弓形虫MIC10基因的克隆与表达  

Cloning and expression of the Toxoplasma gondii MIC10 gene

作　　者：张煊承 李航[1] 谢素珠 赵少伟 王浩[1] 张爽[1] 贾立军 ZHANG Xuancheng;LI Hang;XIE Suzhu;ZHAO Shaowei;WANG Hao;ZHANG Shuang;JIA Lijun(MOE Engineering Research Center of Innovative Science and Technology for Beef Cattle in the North-East Cold Region/Yanbian University,Yanji 133002,China)

机构地区：[1]东北寒区肉牛科技创新教育部工程研究中心,延边大学,吉林延吉133002

出　　处：《畜牧与兽医》2020年第8期75-78,共4页Animal Husbandry & Veterinary Medicine

基　　金：国家自然科学基金(31760729);高等学校学科创新引智111计划资助(D20034);吉林省中青年科技创新领军人才及科研创新团队项目(20200301034RQ);吉林省人才开发资金集中资助人才项目(吉财社指[2019]874号)。

摘　　要：弓形虫MIC10基因在速殖子时期与缓殖子时期都存在不同程度的表达,但国内对该基因的研究甚少。为了解弓形虫MIC10基因的功能,构建其原核表达载体,并进行原核表达。本试验以提取的弓形虫DNA为模板,设计引物扩增MIC10基因片段,连接克隆载体pMD-18T和表达载体PGEX-4T-1,PCR和双酶切鉴定后进行原核表达。结果表明,经PCR扩增获得597 bp的MIC10基因片段,构建pMD18-T-MIC10克隆质粒和PGEX-4T-MIC10原核表达载体,经PCR鉴定和双酶切鉴定正确,并在大肠杆菌中成功表达,表达蛋白的分子量约为49 kD。本试验为弓形虫MIC10基因功能的后续研究奠定了基础。The MIC10 gene of Toxoplasma gondii is expressed in different degrees in tachyzoites and bradyzoites,but little research has been done on this gene in China.In order to understand the function of MIC10 of Toxoplasma gondii,the prokaryotic expression vector of the gene was constructed in this experiment and prokaryotic expression was performed.The DNA extracted from Toxoplasma gondii was used as template,and the primers were designed to amplify the MIC10 fragment.The cloning vector and the expression vector were connected.After identification by PCR and double digestion,the prokaryotic expression was carried out.The results showed that a 597 bp MIC10 fragment was amplified by PCR,and the pMD18-T-MIC10 clone plasmid and the prokaryotic expression vector of PGEX-4 T-MIC10 were constructed.The expression was confirmed by PCR and double digestion.The molecular weight of the expressed protein was about 49 kD.This experiment laid a foundation for the follow-up study of the function of the MIC10 gene of Toxoplasma gondii.

关 键 词：弓形虫 MIC10基因 克隆 原核表达 

分 类 号：S852.7[农业科学—基础兽医学]