Construction of eukaryotic expressing vector of guanine nucleotide dissociation inhibitor-2 and establishment of stable transfectant A549 cell line

Objective To construct the eukaryotic plasmid of guanine nucleotide dissociation inhibitors-2(GDI2) and transfect A549 cell so as to establish stable cell line.Methods The gene of GDI2 was amplified by polymerase chain reaction(PCR).Eukaryotic vector pcDNA3.1-GDI2 was constructed and confirmed by enzyme digestion,PCR and DNA sequencing.The recombinant vector was transfected into A549 cell by lipofectamine TM 2000.Stable transfected A549 cell line was established after screening culture by G418 and was identified by Western blot method.Results The eukaryotic expression vector pcDNA3.1-GDI2 was constructed,stable transfected A549 cell line was established,and GDI2 protein was expressed successfully.Conclusion The successful construction of the eukaryotic expression vector pcDNA3.1-GDI2 and the establishment of stable transfected A549 cell line have laid the experiment foundation for further studies on invasion and metastasis of cancer.