Molecular cloning of cDNA encoding human lanosterol synthase

Biol Pharm Bull. 1995 Oct;18(10):1459-61. doi: 10.1248/bpb.18.1459.

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.

Publication types

  • Comparative Study

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Cloning, Molecular*
  • DNA Primers
  • DNA, Complementary / genetics
  • Humans
  • Intramolecular Transferases*
  • Isomerases / biosynthesis
  • Isomerases / genetics*
  • Molecular Sequence Data
  • Polymerase Chain Reaction
  • Sequence Homology, Amino Acid

Substances

  • DNA Primers
  • DNA, Complementary
  • Isomerases
  • Intramolecular Transferases
  • lanosterol synthase

Associated data

  • GENBANK/S81221