Genetically engineered V79 Chinese hamster cells for stable expression of human cytochrome P450IA2

Catherine Wölfel, Barbara Heinrich-Hirsch, Thomas Schulz-Schalge, Albrecht Seidel, Heinz Frank, Uwe Ramp, Felix Wächter, Friedrich J. Wiebel, Frank Gonzalez, Helmut Greim, Johannes Doehmer

Research output: Contribution to journalArticlepeer-review

59 Scopus citations

Abstract

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.

Original languageEnglish
Pages (from-to)95-102
Number of pages8
JournalEuropean Journal of Pharmacology: Environmental Toxicology and
Volume228
Issue number2-3
DOIs
StatePublished - 1 Sep 1992
Externally publishedYes

Keywords

  • CYP1A2 recombinant expression vector
  • Cytochrome P450IA2 (human)
  • Heterologous expression
  • P450-dependent enzymatic activity
  • V79 Chinese hamster cells

Fingerprint

Dive into the research topics of 'Genetically engineered V79 Chinese hamster cells for stable expression of human cytochrome P450IA2'. Together they form a unique fingerprint.

Cite this