摘要
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２３０００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．
cDNA fragment encoding mature salmongrowth hormone (sGH) was successfully am-plified by polymerase chain reaction and the re-striction recognition sequences were respec-tively introduced into the 5' end and 3' end ofthe amplified fragment. The recombinant se-cretion vector pOsGH153 containing genescoding for the E. coli ompA signal peptide andmature sGH was constructed and confirmed byrestriction enzyme analysis and Southern blothybridization with the oligonucleotide probe.Upon induction of IPTG, the sGH was ex-pressed in E. coli cells and was secreted intothe E. coli periplasma.
出处
《生物化学与生物物理进展》
SCIE
CAS
CSCD
北大核心
1994年第4期338-341,共4页
Progress In Biochemistry and Biophysics
基金
天津科委自然科学基金
关键词
鲑鱼
生长激素
分泌型
表达质粒
salmon growth hormone, secre-tion vector, periplasma expression, E. coli
