Analysis of endogenous S1P and LPA receptor expression in CHO-K1 cells

Gene. 2005 Apr 25;350(1):59-63. doi: 10.1016/j.gene.2005.01.016.

Abstract

The CHO-K1 cell line is commonly used for studies of recombinantly expressed proteins, including proteins of the G protein-coupled receptor (GPCR) family. This laboratory has used CHO-K1 cells for the functional characterization of Edg family GPCRs. However, parental CHO-K1 cells respond to lysophospholipids in in-vitro functional assays, which suggests expression of endogenous Edg family GPCRs. To determine the repertoire of Edg family receptor expression in this cell line, alignments of human and rodent sphingosine-1-phosphate (S1P) and lysophosphatidic acid (LPA) receptor sequences were used to design semi-redundant oligonucleotide pairs. A portion of each receptor gene coding sequence was amplified from Chinese hamster genomic DNA and the resultant gene fragments sequenced. Species-specific oligonucleotide pairs were designed using this novel sequence information and used to detect expression of S1P(1,2,4) and LPA(1) transcripts in CHO-K1 cells by RT-PCR.

Publication types

  • Comparative Study

MeSH terms

  • Animals
  • CHO Cells
  • Cricetinae
  • Cricetulus
  • DNA, Complementary / chemistry
  • DNA, Complementary / genetics
  • Gene Expression / genetics*
  • Humans
  • Mice
  • Molecular Sequence Data
  • Protein Isoforms / genetics
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Rats
  • Receptors, G-Protein-Coupled / genetics
  • Receptors, Lysophosphatidic Acid / genetics*
  • Receptors, Lysosphingolipid / genetics*
  • Reverse Transcriptase Polymerase Chain Reaction / methods
  • Sequence Analysis, DNA

Substances

  • DNA, Complementary
  • Protein Isoforms
  • RNA, Messenger
  • Receptors, G-Protein-Coupled
  • Receptors, Lysophosphatidic Acid
  • Receptors, Lysosphingolipid

Associated data

  • GENBANK/AY522543
  • GENBANK/AY522544
  • GENBANK/AY522545
  • GENBANK/AY522546
  • GENBANK/AY522547
  • GENBANK/AY522548
  • GENBANK/AY522549