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人生长激素:互补DNA的克隆及在细菌中的表达

Human growth hormone: complementary DNA cloning and expression in bacteria.

作者信息

Martial J A, Hallewell R A, Baxter J D, Goodman H M

出版信息

Science. 1979 Aug 10;205(4406):602-7. doi: 10.1126/science.377496.

Abstract

The nucleotide sequence of a DNA complementary to human growth hormone messenger RNA was cloned; it contains 29 nucleotides in its 5' untranslated region, the 651 nucleotides coding for the prehormone, and the entire 3' untranslated region (108 nucleotides). The data reported predict the previously unknown sequence of the signal peptide of human growth hormone and, by comparison with the previously determined sequences of rat growth hormone and human chorionic somatomammotropin, strengthens the hypothesis that these genes evolved by gene duplication from a common ancestral sequence. The human growth hormone gene sequences have been linked in phase to a fragment of the trp D gene of Escherichia coli in a plasmid vehicle, and a fusion protein is synthesized at high level (approximately 3 percent of bacterial protein) under the control of the regulatory region of the trp operon. This fusion protein (70 percent of whose amino acids are coded for by the human growth hormone gene) reacts specifically with antibodies to human growth hormone and is stable in E. coli.

摘要

克隆了与人生长激素信使核糖核酸互补的一段DNA的核苷酸序列;它在其5'非翻译区含有29个核苷酸,编码前激素的651个核苷酸,以及整个3'非翻译区(108个核苷酸)。所报道的数据预测了人生长激素信号肽以前未知的序列,并且通过与大鼠生长激素和人绒毛膜促生长催乳素先前确定的序列相比较,强化了这样一种假说,即这些基因是通过基因复制从一个共同的祖先序列进化而来的。人生长激素基因序列已在质粒载体中与大肠杆菌色氨酸D基因的一个片段同框连接,并且在色氨酸操纵子调节区的控制下高水平合成一种融合蛋白(约占细菌蛋白的3%)。这种融合蛋白(其氨基酸的70%由人生长激素基因编码)与人生长激素抗体发生特异性反应,并且在大肠杆菌中是稳定的。

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