从人A431癌细胞中分离出一种进化上保守的表皮生长因子受体cDNA。

Isolation of an evolutionarily conserved epidermal growth factor receptor cDNA from human A431 carcinoma cells.

作者信息

Simmen F A, Gope M L, Schulz T Z, Wright D A, Carpenter G, O'Malley B W

出版信息

Biochem Biophys Res Commun. 1984 Oct 15;124(1):125-32. doi: 10.1016/0006-291x(84)90926-4.

Abstract

Complementary DNA corresponding to total poly(A)+-RNA from the human A431 epidermoid carcinoma cell line was cloned in the phage expression vector lambda gt 11. An epidermal growth factor (EGF) receptor cDNA clone was obtained by screening of the expression library with a rabbit polyclonal antibody (IgG), raised to the purified A431 EGF receptor, in combination with [125I]protein A of S. aureus. The cloned cDNA was able to select, by hybridization, messenger RNA which was translated in Xenopus oocytes and yielded an immunoprecipitable EGF receptor protein of Mr = 160,000. The insert of this cDNA (phEGFR-1), is approximately 880 base pairs in length and encodes the carboxyterminal portion of the EGF receptor protein. Its sequence is evolutionarily conserved among vertebrates as shown by hybridization to unique chromosomal DNA sequences from human, baboon, dog, rat, mouse and frog.

摘要

将来自人A431表皮癌细胞系的总聚腺苷酸加尾RNA（poly(A)+-RNA）对应的互补DNA（cDNA）克隆到噬菌体表达载体λgt 11中。通过用针对纯化的A431表皮生长因子（EGF）受体产生的兔多克隆抗体（IgG）与金黄色葡萄球菌的[125I]蛋白A结合筛选表达文库，获得了EGF受体cDNA克隆。克隆的cDNA能够通过杂交选择在非洲爪蟾卵母细胞中翻译并产生Mr = 160,000的可免疫沉淀的EGF受体蛋白的信使RNA。该cDNA（phEGFR-1）的插入片段长度约为880个碱基对，编码EGF受体蛋白的羧基末端部分。通过与来自人、狒狒、狗、大鼠、小鼠和青蛙的独特染色体DNA序列杂交表明，其序列在脊椎动物中是进化保守的。