| 刚地弓形虫GJS株微线体蛋白基因MIC3的克隆与原核表达 |
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| 引用本文： | 苟惠天,王艳华,李文卉,李航,付宝权,张德林.刚地弓形虫GJS株微线体蛋白基因MIC3的克隆与原核表达[J].甘肃农业大学学报,2008,43(5). |
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| 作者姓名： | 苟惠天 王艳华 李文卉 李航 付宝权 张德林 |
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| 作者单位： | 1. 甘肃农业大学动物医学院,甘肃,兰州,700070;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生点实验室,甘肃省动物寄生虫病重点实验室,甘肃 兰州 730046
2. 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生点实验室,甘肃省动物寄生虫病重点实验室,甘肃 兰州 730046 |
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| 基金项目： | 甘肃省生大专项科技项目 |
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| 摘 要： | 根据Genbank中编码MIC3的已知基因序列设计并合成一对引物,应用PCR技术对刚地弓形虫GJS株微线体蛋白基因MIC3进行克隆、表达及鉴定.结果表明:克隆的MIC3基因的开放阅读框与Genbank收录的MIC3基因在核菌酸和氨基酸水平上高度一致,说明弓形虫MIC3蛋白的高度保守性;构建的原核细胞表达质粒PET-MIC3表达产物分子量为46 ku,且经Western-blot显示可被猪抗弓形虫免疫血清识别.
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| 关 键 词： | 刚地弓形虫 微线体 克隆 表达 |
Cloning and prokaryotic expression of MIC3 protein-encoding gene of Toxoplasma gondii GJS strain |
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| GOU Hui-tian,WANG Yan-hua,LI Wen-hui,LI Hang,FU Bao-quan,ZHANG De-lin.Cloning and prokaryotic expression of MIC3 protein-encoding gene of Toxoplasma gondii GJS strain[J].Journal of Gansu Agricultural University,2008,43(5). |
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| Authors: | GOU Hui-tian WANG Yan-hua LI Wen-hui LI Hang FU Bao-quan ZHANG De-lin |
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| Abstract: | A pair of primers was designed according to the sequence of MIC3 from GenBank.The gene fragment encoding MIC3 was amplified by PCR from the total DNA of Toxoplasma gondii GJS strain and cloned into the plasmid pMD-18T to create plasmid pMD-MIC3.The constructed recombinant plasmid was transferred into E.coli JM109 susceptible cell and identified by enzyme digestion,PCR and sequencing.Another pair of expression primers was designed,amplified fragment was subcloned into pET-30a to create plasmid pET-MIC3.The recombinant plasmids were transformed into E.coli BL21(DE3).The expression of the recombinant pET-MIC3 was induced by IPTG.Then expressed products were analyzed by SDS-PAGE and Western-blot.The results showed that the size of recombinant MIC3 protein was about 46 ku.It could be specifically recognized by serum against Toxoplasma gondii from pig. |
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| Keywords: | Toxoplasma gondii MIC3 clone expression |
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