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

Objective To construct the eukaryotic plasmid of Rho guanine nucleotide dissociation inhibitors-2(LyGDI) and transfect A549 cell so as to establish stable cell line.Methods The gene of LyGDI was amplified by PCR.Eukaryotic vector pEGFP-C1-LyGDI was constructed and confirmed by enzyme digestion、PCR、DNA sequencing.We transfected the recombinant vector into A549 cell by lipofectamineTM 2000.Stable transfected A549 cell line was established after screening culture by G418 and was identified by RT-PCR and Western blot.Results The eukaryotic expression vector pEGFP-C1-LyGDI was constructed,stable transfected A549 cell line was established and LyGDI protein was expressed successfully.Conclusion The construction of the eukaryotic expression vector pEGFP-C1-LyGDI and the establishment of stable transfected A549 cell line have laid the experiment foundation for further studies on the effect of over expression on invasion and metastasis of cancer.