| 牛巴贝斯虫新疆株MSA-2c基因的克隆表达及其重组蛋白免疫原性 |
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| 引用本文： | 简子健,袁江玲,马素贞,黄家雨,沈炯玉.牛巴贝斯虫新疆株MSA-2c基因的克隆表达及其重组蛋白免疫原性[J].中国兽医学报,2009,29(10). |
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| 作者姓名： | 简子健 袁江玲 马素贞 黄家雨 沈炯玉 |
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| 作者单位： | 新疆农业大学,动物医学院,新疆,乌鲁木齐,830052 |
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| 摘 要： | 本试验将新疆株牛巴贝斯虫的MSA-2c基因克隆,并构建重组质粒pGEX-4T-2/MSA-2c.利用大肠杆菌原核表达系统进行外源蛋白的表达,并成功诱导出GST-MSA-2c融合蛋白.通过优化诱导条件,得到了较高的可溶性表达;利用亲和层析技术纯化后的重组蛋白皮下免疫试验小鼠.间接ELISA检测发现,免疫接种56 d后,抗重组蛋白的抗体效价达到1∶220 000以上.用获得的抗血清进行Western-blot试验,可获得清晰的免疫反应条带.结果表明,本试验所表达的MSA-2c蛋白与文献报道的目的蛋白相符,并具有免疫原性.同时本试验也为建立以MSA-2c重组蛋白作为抗原的血清学诊断体系奠定了基础.
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| 关 键 词： | 牛巴贝斯虫 MSA-2c 重组表达 免疫原性 |
Cloning and expression of merozoite surface antigen 2 locus(MSA-2c) of Babesia bovis isolated in Xinjiang and antigenicity of its recombinant protein |
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| JIAN Zi-jian,YUAN Jiang-ling,MA Su-zhen,HUANG Jia-yu,SHEN Jiong-yu.Cloning and expression of merozoite surface antigen 2 locus(MSA-2c) of Babesia bovis isolated in Xinjiang and antigenicity of its recombinant protein[J].Chinese Journal of Veterinary Science,2009,29(10). |
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| Authors: | JIAN Zi-jian YUAN Jiang-ling MA Su-zhen HUANG Jia-yu SHEN Jiong-yu |
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| Abstract: | The merozoite surface antigen 2 locus gene(MSA-2c) of Babesia bovis found in Xinjiang was cloned and ligated into pGEX-4T-2 to construct a recombinant plasmid,pGEX-4T-2/MSA-2c.A fusion protein,GST-MSA-2c,was successfully expressed through prokaryotic expression system in E.coli.The high quantity of soluble expression was obtained by improving the induction conditions.The recombinant protein purified by affinity chromatography was inoculated into mice subcutaneously.The antibody titers of the recombinant protein in murine sera were above 1∶220 000 at 56 days postinoculation by indirect ELISA.The antisera were then examined by Western-blot and a clear reaction strip was found.The results showed that of the expressed MSA-2c protein was identified with interest protein reported in records,and was highly immunogenic(antigenic).It laysa foundation of construction serologically diagnostic system using MSA-2c recombinant protein as a diagnostic antigen. |
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| Keywords: | Babesia bovis MSA-2c recombinant expression immunogenicity |
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