Isolation and identification of the gene of cholesterol oxidase from Brevibacterium sterolicum ATCC 21387, a widely used enzyme in clinical analysis.
The gene coding cholesterol oxidase (CHOD) from Brevibacterium sterolicum, which is widely used in clinical analysis, has been selected from pUC19-based gene bank in E. coli MM294 by colony-hybridization using synthetic DNA as probe. The gene was identified to encode the protein having the same amino acid sequence as that determined from amino-acid sequence analysis. The expression of the CHOD gene in E. coli was not observed, probably due to the transcription failure. Attempts are being made to express it in various hosts including Streptomyces lividans, Corynebacterium glutamicum, and B. sterolicum itself.
Fujishiro
K
K
Tokyo Research Laboratories, Kyowa Hakko Kogyo Co., Ltd., Japan.
Ohta
T
T
Hasegawa
M
M
Yamaguchi
K
K
Mizukami
T
T
Uwajima
T
T
Ota
T
T
D000595:Amino Acid Sequence; D001483:Base Sequence; D015139:Blotting, Southern; D001951:Brevibacterium / Q000201:enzymology / Q000235:genetics*; D002789:Cholesterol Oxidase / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular / Q000379:methods; D015342:DNA Probes; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification; D004926:Escherichia coli / Q000235:genetics; D015723:Gene Library; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D011994:Recombinant Proteins / Q000378:metabolism; D015183:Restriction Mapping; D012689:Sequence Homology, Nucleic Acid
D015342:DNA Probes; D004269:DNA, Bacterial; D015345:Oligonucleotide Probes; D011994:Recombinant Proteins; D002789:Cholesterol Oxidase