PPAR-alpha cloning, expression, and characterization

Methods Mol Biol. 2013:952:7-34. doi: 10.1007/978-1-62703-155-4_2.

Abstract

Peroxisome proliferator-activated receptor α (PPARα) is a member of the nuclear/steroid receptor gene superfamily that also comprises β, δ, and γ isoforms. PPARα is a ligand-activated transcription factor that plays an important role in the regulation of many genes involved in key metabolic processes. Today, PPARα has been cloned from mammalian, marsupial, and a number of marine species and its expression has been found to be relatively tissue- and species-specific. Here, we describe the methods for cloning of PPARα genes by RT-PCR and RACE approaches and related protocols for studying the expression of cloned PPARα cDNAs in mammalian cell systems.

MeSH terms

  • Animals
  • COS Cells
  • Cell Nucleus / metabolism
  • Chlorocebus aethiops
  • Cloning, Molecular / methods*
  • Cytosol / metabolism
  • DNA, Complementary / biosynthesis
  • DNA, Complementary / genetics
  • DNA, Complementary / isolation & purification
  • Gene Expression Regulation*
  • Humans
  • Mice
  • PPAR alpha / genetics*
  • Plasmids / genetics
  • RNA, Messenger / genetics
  • RNA, Messenger / isolation & purification
  • Rats
  • Reverse Transcriptase Polymerase Chain Reaction
  • Sequence Analysis

Substances

  • DNA, Complementary
  • PPAR alpha
  • RNA, Messenger