[Construction of CYP1A1-Wt and Asn461, Val462 gene eukaryotic expression vector and itstransfection in HLF cells].
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.


CONCLUSIONS
HLF cells with wild-CYP1A1 or variant-CYP1A1 ware established successfully.
Xiao
Yongmei
Y
Department of Preventive Medicine, School of Public Health, Sun Yatsen University, Guangzhou 510080, China.
Wei
Qing
Q
Liu
Yimin
Y
Wang
Hui
H
D000483:Alleles; D019363:Cytochrome P-450 CYP1A1 / Q000096:biosynthesis* / Q000235:genetics; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism*; D005822:Genetic Vectors; D006801:Humans; D008168:Lung / Q000166:cytology* / Q000196:embryology / Q000378:metabolism; D014162:Transfection
D019363:Cytochrome P-450 CYP1A1