| 猪链球菌谷氨酰胺tRNA合成酶的原核表达产物的小鼠免疫试验 |
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| 引用本文： | 胡睿铭,赵战勤,李伦勇,李增强,汤细彪,段龙川,吴斌.猪链球菌谷氨酰胺tRNA合成酶的原核表达产物的小鼠免疫试验[J].微生物学报,2010,50(3):418-422. |
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| 作者姓名： | 胡睿铭 赵战勤 李伦勇 李增强 汤细彪 段龙川 吴斌 |
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| 作者单位： | 华中农业大学农业微生物学国家重点实验室,武汉,430070 |
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| 基金项目： | 国家公益性农业行业科研专项(200803020)；国家科技攻关项目的资助 (2006BAD06A18-2) |
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| 摘 要： | 【目的】猪链球菌谷氨酰tRNA合成酶(Glutamyl tRNA Synthetase,GtS)是一种催化谷氨酰胺与对应的tRNA发生酯化反应形成谷氨酰tRNA的蛋白酶。本研究通过小鼠免疫保护力实验来评价Gts的免疫原性。【方法与结果】使用clustalX对GenBank中链球菌属的GtS进行同源性分析,结果显示GtS的氨基酸序列在链球菌属内同源性达到95%以上。利用原核表达系统在大肠杆菌BL21中表达(His)6-GtS融合蛋白。SDS-PAGE和Western blot结果表明该融合蛋白在BL21菌株中高效表达,并且具有良好的免疫原性。融合蛋白经过组氨酸标签纯化试剂盒(Novagen)纯化后获得纯度为93.3%、浓度为433μg/mL的重组蛋白rGtS。在小鼠免疫保护试验中,使用rGtS混合弗氏佐剂免疫Balb/c小鼠。免疫过后用4倍LD50剂量的SC19菌株(1.2×109CFU)攻毒,rGtS免疫组小鼠的存活率达50%(4/8),高于空载体对照(1/8)。【结论】本研究证实rGtS融合蛋白具有良好免疫原性,能够提供部分的保护,是一种非常有潜力的亚单位疫苗候选蛋白。
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| 关 键 词： | 关键词：猪链球菌，谷氨酰胺tRNA合成酶，免疫原性 |
| 收稿时间： | 2009/10/8 0:00:00 |
| 修稿时间： | 2009/12/8 0:00:00 |
Prokaryotic expression of Glutamyl tRNA Synthetase of Streptococcus suis and its immunization experiment |
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| Ruiming Hu,Zhangqing Zhao,Lunyong Li,Zengqiang Li,Xibiao Tang,Longchuan Duan and Bin Wu.Prokaryotic expression of Glutamyl tRNA Synthetase of Streptococcus suis and its immunization experiment[J].Acta Microbiologica Sinica,2010,50(3):418-422. |
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| Authors: | Ruiming Hu Zhangqing Zhao Lunyong Li Zengqiang Li Xibiao Tang Longchuan Duan and Bin Wu |
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| Institution: | State Key Laboratory of Agriculture Microbiology, Huazhong Agricultural University, Wuhan 430070, China;State Key Laboratory of Agriculture Microbiology, Huazhong Agricultural University, Wuhan 430070, China;State Key Laboratory of Agriculture Microbiology, Huazhong Agricultural University, Wuhan 430070, China;State Key Laboratory of Agriculture Microbiology, Huazhong Agricultural University, Wuhan 430070, China;State Key Laboratory of Agriculture Microbiology, Huazhong Agricultural University, Wuhan 430070, China;State Key Laboratory of Agriculture Microbiology, Huazhong Agricultural University, Wuhan 430070, China;State Key Laboratory of Agriculture Microbiology, Huazhong Agricultural University, Wuhan 430070, China |
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| Abstract: | Objective]Glutamyl tRNA Synthetase (Gts) is a protease which catalyzes esterification between tRNA and glutamine. The immunogenicity of Gts was evaluated through immunization and challenge experiment. Methods and results] We cloned gts from the genome of SC19,and inserted it into prokaryotic expression plasmid pET28a-gts. The recombinant vector was transformed into E. coli BL21. Induced by IPTG,one 58 kD protein,was expressed and purified by using Ni~+-NTA column (Novagen). The purity of rGtS was 93.3% an... |
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| Keywords: | Keywords: Streptococcus suis Glutamyl tRNA Synthetase immunogenicitys |
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