红鳍东方鲀伴侣蛋白编码基因Cctd的克隆与测序

Cloning and sequencing of the chaperonin-encoding Cctd gene from Fugu rubripes rubripes.

作者信息

Yoda T, Morita T, Kawatsu K, Sueki K, Shibata T, Hamano Y

机构信息

Laboratory of Food Microbiology, Osaka Prefectural Institute of Public Health, Japan.

出版信息

Gene. 1995 Dec 12;166(2):249-53. doi: 10.1016/0378-1119(95)00604-4.

DOI:10.1016/0378-1119(95)00604-4

PMID:8543170

Abstract

CCT, a chaperonin containing t-complex polypeptide 1 (TCP-1), is a cytosolic molecular chaperone involved in the folding of proteins. We have isolated the Cctd gene from a Fugu rubripes rubripes (Frr) genomic library using a rat Ccta cDNA as a probe, and cloned its cDNA by reverse transcription-polymerase chain reaction (RT-PCR) using a pair of oligodeoxyribonucleotides corresponding to the 5' and 3' non-coding regions of Frr Cctd. Cctd spans a region of 4.7 kb and consists of at least 13 exons with small introns of about 144 bp on average. The Cctd cDNA sequence revealed a deduced polypeptide of 536 amino acids sharing a high degree of homology with that of the mouse Cctd cDNA (88%). Cctd is present as a single-copy gene, as shown by genomic Southern blot analysis, and can be used for evolutionary and classification analyses of Fugu species.

摘要

CCT是一种包含t-复合体多肽1（TCP-1）的伴侣蛋白，是一种参与蛋白质折叠的胞质分子伴侣。我们以大鼠Ccta cDNA为探针，从红鳍东方鲀（Frr）基因组文库中分离出Cctd基因，并使用一对对应于Frr Cctd 5'和3'非编码区的寡脱氧核糖核苷酸，通过逆转录-聚合酶链反应（RT-PCR）克隆了其cDNA。Cctd跨度为4.7 kb，由至少13个外显子组成，平均含有约144 bp的小内含子。Cctd cDNA序列显示推导的多肽有536个氨基酸，与小鼠Cctd cDNA的多肽具有高度同源性（88%）。基因组Southern印迹分析表明，Cctd作为单拷贝基因存在，可用于红鳍东方鲀物种的进化和分类分析。