Application of the random arbitrary primed polymerase chain reaction differential display method to isolate genes of cholesterol metabolism-related proteins from rat liver

Masao Sato, Susumu Yoshida, Koji Nagao, Shoko Nishizono, Mihoko Kusaba, Mei Chu Hung, Ikuo Ikeda, Katsumi Imaizumi

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

The random arbitrary primed (RAP) polymerase chain reaction (PCR) differential display (DD) method was applied to isolate genes related to cholesterol metabolism from exogenously hypercholesterolemic (ExHC) rats and the progenitor, SD rats. Forty-seven trials of RAP-PCR DD resulted in the isolation of 37 clones differing in strain, cholesterol supplementation or their interaction. Among their fingerprints, five clones gave reproducible patterns by a Northern blotting analysis. The sequence of two clones with lower mRNA abundance in ExHC rats than in SD rats was homologous to that of fatty acid synthase and oxalyl-CoA decarboxylase. Two other clones with higher mRNA on the ncholesterol diet were matrin F/G protein and the NMDA receptor glutamate-binding subunit. The other clone with higher mRNA abundance in ExHC rats on the cholesterol diet was myelodysplasai/myeloid leukemia factor 2. Fifteen trials of reverse transcriptase (RT)-PCR DD yielded 10 clones, but none of the fingerprints were reproduced by the Northern blotting analysis. These results indicate that RAP-PCR DD is an appropriate alternative to RT-PCR DD for isolating the genes involved in hypercholesterolemia.

Original languageEnglish
Pages (from-to)1058-1060
Number of pages3
JournalBioscience, Biotechnology and Biochemistry
Volume64
Issue number5
DOIs
Publication statusPublished - 2000 Jan 1
Externally publishedYes

Keywords

  • Differential display
  • Hypercholesterolemia
  • Random arbitrary primed polymerase chain reaction
  • Reverse transcriptase polymerase chain reaction

ASJC Scopus subject areas

  • Biotechnology
  • Analytical Chemistry
  • Biochemistry
  • Applied Microbiology and Biotechnology
  • Molecular Biology
  • Organic Chemistry

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