| 弓形虫复合基因SAG1-BAG1的构建、表达及重组抗原的免疫反应性分析 |
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| 引用本文： | 秘文婷,姚硕,钟志刚,杨培梁,吴焜,陈晓光. 弓形虫复合基因SAG1-BAG1的构建、表达及重组抗原的免疫反应性分析[J]. 热带医学杂志, 2012, 12(5): 620-622 |
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| 作者姓名： | 秘文婷 姚硕 钟志刚 杨培梁 吴焜 陈晓光 |
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| 作者单位： | 南方医科大学公共卫生与热带医学学院病原生物学系,广东广州,510515 |
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| 基金项目： | 广东省自然科学基金(8451051501000253);广东省科技计划项目(2008A060202021);广东省大学生创新实验项目(1212110025、1212110027);南方医科大学学生课外科研项目(2009kw092、2009kw093) |
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| 摘 要： | 目的构建并表达弓形虫复合基因SAG1-BAG1及分析重组抗原的免疫反应性。方法利用PCR扩增弓形虫SAG1基因,利用RT-PCR扩增弓形虫BAG1基因,通过酶切连接分别将SAG1、BAG1基因克隆到pET28a表达载体中构建复合基因SAG1-BAG1。将质粒pET28a-SAG1-BAG1转入大肠杆菌BL21(DE3)中进行诱导表达,Western-blot分析该重组蛋白的免疫反应性。结果复合基因SAG1-BAG1全长约1 407 bp,重组蛋白相对分子质量约为50 000,表达量约占菌体总蛋白的10%,Western-blot分析显示重组蛋白具有良好的免疫反应性。结论成功构建并表达弓形虫复合基因SAG1-BAG1。
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| 关 键 词： | 弓形虫 复合基因 SAG1 BAG1 蛋白表达 |
Construction,expression and identification of SAG1-BAG1 complex gene from Toxoplasma gondii |
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| MI Wen-ting , YAO Shuo , ZHONG Zhi-gang , YANG Pei-liang , WU Kun , CHEN Xiao-guang. Construction,expression and identification of SAG1-BAG1 complex gene from Toxoplasma gondii[J]. Journal Of Tropical Medicine, 2012, 12(5): 620-622 |
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| Authors: | MI Wen-ting YAO Shuo ZHONG Zhi-gang YANG Pei-liang WU Kun CHEN Xiao-guang |
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| Affiliation: | (Department of Etio-Biology,School of Public Health and Tropical Medicine,Southern Medical University,Guangdong,Guangzhou 510515,China) |
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| Abstract: | Objective To construct,express and identify SAG1-BAG1 complex gene from Toxoplasma gondii.Methods The SAG1 gene and BAG1 gene were amplified by PCR and RT-PCR,respectively.And the two genes were cloned into the same pET28a vector to construct SAG1-BAG1 complex gene.After induced by IPTG,the recombinant protein was identified by Western-blot.Results The SAG1-BAG1 complex gene was about 1 407 bp.The recombinant protein was expressed successfully,and was about 50 kDa in size.Western-blot showed that the protein could be identified specifically by Toxoplasma gondii infected serum.Conclusion The complex gene SAG1-BAG1 was constructed successfully. |
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| Keywords: | Toxoplasma gondii complex gene SAG1 BAG1 protein expression |
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