Construction of Eukaryotic Expressing Vector of FKBP12 and Establishment of Stable Transfectant A549 Cell Line

Objective: To construct the eukaryotic plasmid of FK506 Binding proteins 12 (FKBP12)and transfect A549 cell so as to establish stable cell line. Methods: The whole objective gene was cloned by RT-PCR from VSMCs. Eukaryotic victor pcDNA3.1/Hygro(+)-FKBP12 was connected and by enzyme digestion and DNA sequencing . We transfected the recombinant vector into A549 cell by lipofectamine TM 2000.Stable transfected A549 cell line was established after screening culture by Hygromycin B and was identified by Western blot. Results: The eukaryotic expression vector pcDNA3.1/Hygro(+)FKBP12 was constructed, stable transfected A549 cell line was established and FKBP12 protein was expressed successfully. Conclusion: The construction of the eukaryotic expression vector pcDNA3.1/Hygro (+)FKBP12 and the establishment of stable transfected A549 cell line not only lay the foundation for the further research into the mechanism of drug targets based on FKBP12 but also provide a safe and efficient way for myocardial new immunosuppressive agents.