鸡生长激素的原核表达、纯化及活性研究  

Prokaryotic Expression,Purification and Activity Study of Chicken Growth Hormone

作　　者：何赛[1,2] 周欢庆[1] 陈昱希[1] 李洁[1] 郁建锋[1] 顾志良[1] 

机构地区：[1]常熟理工学院,江苏常熟215500 [2]中国矿业大学化工学院,江苏徐州221000

出　　处：《中国家禽》2016年第5期5-9,共5页China Poultry

基　　金：国家自然科学基金项目(31072025);苏州市科技计划项目(SYN201416)

摘　　要：鸡生长激素(cGH)是由脑垂体前叶合成并分泌的一种重要的多肽类激素,对于鸡的生长发育具有至关重要的调控作用。为实现cGH的原核表达,将其序列插入原核表达载体pET-30a(+)中,构建重组表达质粒pET-30a-cGH,并转化大肠杆菌BL21(DE3),挑选阳性菌落液体培养IPTG诱导表达。SDS-PAGE检测显示,诱导产物在25 ku处含有与预期大小一致的蛋白条带;通过镍柱纯化,得到了较高纯度的cGH重组蛋白;用纯化得到的cGH重组蛋白处理鸡肝癌LMH细胞,检测其IGF-Ⅰ基因mRNA水平上调,证明所得到cGH蛋白具有生物学活性。研究结果为进一步研究cGH的结构、功能和生产应用奠定了基础。As an important polypeptide hormone,chicken growth hormone（cGH）is synthesized and excreted by anterior pituitary,which plays a vital role in adjusting the growth and development of body. To achieve recombinant cGH from prokaryotes,cGH gene was cloned into a prokaryotic expression vector,pET-30 a,to construct pET-30a-cGH. The recombinant plasmid was transformed to E.coli BL21（DE3）,and then the positive bacterial strain was induced with IPTG. The results of SDS-PAGE indicated that the induced products contained a 25 ku protein as expected,and a highly pure recombinant cGH protein was purified by Ni-sepharose. The level of IGF-Ⅰ mRNA in LMH cells increased after treated with recombinant cGH,it certified that the recombinant cGH protein had biological activity. This would lay the groundwork for the further study of structure,function and application in production of cGH.

关 键 词：鸡 生长激素 原核表达 蛋白纯化 活性研究 

分 类 号：S831.2[农业科学—畜牧学]