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MOLECULAR CLONING OF cDNA ENCODING HUMANLANOSTEROL SYNTHASE

ChungKi SUNG, Masaaki SHIBUYA, Ushio SANKAWA, Yutaka EBIZUKA

Author information

ChungKi SUNG
Masaaki SHIBUYA
Ushio SANKAWA
Yutaka EBIZUKA
Author's Organization：
Faculty of Pharmaceutical Sciences, The University of Tokyo:(Present address)College of Pharmacy, Chonnam National University
Faculty of Pharmaceutical Sciences, The University of Tokyo
Faculty of Pharmaceutical Sciences, The University of Tokyo:(Present address)Faculty of Pharmaceutical Sciences, Toyama Medical & Pharmaceutical University
Faculty of Pharmaceutical Sciences, The University of Tokyo

Corresponding author

Keywords:lanosterol synthase,cDNA,human liver,PCR,2, 3-oxidosqualene,lanosterol,cyclase

JOURNALFREE ACCESS

1995 Volume 18Issue 10Pages 1459-1461

DOIhttps://doi.org/10.1248/bpb.18.1459

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Published: October 15, 1995Received: August 17, 1995Available on J-STAGE: April 10, 2008Accepted: August 31, 1995Advance online publication: -Revised: -

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Article overview

Abstract

A cDNA encoding human lanosterol synthase, the enzyme responsible for the backbone formation step in sterol biosynthesis, was cloned by extensive application of PCRs. Five degenerate oligonucleotide primers (139S, 440S, 528A, 575A and 712A) corresponding to the homologous amino acid sequences among the known 2, 3-oxidosqualene cyclase (OSC) were designed. PCR with one pair (440S and 528A) of five primers yielded a 285-bp fragment. PCRs with the primers based on the obtained fragment and the degenerate primers (139S and 712A) gave longer fragments. Finally, full nucleotide sequence of cDNA was obtained by a "rapid amplification of cDNA ends"(RACE) method.

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