CLONING OF A PUTATIVE HUMAN VOLTAGE-GATED CHLORIDE CHANNEL (CIC-2) CDNA WIDELY EXPRESSED IN HUMAN TISSUES
CLONING OF A PUTATIVE HUMAN VOLTAGE-GATED CHLORIDE CHANNEL (CIC-2) CDNA WIDELY EXPRESSED IN HUMAN TISSUES
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DOI：
10.1093/hmg/4.3.407
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发表时间：
1995-03-01
影响因子：
3.5
通讯作者：
CUTTING, GR
中科院分区：
文献类型：
--
作者：
CID, LP;MONTROSERAFIZADEH, C;CUTTING, GR
我们从人上皮细胞系T84中克隆了一个cDNA，其预测的氨基酸序列与大鼠ClC-2的一致性为93.9%。通过体细胞杂交和聚合酶链式反应进行作图，将该 cDNA 对应的基因定位于染色体 3q26-qter。 RNA 引物延伸评估的主要转录起始位点位于推定翻译起始密码子上游 100 nt 处。对 5' 侧翼序列的分析显示其 GC 含量较高，并且缺乏常见的转录元件，例如 TATA 和 CCAAT 盒。 Northern 印迹分析表明广泛的器官分布，包括囊性纤维化 (CF) 受影响的组织以及来自 CF 患者的气道上皮细胞系中的表达。与大鼠 ClC-2 的高度序列相似性和相似的组织分布表明该 cDNA 编码人 ClC-2 电压门控氯离子通道。由于这种氯离子通道存在于上皮组织中，因此可以通过操作来规避 CF 中观察到的氯离子分泌缺陷。
We have cloned a cDNA from the human epithelial cell line T84 whose predicted amino acid sequence shows 93.9% identity with rat ClC-2. Mapping by somatic cell hybrids and polymerase chain reaction localizes the gene corresponding to this cDNA to chromosome 3q26-qter. The major transcription start site assessed by RNA primer extension is 100 nt upstream of the putative translation initiation codon. Analysis of the 5' flanking sequence revealed a high GC content and lack of common transcriptional elements such as TATA and CCAAT boxes. Northern blot analysis indicated wide organ distribution including tissues affected in cystic fibrosis (CF) and expression in an airway epithelial cell line derived from a CF patient. The high degree of sequence similarity and similar tissue distribution to rat ClC-2 suggests that this cDNA encodes the human ClC-2 voltage-gated chloride channel. Since this chloride channel is present in epithelial tissues it may be amenable to manipulation to circumvent the chloride secretion defect observed in CF.