| 马泰勒虫新疆株EMA2基因的克隆及原核表达 |
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| 引用本文： | 张杨,刘世芳,海尼木古力·艾合买提,王盼举,宋瑞其,巴音查汗. 马泰勒虫新疆株EMA2基因的克隆及原核表达[J]. 中国动物传染病学报, 2019, 0(4): 88-91 |
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| 作者姓名： | 张杨 刘世芳 海尼木古力·艾合买提 王盼举 宋瑞其 巴音查汗 |
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| 作者单位： | 新疆农业大学动物医学学院;托克逊县夏乡畜牧业服务中心 |
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| 基金项目： | 新疆维吾尔自治区自然科学基因(2016D01B027) |
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| 摘 要： | 裂殖子表面抗原蛋白家族因其高抗原保守性常作为马泰勒虫ELISA检测方法的基质。本研究根据马泰勒虫EMA2基因序列合成引物,以马泰勒虫新疆株DNA为模板,扩增获得目的基因,将其连接至pEASY-E1载体构建重组质粒pEASY-E-TE,并转化至大肠杆菌BL21中,IPTG诱导表达重组EMA2蛋白并鉴定。结果显示:PCR扩增获得了约819bp特异性DNA片段,构建了pEASY-E-TE重组质粒,成功诱导表达了35kDa的可溶性目的蛋白;经SDS-PAGE、Western blot鉴定,重组蛋白rEMA2具有良好的抗原性。本试验克隆表达的重组蛋白rEMA2可为ELISA方法的建立及后期研究提供靶基因材料。
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| 关 键 词： | 马泰勒虫 新疆虫株 EMA2基因 原核表达 |
CLONING AND PROKARYOTIC EXPRESSION OF MEROZOITE ANTIGEN 2 GENE OF THEILERIA EQUI XINJIANG STRAIN |
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| Affiliation: | (College of Animal Medicine,Xinjiang Agriculture University,Urumqi 830052,China;Animal Husbandary Service Center of Toksun County,Turpan 838100,China) |
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| Abstract: | Merozoite antigen family is highly conserved and commonly used in ELISA diagnostic testing for Theileria equi.In this study,primers were designed to amplify the EMA2 gene of T.equi Xinjiang strain and to construct the recombinant plasmid pEASY-E-TE.Then plasmid pEASY-E-TE was transformed into E.coli BL21 competent cells for gene expression.The result showed that the PCR product was 819 bp in length and expressed 35 kDa protein was identified in SDS-PAGE and Western blot.rEMA2 from prokaryotic expression laid a foundation for development of ELISA diagnostic testing and research of its function. |
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| Keywords: | Theileria equi Xinjiang strain EMA2 gene prokaryotic expression |
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