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

被引：19

作者：

Holdsworth, G

Slocombe, P

Hutchinson, G

Milligan, G

机构：

[1] Celltech Ltd, Div Biol, Slough SL1 4EN, Berks, England

[2] Univ Glasgow, Mol Pharmacol Grp, Div Biochem & Mol Biol, Inst Biomed & Life Sci, Glasgow G12 8QQ, Lanark, Scotland

来源：

GENE | 2005年 / 350卷 / 01期

关键词：

Chinese hamster; sphingosine-1-phosphate; lysophosphatidic acid; RT-PCR;

D O I：

10.1016/j.gene.2005.01.016

中图分类号：

Q3 [遗传学];

学科分类号：

071007 ; 090102 ;

摘要：

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-I-phosphate (SIP) 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 SIP1,2,4 and LPA(1) transcripts in CHO-K1 cells by RT-PCR. (c) 2005 Elsevier B.V. All rights reserved.

引用

页码：59 / 63

页数：5

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