Cloning of human cytochrome P-450 cDNA and its expression in Saccharomyces cerevisiae.
A human cytochrome P-450 cDNA clone, lambda hPA6, was isolated from a human adult liver cDNA library. The expression plasmid pHM3 delta 2 was constructed by insertion of lambda hPA6 and a synthetic DNA into the yeast mRNA and reduced carbon monoxide-difference spectra of the yeast microsomes revealed that only the yeast transformed by the expression plasmid pHM3 delta 2 produced foreign cytochrome P-450. In addition, the expressed protein reacted with antiserum to P-450-HM2, a form probably identical with P-450MP, and showed a detectable level of mephenytoin 4-hydroxylase activity.
Ohgiya
S
S
Government Industrial Development Laboratory, Hokkaido, Agency of Industrial Science and Technology, Sapporo, Japan.
Komori
M
M
Fujitani
T
T
Miura
T
T
Shinriki
N
N
Kamataki
T
T
D000595:Amino Acid Sequence; D001189:Aryl Hydrocarbon Hydroxylases; D001483:Base Sequence; D015153:Blotting, Western; D002851:Chromatography, High Pressure Liquid; D003001:Cloning, Molecular; D065731:Cytochrome P-450 CYP2C19; D003577:Cytochrome P-450 Enzyme System / Q000096:biosynthesis / Q000235:genetics*; D005822:Genetic Vectors; D006801:Humans; D008099:Liver / Q000032:analysis; D008861:Microsomes / Q000201:enzymology; D006899:Mixed Function Oxygenases / Q000235:genetics; D008969:Molecular Sequence Data; D010957:Plasmids; D012441:Saccharomyces cerevisiae / Q000235:genetics*
D003577:Cytochrome P-450 Enzyme System; D006899:Mixed Function Oxygenases; D001189:Aryl Hydrocarbon Hydroxylases; C045793:CYP2C19 protein, human; D065731:Cytochrome P-450 CYP2C19