Construction of CYP1A1-Wt and Asn461, Val462 gene eukaryotic expression vector and itstransfection in HLF cells
Xiao, Y.; Wei, Q.; Liu, Y.; Wang, H.
Wei Sheng Yan Jiu 36(2): 217-219
2007
ISSN/ISBN: 1000-8020 PMID: 17555106 Document Number: 606411
To establish the cells model with high expression CYP1A1 and analyze the expression of the variable genotype. Clone the target fragment (CYP1A1-Wt, Asn461, and -Val462) in T-Vectors into pBABE-neo, then transfer into the human embryonic lung fibroblast cells (HLF). The positive cells were selected by G418. The expression of CYP1A1 protein was measured by Western-Blot. At the same time, cells shape, growth of curve, and carcinogenicity were observed. HLF transferred with wild-CYP1A1 and variant-CYP1A1 expressed CYP1A1 protein higher than normal HLF. No differences of biology were found between the transfected HLF and normal HLF. These cells are not belong to carcinogenesis cells. HLF cells with wild-CYP1A1 or variant-CYP1A1 ware established successfully.