Related Experiment Video
Updated: Jul 14, 2026
Generation of Plasmid Vectors Expressing FLAG-tagged Proteins Under the Regulation of Human Elongation Factor-1α Promoter Using Gibson Assembly
Published on: February 9, 2015
[Construction of CYP1A1-Wt and Asn461, Val462 gene eukaryotic expression vector and itstransfection in HLF cells]
Yongmei Xiao1, Qing Wei, Yimin Liu
1Department of Preventive Medicine, School of Public Health, Sun Yatsen University, Guangzhou 510080, China.
Objective:
To establish the cells model with high expression CYP1A1 and analyze the expression of the variable genotype.
Methods:
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.
Results:
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.
Conclusion:
HLF cells with wild-CYP1A1 or variant-CYP1A1 ware established successfully.
