Molecular cloning of cDNA encoding rat 2,3-oxidosqualene: lanosterol cyclase.
A cDNA encoding rat 2,3-oxidosqualene: lanosterol cyclase, the enzyme responsible for the complex cyclization/rearrangement step in sterol biosynthesis, was cloned by extensive application of PCRs. Oligonucleotide primers were designed in sense and anti-sense directions corresponding to internal peptide sequences of purified enzyme. Successive PCRs with all possible combinations of these primers yielded a 178-bp fragment, and based on its sequence full nucleotide sequence of cDNA was obtained by a "rapid amplification of cDNA ends" (RACE) method.
Kusano
M
M
Faculty of Pharmaceutical Sciences, University of Tokyo, Japan.
Shibuya
M
M
Sankawa
U
U
Ebizuka
Y
Y
D000595:Amino Acid Sequence; D000818:Animals; D017360:Arabidopsis / Q000378:metabolism; D001483:Base Sequence; D002176:Candida albicans / Q000378:metabolism; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000096:biosynthesis* / Q000737:chemistry; D019751:Intramolecular Transferases; D007535:Isomerases / Q000096:biosynthesis*; D008099:Liver / Q000378:metabolism; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D051381:Rats; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D014158:Transcription, Genetic
D018076:DNA, Complementary; D007535:Isomerases; D019751:Intramolecular Transferases; C021924:lanosterol synthase