmid was transfected into A549 cell and screened by G418,the protein of expected size was identified by Western blot.Conclusions The eukaryotic expression vector of Folbp1 has been successfully established and the stably transfected A549 cell line also established,which pave the way for further studies on the function of Folbp1 in the development of lung.