A quantitative method for cDNA-directed expression in cultured mammalian cells.
A simple technique of quantitative cDNA-directed expression was devised by using a vaccinia virus system. The technique was applied to the cDNA expression of peroxisomal acyl-coenzyme A oxidase (AOX) and microsomal cytochromes P450 1A2, 2E1, and 3A4. Quantitative cDNA expression of all four enzymes tested was successfully achieved. This represents important progress for establishing the relationship between the amount of enzyme and its function in cells. The vaccinia virus system was also used to express AOX protein in specific cellular regions.
Aoyama
T
T
Department of Biochemistry, Shinshu University School of Medicine, Nagano, Japan.
Nakajima-Nasu
T
T
Souri
M
M
Hashimoto
T
T
D042962:Acyl-CoA Oxidase; D000818:Animals; D015153:Blotting, Western; D002273:Carcinogens / Q000378:metabolism; D002460:Cell Line; D002875:Chromosomes / Q000378:metabolism; D003577:Cytochrome P-450 Enzyme System / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism; D018076:DNA, Complementary; D005786:Gene Expression Regulation; D005822:Genetic Vectors; D006801:Humans; D008830:Microbodies / Q000201:enzymology; D008861:Microsomes / Q000201:enzymology; D010088:Oxidoreductases / Q000096:biosynthesis / Q000378:metabolism; D010171:Palmitoyl Coenzyme A / Q000378:metabolism; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics; D013739:Testosterone / Q000378:metabolism; D014616:Vaccinia virus / Q000235:genetics*
D002273:Carcinogens; D018076:DNA, Complementary; D011993:Recombinant Fusion Proteins; D010171:Palmitoyl Coenzyme A; D013739:Testosterone; D003577:Cytochrome P-450 Enzyme System; D010088:Oxidoreductases; D042962:Acyl-CoA Oxidase