[Construction of the retroviral vector carrying the green fluorescent protein gene and its application in mediating gene transfer into T cells].
To construct retroviral vector carrying green fluorescent protein (GFP) gene, and determine whether T cells can be infected by retrovirus, the phosphoglycerate kinase (PGK) promoter gene and GFP full-length cDNA were subcloned into the retroviral vector pLXSN. The recombinant vector was transfected into PA317 packaging cells by DNA-calcium phosphate coprecipitation. The G418 resistant clones were selected, then NIH3T3 cells and T cells were infected by the culture supernatant of the retrovirus containing GFP. The results showed that GFP expression in packaging cell line PA317 was detectable under fluorescence microscope, which demonstrated that the GFP retrovirus were able to infect T cells. It is concluded the retrovirus vectors can introduce a foreign gene into T cells efficiently and can be used as important tools to deliver gene into T cells.
Chen
Xiang-Mei
XM
Department of Hematology, the Affiliated Hospital of Xuzhou Medical College, Xuzhou 221002, China. cxm95950@sina.com
Xu
Kai-Lin
KL
Pan
Xiu-Ying
XY
Li
Zhen-Yu
ZY
Lu
Qun-Xian
QX
Li
De-Peng
DP
D000818:Animals; D002460:Cell Line; D002478:Cells, Cultured; D005822:Genetic Vectors / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D008856:Microscopy, Fluorescence; D041681:NIH 3T3 Cells; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012190:Retroviridae / Q000235:genetics*; D013601:T-Lymphocytes / Q000166:cytology / Q000378:metabolism*; D014162:Transfection / Q000379:methods
D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins