Glass C, Kim K H, Fuchs E
J Cell Biol. 1985 Dec;101(6):2366-73. doi: 10.1083/jcb.101.6.2366.
Using mRNA from cultured human mesothelial cells, we constructed bacterial plasmids and lambda phage vectors that contained cDNA sequences specific for the keratins expressed in these cells. A cloned cDNA encoding keratin K7 (55 kD) was identified by positive hybrid selection. Southern Blot analysis indicated that this sequence is represented only once in the human genome, and Northern Blot analysis demonstrated that the gene encoding K7 is expressed in abundance in cultured bronchial and mesothelial cells, but only weakly in cultured epidermal cells and not at all in liver, colon, or exocervical tissue. The predicted amino acid sequence of this keratin has revealed a striking difference between this keratin and the type II keratins expressed in epidermal cells: whereas all of the epidermal type II keratins thus far sequenced have long nonhelical termini rich in glycine and serine, this mesothelial type II keratin has amino and carboxy terminal regions that are unusually short and lack the inexact repeats of glycine and serine residues.
我们使用来自培养的人间皮细胞的mRNA,构建了细菌质粒和λ噬菌体载体,它们包含这些细胞中表达的角蛋白的特异性cDNA序列。通过阳性杂交筛选鉴定出一个编码角蛋白K7(55kD)的克隆cDNA。Southern印迹分析表明该序列在人类基因组中仅出现一次,Northern印迹分析表明编码K7的基因在培养的支气管和间皮细胞中大量表达,但在培养的表皮细胞中表达较弱,在肝脏、结肠或宫颈外组织中则完全不表达。这种角蛋白的预测氨基酸序列显示出该角蛋白与表皮细胞中表达的II型角蛋白之间存在显著差异:迄今为止测序的所有表皮II型角蛋白都有富含甘氨酸和丝氨酸的长非螺旋末端,而这种间皮II型角蛋白的氨基和羧基末端区域异常短,并且缺乏甘氨酸和丝氨酸残基的不精确重复。