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当前位置: 首页 > 文章 > 双芽巴贝斯虫RAP-1C基因的克隆表达及重组蛋白反应原性的研究 中国兽医科学 2014 (8) 823-828
Position: Home > Articles > Cloning and expression of RAP-1C gene fromBabesia bigemina and reactivity of the expressed recombinant protein Chinese Veterinary Science 2014 (8) 823-828

双芽巴贝斯虫RAP-1C基因的克隆表达及重组蛋白反应原性的研究

作  者：
袁利芹;关贵全;刘军龙;赵帅阳;杨聪山
单  位：
中国农业科学院兰州兽医研究所家畜疫病病原生物学国家重点实验室甘肃省动物寄生虫病重点实验室;石河子大学
关键词：
双芽巴贝斯虫;RAP-1C基因;原核表达;免疫原性
摘  要：
拟通过对双芽巴贝斯虫棒状体相关蛋白1(rhoptry-associated protein-1,RAP-1)羧基端进行研究,以期了解该蛋白的一些免疫学特性。利用生物信息学软件,对RAP-1进行抗原表位预测,并与其他重要的巴贝斯虫和泰勒虫相应基因序列进行比对,以筛选出抗原性和特异性均良好的基因片段;针对筛选出的基因片段设计引物,对该基因进行克隆和原核表达,最后利用Western-blot对重组蛋白的反应原性和特异性进行分析。结果显示,融合目的蛋白含His标签,分子质量为26ku左右,该蛋白只与双芽巴贝斯虫阳性血清发生特异性反应,与其他主要巴贝斯虫、泰勒虫的阳性血清无交叉反应。结果表明,表达的PAP-1蛋白可作为ELISA诊断方法的候选抗原,为建立一种双芽巴贝斯虫病特异性的免疫学诊断方法奠定了基础。
译  名：
Cloning and expression of RAP-1C gene fromBabesia bigemina and reactivity of the expressed recombinant protein
作  者：
YUAN Li-qin;GUAN Gui-quan;LIU Jun-long;ZHAO Shuai-yang;YANG Cong-shan;LU Hai-yan;YIN Hong;MENG Qing-ling;LUO Jian-xun;College of Animal Science and Technology,Shihezi University;State Key Laboratory of Veterinary Etiological/Biology Key Laboratory of Veterinary Parasitology of Gansu Province/Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences;
关键词：
Babesia bigemina;;C-terminal of rhoptry-associated protein-1(RAP-1C)gene;;prokaryotic expression;;immunogenicity
摘  要：
The purpose of this research is to understand the potential immunological traits of the C-terminal fragment of rhoptry-associated protein-1(RAP-1)of Babesia bigemina.The C-terminal fragment of RAP-1of B.bigemina with high antigenicity and specificity was screened by using bioinformatics analysis and sequence alignment with these of other important Babesia and Theileria species.The RAP-1Cgene was cloned and expressed in prokaryotic expression system.The reactivity and specificity of the protein were evaluated by Western-blot.In result,the fusion protein with His was 26ku in size.The recombinant protein had strong reactions with the positive sera fromB.bigeminainfected cattle but no cross-reactions with positive sera against other Babesiaand Theileriaspecies.In conclusion,the recombinant protein RAP-1Cwill be used as a candidate of diagnostic antigen to develop an ELISA technique,which lays the foundation for the development of immuno-diagnostic methods for specifically detecting B. bigemina infection.

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