【摘要】： 鉤端螺旋體病作為全球范圍內(nèi)一個(gè)重要的公共衛(wèi)生問(wèn)題,防治工作越來(lái)越引起公眾的關(guān)注。Lig A是鉤端螺旋體外膜蛋白的一種,國(guó)外學(xué)者已對(duì)Lig A的生物學(xué)性質(zhì)及其作為診斷試劑和疫苗的可行性有了初步的研究。由于國(guó)內(nèi)暫無(wú)關(guān)于LigA蛋白的相關(guān)報(bào)道,國(guó)外雖有學(xué)者進(jìn)行相關(guān)的研究工作,但國(guó)內(nèi)外流行的鉤端螺旋體的種屬和血清型不同。尋找靈敏、特異性良好的抗原,對(duì)于國(guó)內(nèi)鉤體感染的早期診斷有很大幫助。 用PCR法擴(kuò)增哥本哈根型鉤體LigA基因,與表達(dá)載體pET-30a連接,獲得LigA重組表達(dá)質(zhì)粒,轉(zhuǎn)化BL21(DE3)宿主菌,IPTG誘導(dǎo)表達(dá)。表達(dá)產(chǎn)物經(jīng)親和層析純化后,采用western blot和ELISA分析其抗原性。 通過(guò)原核表達(dá)可以獲得高產(chǎn)量的重組蛋白,western blot和ELISA結(jié)果顯示重組抗原在檢測(cè)國(guó)內(nèi)常見(jiàn)的15群15型鉤端螺旋體陽(yáng)性標(biāo)準(zhǔn)血清方面具有良好的免疫活性�？梢酝ㄟ^(guò)原核表達(dá)哥本哈根型鉤體LigA基因,獲得的重組蛋白可應(yīng)用于鉤體病血清抗體的檢測(cè),在鉤體病診斷及疫苗研究中具有潛在的應(yīng)用價(jià)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.
【學(xué)位授予單位】：福建醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2010
【分類號(hào)】：R377.5

【共引文獻(xiàn)】

相關(guān)期刊論文 前2條

1 李國(guó)波;吳小楠;;鉤端螺旋體病實(shí)驗(yàn)室診斷方法研究進(jìn)展[J];海峽預(yù)防醫(yī)學(xué)雜志;2010年03期

2 李國(guó)波;楊英超;辛?xí)苑?王國(guó)治;吳小南;;鉤端螺旋體LigA基因原核表達(dá)及抗原性分析[J];藥物分析雜志;2010年06期

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本文編號(hào)：2454824