Isolation and Expression Profile of the Ca2+-Activated Chloride Channel-like Membrane Protein 6 Gene in Xenopus laevis

Title
Isolation and Expression Profile of the Ca2+-Activated Chloride Channel-like Membrane Protein 6 Gene in Xenopus laevis
Authors
이라미류래형정성원오수진Hue Huang한진수이치호이창준Lily Yeh Jan정상민
Keywords
CMP6; calcium-activated chloride channel; Xenopus laevis; gene cloning; tissue distribution; expression profile; real time-PCR
Issue Date
2011-06
Publisher
Laboratory Animal Research
Citation
VOL 27, NO 2, 109-116
Abstract
To clone the first anion channel from Xenopus laevis (X. laevis), we isolated a calcium-activated chloride channel (CLCA)-like membrane protein 6 gene (CMP6) in X. laevis. As a first step in gene isolation, an expressed sequence tags database was screened to find the partial cDNA fragment. A putative partial cDNA sequence was obtained by comparison with rat CLCAs identified in our laboratory. First stranded cDNA was synthesized by reverse transcription polymerase-chain reaction (RT-PCR) using a specific primer designed for the target cDNA. Repeating the 5' and 3' rapid amplification of cDNA ends, fulllength cDNA was constructed from the cDNA pool. The full-length CMP6 cDNA completed via 5'- and 3'- RACE was 2,940 bp long and had an open reading frame (ORF) of 940 amino acids. The predicted 940 polypeptides have four major transmembrane domains and showed about 50% identity with that of rat brain CLCAs in our previously published data. Semi-quantification analysis revealed that CMP6 was most abundantly expressed in small intestine, colon and liver. However, all tissues except small intestine, colon and liver had undetectable levels. This result became more credible after we did real-time PCR quantification for the target gene. In view of all CLCA studies focused on human or murine channels, this finding suggests a hypothetical protein as an ion channel, an X. laevis CLCA.
URI
http://pubs.kist.re.kr/handle/201004/40161
ISSN
1738-6055
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KIST Publication > Article
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