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弓形虫GT1虫株GRA15蛋白的表达及其反应原性分析

点击数:21511  时间:2019-03-01 09:02:32  来源: 中国动物检疫      作者: 周芃来等

为了分析弓形虫GT1虫株GRA15GRA15GT1)蛋白的反应原性,通过PCR扩增编码GRA15GT1 52~635氨基酸肽段的基因片段,构建pGEX-6P-1-GRA15GT1载体,转化BL21菌诱导表达;通过SDS-PAGEWestern blot方法进行表达验证及反应原性分析。结果显示:SDS-PAGE及以GST标签抗体为一抗进行Western blot,均有目的条带,比理论值稍大;以猪弓形虫阳性血清为一抗的Western blot条带与GST抗体孵育后的条带大小一致。上述结果表明,GRA15GT1蛋白具有较好的反应原性,这为下一步分段表达GRA15GT1蛋白,研究其在弓形虫血清学分型中的应用奠定了基础。


Expression and Immunoreactivity Exploration of GRA15Protein

of Toxoplasma gondii GT1 Strain

In order to analyze the immunoreactivity of GRA15protein of Toxoplasma gondii GT1 strainGRA15GT1),the gene encoding a 584-residue peptidefrom aa52 to aa635was amplified by PCRthen the pGEX-6P-1-GRA15GT1 vector wasconstructed and transformed into BL21 strain of E. coli. After inducibleexpressionthe purified GRA15 protein were obtainedand verified by SDS-PAGE and Western blot. Results showed thatthe target strip was slightly larger thanits estimated size when the GST-tag mouse monoclonal antibody was used asprimary antibodiesas well as the swine anti-T. gondii positive serum. The resultsindicated GRA15GT1 protein had good immunoreactivitywhich would lay a foundation for the next stepof GRA15GT1 protein expression and its application in serological typing ofToxoplasma gondii.

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