이라미 류래형 정성원 오수진 Hue Huang 한진수 이치호 이창준 Lily Yeh Jan 정상민 2024-01-20T16:34:30Z 2024-01-20T16:34:30Z 2022-01-10 2011-07 1738-6055 https://pubs.kist.re.kr/handle/201004/130190 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 940polypeptides 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. English 한국실험동물학회 Isolation and Expression Profile of the Ca2+-Activated Chloride Channel-like Membrane Protein 6 Gene in Xenopus laevis Article 2 Laboratory Animal Research, v.27, no.2, pp.109 - 116 Laboratory Animal Research 27 2 109 116 N kci ART001566773 CMP6 calcium-activated chloride channel Xenopus laevis gene cloning tissue distribution expression profile real time-PCR