Cloning and sequence analysis of rat cystic fibrosis transmembrane conductance regulator.
Cloning and sequence analysis of rat cystic fibrosis transmembrane conductance regulator.
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大鼠囊性纤维化跨膜电导调节因子的克隆及序列分析。
DOI：
10.1152/ajplung.1992.262.6.l779
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发表时间：
1992
期刊：
影响因子：
--
通讯作者：
Shull,GE
中科院分区：
文献类型：
--
作者：
Fiedler,MA;Nemecz,ZK;Shull,GE
已分离出编码大鼠囊性纤维化跨膜电导调节因子 (CFTR) 的互补 DNA (cDNA)，并测定了大鼠 CFTR mRNA 的组织分布。 Northern 印迹分析显示，6.3 kb CFTR mRNA 在结肠中表达水平最高，在子宫、肺、胃和小肠中也有表达。 7.5-kb mRNA在骨骼肌中表达，并且在睾丸中表达7.5-kb mRNA和6.0-kb mRNA。从大鼠结肠文库中分离出 5 个 cDNA，最长的对应于通过聚 A 尾的密码子 684。还分离了对应于密码子213至245、372至574和656至886的其他三个克隆。利用从大鼠结肠mRNA制备的cDNA的聚合酶链式反应扩增来克隆cDNA的其余部分。大鼠CFTR的预测氨基酸序列与人CFTR有79%相同，其中R结构域有73%同一性，核苷酸结合折叠1和2分别有81%和83%同一性。在人类 CFTR 中发现的 38 个潜在磷酸化位点中，有 32 个在大鼠 CFTR 中也存在。
A complementary DNA (cDNA) encoding the rat cystic fibrosis transmembrane conductance regulator (CFTR) has been isolated and the tissue distribution of the rat CFTR mRNA has been determined. Northern blot analysis revealed that the highest levels of the 6.3 kilobase (kb) CFTR mRNA were expressed in the colon, with expression also noted in uterus, lung, stomach, and small intestine. A 7.5-kb mRNA was expressed in skeletal muscle, and in testes both the 7.5-kb mRNA and a 6.0-kb mRNA were expressed. Five cDNAs were isolated from a rat colon library, the longest corresponding to codons 684 through the poly A tail. Three other clones, corresponding to codons 213 through 245, 372 through 574, and 656 through 886 were also isolated. Polymerase chain reaction amplification of cDNA prepared from rat colon mRNA was utilized to clone the remainder of the cDNA. The predicted amino acid sequence of the rat CFTR is 79% identical to the human CFTR, with 73% identity noted in the R domain, and 81 and 83% identities noted in nucleotide binding folds 1 and 2, respectively. Thirty-two of the 38 potential phosphorylation sites identified in the human CFTR were also present in the rat CFTR.