A Primary Study on the Constructed GFP Lentivirus Vector's Transient Expression Kinetics in the Chicken Embryo Cells by Liposome-mediating

GFP is a reporter gene by which gene expressing and location of protein can be detected. The pLenti6/V5-D-TOPO is a lentiviral expressing vector with high efficiency. In this expriment, the GFP gene was amplified by PCR from the plasmid pEGFP-N1, then it was ligated into the pLenti6/V5-D-TOPO by the TOPO cloning efficiently, quickly and directly, and constructed the recombined pLEGFP vector that could transfect the cells efficently and display protein expressing expediently. The results of PCR and transient transfecting of the pLEGFP testified that the GFP gene was ligated correctly into the pLenti6/V5-D-TOPO and that the pLEGFP could express transiently in the Hela cells and the chicken embryo cells at stage X. But transient transfection effect has an obvious difference between recombinant plasmid and control plasmid. The results of research show that the molecular size and the ratio of plasmid and liposome have the key effect on transfection. This work also provides a tool for transgenic chicken research.