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| 弓形虫P30基因的克隆及其在E．coli中的融合表达 | |
| 引用本文： | 王素华,曲道峰,蔡渭明,杜爱芳.弓形虫P30基因的克隆及其在E．coli中的融合表达[J].检验检疫科学,2008,18(4):8-10. |
| 作者姓名： | 王素华 曲道峰 蔡渭明 杜爱芳 |
| 作者单位： | 1. 温州出入境检验检疫局,浙江,温州,325027 2. 浙江大学动物预防医学研究所,浙江,杭州,310029 3. 绍兴出入境检验检疫局,浙江,绍兴,312000 |
| 基金项目： | 国家质检总局资助项目(No.2006IK016)猪弓形虫快速检测和防治技术研究 |
| 摘 要： | 【目的】研究弓形虫P30基因的克隆及其在E．coli中的融合表达。【方法】参照弓形虫P30序列设计引物，应用PCR技术扩增出弓形虫RH株P30的部分基因，将扩增产物克隆到pUCm—T载体，测序正确后再亚克隆到质粒pET-30a中，得到重组质粒pET-30a—P30并将其转化大肠杆菌BL21，IPTG诱导表达，表达产物经SDS-PAGE和Westen-blot分析。【结果】P30基因在E．coli中得到了高效表达，表达的融合蛋白的分子量在30kDa附近，可以被鼠抗Toxoplasma gondii阳性血清特异性识别。【结论】该研究为Toxoplasma gondii的检测和疫苗研制及综合防制奠定了基础。 |
| 关 键 词： | 弓形虫 P30基因 基因克隆 原核表达 |
CLONING AND PROKARYOTIC EXPRESSION OF P30 GENE OF Toxoplasma gondii IN E.coli |
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| WANG Suhua,QU Daofeng,CAIWeiming,DUAifang.CLONING AND PROKARYOTIC EXPRESSION OF P30 GENE OF Toxoplasma gondii IN E.coli[J].Inspection and Quarantine Science,2008,18(4):8-10. | |
| Authors: | WANG Suhua QU Daofeng CAIWeiming DUAifang |
| Institution: | WANG Suhua1,QU Daofeng2,CAI Weiming3,DU Aifang2( 1.Wenzhou Entry-3xit Inspection , Quarantine Bureau,Wenzhou,Zhejiang 325000,China,2.Institute of Preventive Veterinary Medicine,Zhejiang University,Hangzhou,Zhejiang 310029,3.Shaoxing Entry-Exit Inspection , Quarantine Bureau,Shaoxing,Zhejiang,312000 China,) |
| Abstract: | To study the cloning of P30 gene of Toxoplasma gondii,and its expression in E.coli.Specific primers were designed according to the sequence of P30 gene.P30 gene was obtained by PCR from RH Toxoplasma gondii,the DNA fragment of P30 gene was cloned into vector pUCm-T.After sequencing,the P30 gene was cloned into prokaryotic expression vector pET-30a plasmid.By induction of IPTG,and pET-30a-P30 plasmid was expressed.SDS-PAGE and Western blot assays indicated that the fusion proteins with about 30kDa were effic... |
| Keywords: | Toxoplasma gondii P30 gene Gene clone Prokaryotic expression |
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