TOXICOLOGICAL SIGNIFICANCE OF DOG LIVER CYTOCHROME-P-450 - EXAMINATION WITH THE ENZYME EXPRESSED IN SACCHAROMYCES-CEREVISIAE USING RECOMBINANT EXPRESSION PLASMID

被引：7

作者：

FUKUTA, H ^{[1
]}

OHI, H ^{[1
]}

UCHIDA, T ^{[1
]}

KOMORI, M ^{[1
]}

KITADA, M ^{[1
]}

KAMATAKI, T ^{[1
]}

机构：

[1] HOKKAIDO UNIV,FAC PHARMACEUT SCI,DIV ANALYT BIOCHEM,SAPPORO,HOKKAIDO 060,JAPAN

来源：

MUTATION RESEARCH | 1992年 / 269卷 / 01期

关键词：

DOG P-450IA1; EXPRESSION IN YEAST; MUTAGENIC ACTIVATION; IQ; MEIQ;

D O I：

10.1016/0027-5107(92)90164-W

中图分类号：

Q3 [遗传学];

学科分类号：

071007 ; 090102 ;

摘要：

A complementary DNA (cDNA) coding for a form of beagle dog cytochrome P-450 (Dah1), which is the orthologue to the CYP1A1 cDNA of rat, mouse and human, was inserted between the alcohol dehydrogenase (ADH) promoter and terminator regions of the yeast expression vector pAAH5. On introduction of the resulting recombinant plasmid pDC-1, Saccharomyces cerevisiae strain AH22 cells synthesized up to 1.5 x 10(5) molecules per cell of cytochrome P-450 protein (P-450(Dah1)). The carbon monoxide-bound reduced form of P-450(Dah1) showed an absorption peak at 447 nm and specific content of P-450(Dah1) was about 0.1 nmole P-450 per mg of microsomal protein. P-450(Dah1) cross-reacted with antibodies to rat P-448-H (CYP1A2) and dog P-450-D2 (CYP1A2). P-450(Dah1) activated 2-amino-3-methyl-imidazo[4,5-f]quinoline (IQ) and 2-amino-3,4-dimethylimidazo[4,5-f]quinoline (MeIQ) most efficiently in the umu test and exhibited a high activity of aryl hydrocarbon hydroxylase toward benzo[a]pyrene.

引用

页码：97 / 105

页数：9

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