本文關(guān)鍵詞： 鉤端螺旋體 基因組學(xué) 表面暴露蛋白質(zhì)組 毒力因子 硫醇過(guò)氧化物酶　出處：《上海交通大學(xué)》2015年博士論文　論文類型：學(xué)位論文

【摘要】：鉤端螺旋體病是由致病性鉤端螺旋體引起的一種全球范圍內(nèi)的人獸共患病,感染鉤端螺旋體可引起黃疸、發(fā)熱、出血以及急性的多器官衰竭等危害,其嚴(yán)重威脅著人類的健康,與此同時(shí)也對(duì)畜牧業(yè)造成了極大的損失。由于鉤體特殊的進(jìn)化地位以及有效遺傳操作系統(tǒng)的缺乏,人們對(duì)于鉤體及鉤體病的研究進(jìn)展一直十分緩慢,鉤體的主要致病因子及致病機(jī)制仍不明確,亦缺乏相應(yīng)的診斷及疫苗靶標(biāo)。直到2003年第一株鉤體全基因組測(cè)序數(shù)據(jù)的報(bào)道,基于全基因組測(cè)序技術(shù)的相應(yīng)研究不斷為人們對(duì)于鉤體生理、代謝以及致病機(jī)制的研究和認(rèn)識(shí)提供新的方法和視野。本論文基于對(duì)我國(guó)致病性鉤體主要血清群代表株的基因組測(cè)序研究,初步探討了鉤體不同血清群泛基因組的組成,并以核心基因組為基礎(chǔ)進(jìn)一步分析了這些血清群間的進(jìn)化關(guān)系。另外,通過(guò)454焦磷酸測(cè)序并結(jié)合Sanger測(cè)序技術(shù),完成了我國(guó)鉤體疫苗株56610(401)株的全基因組測(cè)序研究,鑒定出了其染色體外環(huán)狀質(zhì)粒的存在以及特異的O抗原編碼區(qū)域。結(jié)合課題組已經(jīng)完成全基因組測(cè)序的另一株鉤體疫苗菌株56603(Gui44)株以及國(guó)際上已公布的6株鉤體(4株強(qiáng)毒及2株腐生型鉤體)全基因組測(cè)序數(shù)據(jù),我們也對(duì)8株不同毒力的鉤體全基因組進(jìn)行了比較分析。通過(guò)對(duì)致病性菌株特有基因的篩選,預(yù)測(cè)到20個(gè)潛在的核心毒力編碼基因。在全基因組測(cè)序的基礎(chǔ)上,我們完成了問(wèn)號(hào)鉤體56601株、56603(Gui44)株及56610(401)株的表面暴露蛋白質(zhì)組質(zhì)譜鑒定,結(jié)合優(yōu)化的表面暴露蛋白預(yù)測(cè)策略共篩選的81個(gè)核心表面暴露蛋白、61個(gè)非必需表面暴露蛋白以及122個(gè)特異表面暴露蛋白。另外,我們?cè)诜置诘鞍踪|(zhì)組內(nèi)篩選到一個(gè)保守存在的蛋白-硫醇過(guò)氧化物酶。通過(guò)實(shí)驗(yàn)證實(shí),由基因LA0862編碼的鉤體硫醇過(guò)氧化物酶可以通過(guò)非典型的分泌途徑分泌到胞外,在鉤體與宿主相互作用的過(guò)程中,保護(hù)鉤體免受外源過(guò)氧化物壓力的威脅。重組表達(dá)的鉤體硫醇過(guò)氧化物酶刺激產(chǎn)生的抗體與致病性鉤體具有較好的交叉免疫反應(yīng),鉤體硫醇過(guò)氧化物酶和鉤體免疫的動(dòng)物血清也可以發(fā)生反應(yīng),這說(shuō)明LA0862可以做為鉤體的疫苗及診斷的靶點(diǎn)。
[Abstract]:Leptospirosis is a worldwide zoonosis caused by pathogenic leptospirosis. Infection with leptospirosis can cause jaundice, fever, bleeding and acute multiple organ failure. At the same time, it has caused great losses to animal husbandry. Due to the special evolutionary status of leptospirosis and the lack of effective genetic operating system, the progress of research on leptospirosis and leptospirosis has been very slow. The main pathogenic factors and pathogenetic mechanism of Leptospira were still unclear, and the corresponding diagnosis and vaccine target were lacking. Until 2003, the first Leptospira genome sequencing data were reported. The corresponding research based on the whole genome sequencing technology has been used for the study of Leptospira physiology. The study and understanding of metabolism and pathogenesis provide a new method and perspective. Based on the genome sequencing of the main serogroup representative strains of leptospira in China, the composition of pangenome of different serogroups of Leptospira was preliminarily discussed. Based on the core genomes, the evolutionary relationships among these serogroups were further analyzed. In addition, the whole genome sequencing of Chinese leptospirosis vaccine strain 56610 / 401 was completed by 454 pyrosequencing and Sanger sequencing. The exocyclic plasmids and the specific O antigen coding region were identified. Another Leptospira vaccine strain 56603 Gui44, which has been sequenced by our research group, and 6 Leptospira Leptospira strains published internationally, have been identified. Complete genome sequencing data of two strains of saprophytic leptospirosis. We also compared and analyzed the whole genome of eight different virulence leptospira strains. By screening the specific genes of pathogenic strains, we predicted 20 potential core virulence coding genes. We have completed the surface exposure proteome mass spectrometry identification of 56601 strains of Leptospira interrogans, 56603, Gui44, and 5661010401. A total of 81 core surface exposure proteins, 61 non-essential surface exposure proteins and 122 specific surface exposure proteins were screened in combination with the optimized surface exposure protein prediction strategy. We screened a conserved protein-mercaptoperoxidase (mercapto peroxidase) from the secretory proteome. It was proved by experiments that the leptospirase encoded by the gene LA0862 can be secreted to the extracellular via atypical secretory pathway. During the interaction between Leptospira and host, Leptospira was protected from the threat of exogenous peroxidase. The antibody produced by recombinant Leptospira mercaptoperoxidase stimulated by Leptospira had a good cross-immune response to pathogenic Leptospira. Leptospira mercaptoperoxidase and Leptospira immunized animal serum can also react, which suggests that LA0862 can be used as a vaccine and a diagnostic target for leptospirosis.
【學(xué)位授予單位】：上海交通大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2015
【分類號(hào)】：R514.4

【相似文獻(xiàn)】

相關(guān)期刊論文 前8條

1 張芳,李文俊,楊宏亮,何平,郭曉奎,姜敘誠(chéng);問(wèn)號(hào)鉤體鞭毛蛋白相關(guān)基因分析[J];中國(guó)人獸共患病雜志;2004年12期

2 耿燕,郭曉奎,張怡軒,繆有剛,何建勇,趙國(guó)屏;鉤體LPS生物合成與裝配途徑及相關(guān)基因分析[J];中國(guó)人獸共患病雜志;2004年01期

3 楊宏亮,李文俊,姜敘誠(chéng),郭曉奎;問(wèn)號(hào)鉤體粘附侵襲相關(guān)基因特征分析[J];微生物學(xué)雜志;2004年03期

4 鮑朗,熊秋芳,張明,伍衛(wèi)華,萬(wàn)波,李勝富;應(yīng)用低嚴(yán)格度-單引物聚合酶鏈反應(yīng)對(duì)問(wèn)號(hào)鉤體中國(guó)參考株的基因多態(tài)性分析[J];華西醫(yī)科大學(xué)學(xué)報(bào);1997年02期

5 顧艷,楊宏亮,李文俊,姜敘誠(chéng),郭曉奎;問(wèn)號(hào)鉤體Ⅱ型分泌系統(tǒng)相關(guān)基因特征分析[J];中國(guó)生物制品學(xué)雜志;2004年05期

6 趙莉,蔣秀高,聶一新,肖玉春,徐建國(guó);鉤端螺旋體毒力相關(guān)基因的分布特點(diǎn)分析[J];中華流行病學(xué)雜志;2003年12期

7 李亞非,戴保民,趙慧業(yè);針對(duì)問(wèn)號(hào)鉤體賴型外膜的抗原分析[J];華西醫(yī)科大學(xué)學(xué)報(bào);1993年02期

8 ;[J];;年期

相關(guān)博士學(xué)位論文 前10條

1 朱威南;問(wèn)號(hào)鉤體56609株基因組特征及鉤體轉(zhuǎn)化系統(tǒng)建立[D];上海交通大學(xué);2014年

2 葛玉梅;問(wèn)號(hào)鉤端螺旋體鋅離子依賴基質(zhì)金屬蛋白酶致病作用及機(jī)制[D];浙江大學(xué);2016年

3 莊緒冉;基于組學(xué)研究對(duì)致病性鉤體毒力因子及免疫靶標(biāo)的分析[D];上海交通大學(xué);2015年

4 張成林;感染狀態(tài)下問(wèn)號(hào)鉤體脂多糖合成、修飾及其調(diào)控機(jī)制的研究[D];浙江大學(xué);2012年

5 金丹丹;問(wèn)號(hào)鉤端螺旋體誘導(dǎo)小鼠巨噬細(xì)胞凋亡及其信號(hào)傳導(dǎo)機(jī)制的研究[D];浙江大學(xué);2009年

6 李世軍;問(wèn)號(hào)鉤端螺旋體誘導(dǎo)細(xì)胞凋亡及吞噬細(xì)胞內(nèi)存活與繁殖分子機(jī)制研究[D];浙江大學(xué);2009年

7 KASSEGNE Kokouvi;鉤端螺旋體膠原蛋白酶致病作用及機(jī)制研究[D];浙江大學(xué);2012年

8 黃莉莉;問(wèn)號(hào)鉤端螺旋體犬群犬型Gui44株基因組與蛋白質(zhì)組的研究[D];上海交通大學(xué);2012年

9 董海艷;線粒體相關(guān)途徑在問(wèn)號(hào)鉤端螺旋體誘導(dǎo)巨噬細(xì)胞凋亡中的作用以及致病性鉤端螺旋體OmpL1特點(diǎn)的研究[D];浙江大學(xué);2009年

10 胡瑋琳;鉤端螺旋體誘導(dǎo)巨噬細(xì)胞凋亡機(jī)制及其抗原表位免疫保護(hù)作用研究[D];浙江大學(xué);2014年

相關(guān)碩士學(xué)位論文 前3條

1 張磊;問(wèn)號(hào)鉤端螺旋體粘附侵襲相關(guān)基因mce功能鑒定[D];浙江大學(xué);2010年

2 辛?xí)躁?yáng);鉤端螺旋體毒性蛋白VapC核酸酶活性及其對(duì)宿主細(xì)胞毒性作用的研究[D];浙江大學(xué);2012年

3 邱曉楓;基于問(wèn)號(hào)鉤端螺旋體rLipL32/1-LipL21-OmpL1/2融合抗原ELISAs的建立及應(yīng)用[D];浙江大學(xué);2009年

，

本文編號(hào)：1499330