钩端螺旋体Lig A基因的原核表达及抗原性分析
发布时间：2019-04-08 18:55
【摘要】： 钩端螺旋体病作为全球范围内一个重要的公共卫生问题,防治工作越来越引起公众的关注。Lig A是钩端螺旋体外膜蛋白的一种,国外学者已对Lig A的生物学性质及其作为诊断试剂和疫苗的可行性有了初步的研究。由于国内暂无关于LigA蛋白的相关报道,国外虽有学者进行相关的研究工作,但国内外流行的钩端螺旋体的种属和血清型不同。寻找灵敏、特异性良好的抗原,对于国内钩体感染的早期诊断有很大帮助。 用PCR法扩增哥本哈根型钩体LigA基因,与表达载体pET-30a连接,获得LigA重组表达质粒,转化BL21(DE3)宿主菌,IPTG诱导表达。表达产物经亲和层析纯化后,采用western blot和ELISA分析其抗原性。 通过原核表达可以获得高产量的重组蛋白,western blot和ELISA结果显示重组抗原在检测国内常见的15群15型钩端螺旋体阳性标准血清方面具有良好的免疫活性。可以通过原核表达哥本哈根型钩体LigA基因,获得的重组蛋白可应用于钩体病血清抗体的检测,在钩体病诊断及疫苗研究中具有潜在的应用价值。
[Abstract]:Leptospirosis, as an important public health problem in the world, has attracted more and more public attention. Lig A is a kind of Leptospira outer membrane protein. The biological properties of Lig A and its feasibility as diagnostic reagent and vaccine have been studied. Since there are no relevant reports about LigA protein in China, although some foreign scholars have carried out relevant research work, the species and serotypes of Leptospira are different at home and abroad. Finding a sensitive and specific antigen is helpful for the early diagnosis of Leptospira infection in China. The LigA gene of Leptospira Copenhagen was amplified by PCR and ligated with the expression vector pET-30a to obtain the recombinant expression plasmid of LigA. The recombinant expression plasmid was transformed into BL21 (DE3) host strain and induced by IPTG. The expressed product was purified by affinity chromatography, and its antigenicity was analyzed by western blot and ELISA. High yield recombinant protein, western blot and ELISA were obtained by prokaryotic expression. The results showed that the recombinant antigen had good immune activity in detecting the positive standard serum of Leptospira 15, which was common in China. Through prokaryotic expression of Copenhagen Leptospira LigA gene, the recombinant protein can be used in the detection of serum antibody against leptospirosis, and it has potential application value in the diagnosis and vaccine research of Leptospira leptospirosis.
【学位授予单位】：福建医科大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R377.5
本文编号：2454824
[Abstract]:Leptospirosis, as an important public health problem in the world, has attracted more and more public attention. Lig A is a kind of Leptospira outer membrane protein. The biological properties of Lig A and its feasibility as diagnostic reagent and vaccine have been studied. Since there are no relevant reports about LigA protein in China, although some foreign scholars have carried out relevant research work, the species and serotypes of Leptospira are different at home and abroad. Finding a sensitive and specific antigen is helpful for the early diagnosis of Leptospira infection in China. The LigA gene of Leptospira Copenhagen was amplified by PCR and ligated with the expression vector pET-30a to obtain the recombinant expression plasmid of LigA. The recombinant expression plasmid was transformed into BL21 (DE3) host strain and induced by IPTG. The expressed product was purified by affinity chromatography, and its antigenicity was analyzed by western blot and ELISA. High yield recombinant protein, western blot and ELISA were obtained by prokaryotic expression. The results showed that the recombinant antigen had good immune activity in detecting the positive standard serum of Leptospira 15, which was common in China. Through prokaryotic expression of Copenhagen Leptospira LigA gene, the recombinant protein can be used in the detection of serum antibody against leptospirosis, and it has potential application value in the diagnosis and vaccine research of Leptospira leptospirosis.
【学位授予单位】：福建医科大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R377.5
【共引文献】
相关期刊论文 前2条
1 李国波;吴小楠;;钩端螺旋体病实验室诊断方法研究进展[J];海峡预防医学杂志;2010年03期
2 李国波;杨英超;辛晓芳;王国治;吴小南;;钩端螺旋体LigA基因原核表达及抗原性分析[J];药物分析杂志;2010年06期
，本文编号：2454824
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