Fiddes J C, Seeburg P H, DeNoto F M, Hallewell R A, Baxter J D, Goodman H M
Proc Natl Acad Sci U S A. 1979 Sep;76(9):4294-8. doi: 10.1073/pnas.76.9.4294.
A 2.6-kilobase (kb) EcoRI restriction endonuclease fragment containing human growth hormone (hGH; somatotropin) gene sequences and a 2.8-kb EcoRI fragment containing human chorionic somatomammotropin (hCS; choriomammotropin) gene sequences have been identified by hybridization to cloned cDNA. Human DNA was cleaved with EcoRI and fractionated by preparative agarose gel electrophoresis; DNA in the size range 2--3 kb was ligated to lambda gt WES.lambda B DNA and viable recombinant bacteriophage were recovered by in vitro packaging. After infection of Escherichia coli and screening of phage plaques, single isolates of hGH and hCS gene sequences were obtained. Restriction endonuclease mapping showed that the hGH gene contains three intervening sequences interrupting the coding sequence. Partial DNA sequence analysis of the hGH gene, obtained by the chain termination method, confirmed the location of the intervening sequences and the identity of the fragment.
通过与克隆的互补DNA(cDNA)杂交,已鉴定出一个含有人生长激素(hGH;促生长素)基因序列的2.6千碱基(kb)的EcoRI限制性内切酶片段和一个含有人类绒毛膜生长催乳素(hCS;绒毛膜促乳腺素)基因序列的2.8 kb的EcoRI片段。用人的DNA与EcoRI进行酶切,并通过制备性琼脂糖凝胶电泳进行分级分离;将大小在2 - 3 kb范围内的DNA连接到λgt WES.λB DNA上,并通过体外包装回收有活力的重组噬菌体。在感染大肠杆菌并筛选噬菌体噬菌斑后,获得了hGH和hCS基因序列的单克隆分离株。限制性内切酶图谱分析表明,hGH基因包含三个中断编码序列的间隔序列。通过链终止法获得的hGH基因的部分DNA序列分析证实了间隔序列的位置以及该片段的一致性。