长角血蜱巨噬细胞移动抑制因子的克隆表达及酶活分析  

Cloning,Expression and Enzyme Activity Analysis of Macrophage Migration Inhibitor from Haemaphysalis longicornis

作　　者：王玉俭 代绍密 李晓燕 黄冰冰 宋冠华 钟平生 WANG Yujian;DAI Shaomi;LI Xiaoyan;HUANG Bingbing;SONG Guanhua;ZHONG Pingsheng(School of Life Science,Huizhou University,Huizhou 516007,Guangdong,China)

机构地区：[1]惠州学院生命科学学院,广东惠州516007

出　　处：《惠州学院学报》2022年第6期34-38,共5页Journal of Huizhou University

基　　金：国家自然科学基金项目(32102700);广东省基础与应用基础研究基金区域联合基金项目(2020A1515110318)。

摘　　要：为探究长角血蜱(Haemaphysalis longicornis)巨噬细胞移动抑制因子(Macrophage migration inhibitory factor,MIF)基因(HlMIF)的功能,采用RT-PCR方法对HlMIF基因进行扩增,运用大肠杆菌表达体系对HlMIF基因进行体外重组表达,随后对纯化后的重组HlMIF蛋白进行互变异构梅活性分析。结果表明:HlMIF基因开放阅读框为351 bp,编码116个氨基酸,预测成熟蛋白的分子量为12.5 ku;HlMIF基因在大肠杆菌上清中成功表达,大小为17.4ku;重组HlMIF蛋白具有互变异构酶活性;利用大肠杆菌成功表达具有酶活力的重组HlMIF蛋白,为进一步明确该蛋白的生物学功能提供条件。In order to explore the function of the macrophage migration inhibitor gene of Haemaphysalis longicornis,the HlMIF gene was amplified by RT-PCR,and the HlMIF gene was recombinantly expressed in vitro by using the Escherichia coli expression system,the purified recombinant HlMIF protein was then subjected to tautomeric activity analysis. The results showed that the open reading frame of the HlMIF gene was 351 bp,encoding 116 amino acids,and the predicted molecular weight of the mature protein was 12.5 ku;the HlMIF gene was successfully expressed in the supernatant of E. coli with a size of 17.4 ku;the recombinant HlMIF protein has tautomerase activity. The recombinant HlMIF protein with enzymatic activity was successfully expressed in E. coli,which provided conditions for further clarifying the biological function of the protein.

关 键 词：长角血蜱 巨噬细胞移动抑制因子 原核表达 互变异构酶活性 

分 类 号：S852.74[农业科学—基础兽医学]