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Open Access Green as soon as Postprint is submitted to ZB.
Genetically engineered V79 Chinese hamster cells for stable expression of human cytochrome P450IA2.
Eur. J. Pharmacol. 228, 95-102 (1992)
V79 Chinese hamster cells were genetically engineered for stable expression of human P450IA2. Full length cDNA, encoding human P450IA2, was inserted into an SV40 early promoter containing eukaryotic expression vector and cointroduced with the selection marker neomycin phosphotransferase (conferring resistance to the neomycin derivative G418) into V79 Chinese hamster cells. The recombinant expression vector was introduced into two different V79 sublines, one expressing an endogenous acetyltransferase (V79-NH), the other not (V79-MZ). The presence of human cytochrome CYP1A2 cDNA in the G418 resistant V79 cell clones was confirmed by Southern blotting. The transcription of the cDNA into mRNA was detected by Northern blotting and the translation into an authentic cytochrome P450IA protein was shown by Western blotting. The enzymatic activity in these cells was determined by the cytochrome P450IA2-dependent methoxy-, ethocy-, and pentoxyresooruffa dealkylation activity.
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Publication type
Article: Journal article
Document type
Scientific Article
Keywords
Cyp1a2 Recombinant Expression Vector ; Cytochrome P450ia2 (human) ; Heterologous Expression ; P450-dependent Enzymatic Activity ; V79 Chinese Hamster Cells
ISSN (print) / ISBN
0014-2999
e-ISSN
0014-2999
Journal
European Journal of Pharmacology
Quellenangaben
Volume: 228,
Issue: 2-3,
Pages: 95-102
Publisher
Elsevier
Non-patent literature
Publications
Reviewing status
Peer reviewed