Wenzel K, Manthey D, Willecke K, Grzeschik K H, Traub O
Abt. Molekulargenetik, Universität Bonn, Germany.
Biochem Biophys Res Commun. 1998 Jul 30;248(3):910-5. doi: 10.1006/bbrc.1998.9070.
We have isolated and characterized a human genomic clone containing the complete coding region of connexin31 (Cx31). Similar to rodent Cx31, the coding region of human Cx31 is completely contained within the second exon and consists of 810 nucleotides. The deduced human Cx31 polypeptide consists of 270 amino acids with a predicted molecular mass of 30.818 kDa. Its sequence is most similar to mouse Cx31 (82.6% identical amino acids) and rat (83.0% identical amino acids), but shows considerably fewer potential sites of phosphorylation. After Northern blot hybridization, two Cx31 transcripts of 2.2 and 1.8 kb were detected in total RNA of the human keratinocyte cell line HaCaT and two transcripts of 2.2 and 1.9 kb in total RNA of E6/E7 transfected human keratinocytes (HEK cells). Using affinity-purified rabbit antibodies to mouse Cx31, immunofluorescence analysis demonstrated relatively weak expression of human Cx31 in HaCaT and HEK cells. The Cx31 gene exists as a single copy gene in the human genome and was mapped to the chromosomal region 1p34-p36 by analyzing human-mouse somatic cell hybrids.
我们分离并鉴定了一个包含连接蛋白31（Cx31）完整编码区的人类基因组克隆。与啮齿动物的Cx31相似，人类Cx31的编码区完全包含在第二个外显子内，由810个核苷酸组成。推导的人类Cx31多肽由270个氨基酸组成，预测分子量为30.818 kDa。其序列与小鼠Cx31（82.6%氨基酸相同）和大鼠Cx31（83.0%氨基酸相同）最相似，但磷酸化潜在位点明显较少。经过Northern印迹杂交，在人类角质形成细胞系HaCaT的总RNA中检测到2.2 kb和1.8 kb的两种Cx31转录本，在E6/E7转染的人类角质形成细胞（HEK细胞）的总RNA中检测到2.2 kb和1.9 kb的两种转录本。使用针对小鼠Cx31的亲和纯化兔抗体，免疫荧光分析显示人类Cx31在HaCaT和HEK细胞中的表达相对较弱。Cx31基因在人类基因组中作为单拷贝基因存在，并通过分析人鼠体细胞杂种被定位到染色体区域1p34 - p36。
