镰形扇头蜱巨噬细胞移动抑制因子的克隆、表达及酶活性分析  

Clone expression and enzyme activity analysis of macrophage migration inhibitory factor of Rhipicephalus haemaphysaloides

作　　者：王玉俭 郑倩 宋冠华 钟平生 WANG Yujian;ZHENG Qian;SONG Guanhua;ZHONG Pingsheng(School of Life Science,Huizhou University,Huizhou 516007,China)

机构地区：[1]惠州学院生命科学学院,广东惠州516007

出　　处：《黑龙江畜牧兽医》2023年第8期67-71,共5页Heilongjiang Animal Science And veterinary Medicine

基　　金：国家自然科学基金青年基金项目(32102700);广东省基础与应用基础研究基金区域联合基金青年基金项目(2020A1515110318);广东省普通高校青年创新人才类项目(2019KQNCX153);惠州学院自主创新能力提升计划优秀青年培育项目(hzu201803)。

摘　　要：为了探究镰形扇头蜱巨噬细胞移动抑制因子(Rhipicephalus haemaphysaloides macrophage migration inhibitory factor,RhMIF)基因的功能,试验采用RT-PCR方法对RhMIF基因开放阅读框进行扩增,运用大肠杆菌表达体系对RhMIF蛋白进行体外重组表达,随后对纯化后的RhMIF重组蛋白进行互变异构酶活性分析。结果表明:RhMIF基因开放阅读框大小为351 bp,编码116个氨基酸;RhMIF重组蛋白在大肠杆菌上清液中成功表达,大小为17.4 ku;RhMIF重组蛋白具有互变异构酶活性。说明本试验通过大肠杆菌表达体系成功表达了具有酶活性的RhMIF重组蛋白。In order to explore the function of the macrophage migration inhibitory factor gene of Rhipicephalus haemaphysaloides(RhMIF),in this experiment,the open reading frame of the RhMIF gene was amplified by RT-PCR method;the Escherichia coli expression system was used for recombinant expression of RhMIF protein in vitro,and the purified recombinant protein RhMIF was then analyzed for tautomerase activity.The results showed that the open reading frame of the RhMIF gene was 351 bp,encoding 116 amino acids;the RhMIF gene was successfully expressed in the supernatant of E.coli with a size of 17.4 ku;the recombinant protein RhMIF had tautomerase activity.The results suggested that in this experiment,the enzyme-active RhMIF recombinant protein was successfully expressed by the E.coli expression system.

关 键 词：镰形扇头蜱 巨噬细胞移动抑制因子 原核表达 互变异构酶 活性分析 

分 类 号：S852.74[农业科学—基础兽医学]