鲑鱼生长激素基因分泌型表达质粒的构建

生长激素(GH)是动物垂体前叶分泌的一种多肽类激素.应用分子重组及PCR等技术,构建了一种鲑鱼生长激素基因分泌型表达质粒pOsGH153,使编码鲑鱼生长激素成熟肽的序列克隆在大肠杆菌分泌型表达载体PIN-Ⅲ-ompA内,直接位于编码大肠杆菌外膜蛋白A信号肽序列的下游,在Lpp-Lac杂合启动子控制下,经IPTG诱导,分子量约23 000的鲑鱼生长激素在大肠杆菌中获得高效表达,该产物具有天然鲑鱼生长激素的免疫活性,直接分泌到细胞周质,而信号肽被自动剪除.

Expression and Secretion of Salmon Growth Hormone From an Escherichia coli Secretion Vector

A cDNA fragment encoding mature salmon growth hormone (sGH) was successfully amplified by polymerase chain reaction and the restriction recognition sequences were respectively introduced into the 5' end and 3' end of the amplified fragment. The recombinant secretion vector pOsGH153 containing genes coding for the E.coli ompA signal peptide and mature sGH was constructed and confirmed by restriction enzyme analysis and Southern blot hybridization with the oligonucleotide probe. Upon induction of IPTG, the sGH was expressed in E.coli cells and was secreted into the E.coli periplasma.