Construction of the lentivirus vector over-expressing PCGF1 and establishment of A549 cell line stably over-expressing PCGF1

Abstract

Objective 　To construct lentiviral over-expression vector of polycomb group ring finger 1(PCGF1) and get the A549 cell stably over-expressing PCGF1. Methods 　The primers were designed according to human PCGF1 sequence. The target gene was amplified by PCR with the template of the A549 cDNA. The target gene was digested and then inserted into the pLVX-IRES-puro plasmid. After the lentiviral vector was packaged, A549 cell line infected by the packaged virus was selected by puromycin, and the expression of PCGF1 was verified by Western blotting. Results 　The DNA sequence of recombinant plasmid was verified to be correct, and then transfected into 293T cells. Western blotting showed the remarkably escalated expression of PCGF1 gene in 293T cells. After lentivirus infection, Western blotting supported that the PCGF1 was stably over-expressed in A549 cells after puromycin selection. Conclusion 　The lentiviral over-expression vector pLVX-IRES-PCGF1-puro has been successfully constructed, and the recombinant virus can effectively infect A549 cells and the PCGF1 is stably over-expressed.