本文選題：創(chuàng)傷弧菌　切入點(diǎn)：溶細(xì)胞素　出處：《浙江大學(xué)》2006年碩士論文　論文類型：學(xué)位論文

【摘要】：背景和目的 創(chuàng)傷弧菌(Vibrio vulnificus)是一種有莢膜的革蘭陰性嗜鹽弧菌,存在于海水及海產(chǎn)品中,由Farmer等于1979年首先報(bào)道。該菌具有強(qiáng)烈毒性,主要引起原發(fā)性敗血癥和嚴(yán)重的創(chuàng)口感染。敗血癥多與生食含菌的牡蠣等貝類海產(chǎn)品有關(guān),尤其好發(fā)于肝硬化、血色素沉著癥、地中海貧血等體內(nèi)荷鐵量過多的人群中,病死率超過50%。皮膚創(chuàng)口受海水中的創(chuàng)傷弧菌感染,常于24小時(shí)內(nèi)迅速導(dǎo)致嚴(yán)重的組織壞死。創(chuàng)傷弧菌的確切致病機(jī)理至今尚未完全明了。溶細(xì)胞素是由結(jié)構(gòu)基因vvhA編碼的一種相對(duì)分子質(zhì)量為50851的細(xì)胞外蛋白質(zhì),具有水溶性,由大多數(shù)致病菌株產(chǎn)生,能溶解哺乳動(dòng)物紅細(xì)胞及中國倉鼠卵巢(CHO)細(xì)胞,是創(chuàng)傷弧菌向胞外釋放的唯一細(xì)胞毒素,一直被認(rèn)為是創(chuàng)傷弧菌重要的致病因子,且因其具有穿孔特性而頗受關(guān)注。目前對(duì)創(chuàng)傷弧菌溶細(xì)胞素的細(xì)胞毒性機(jī)制已有廣泛研究和報(bào)道,但其確切機(jī)制尚未完全明了。 為了給今后研究探討創(chuàng)傷弧菌溶細(xì)胞素的致病機(jī)制以及制備創(chuàng)傷弧菌基因工程疫苗和免疫診斷試劑盒等奠定實(shí)驗(yàn)基礎(chǔ),本研究在我們實(shí)驗(yàn)室以往工作基礎(chǔ)上,用工程菌BL21DE3~(pET-32a(+)-vvhA)表達(dá)vvhA融合蛋白,對(duì)其進(jìn)行純化、復(fù)性;通過臺(tái)盼藍(lán)染料排斥試驗(yàn)檢測vvhA融合蛋白對(duì)細(xì)胞活力的影響;分別以2,
[Abstract]:Background and objective Vibrio vulnificus (Vibrio vulnificus) is a gram-negative halophilic vibrio with capsule, which is found in seawater and seafood, first reported by Farmer in 1979. Mainly causing primary sepsis and severe wound infection. Septicemia is associated with shellfish such as oyster, which is raw and contains bacteria, especially in people with liver cirrhosis, hemochromatosis, thalassemia and other people with excessive amounts of iron in their bodies. The mortality rate is more than 50%. The skin wound is infected by Vibrio vulnificus in seawater. The exact pathogenicity of Vibrio vulnificus is not fully understood. Cytolysin is an extracellular protein with a relative molecular weight of 50851 encoded by the structural gene vvhA, which is water-soluble. Produced by most pathogenic bacteria strains and capable of dissolving mammalian erythrocytes and Chinese hamster ovary Cho cells, it is the only cytotoxin released by Vibrio vulnificus to extracellular, and has been regarded as an important pathogenic factor of Vibrio vulnificus. The cytotoxicity mechanism of Vibrio vulnificus cytolysin has been widely studied and reported, but its exact mechanism has not been fully understood. In order to lay an experimental foundation for further study on the pathogenicity of vibrio vulnificus lytic cytokine and the preparation of genetic engineering vaccine and immunological diagnostic kit of Vibrio vulnificus, this study was based on the previous work in our laboratory. The vvhA fusion protein was expressed by BL21DE3, pET-32a (TvvhA), purified and renatured. The effect of vvhA fusion protein on cell viability was detected by trypan blue dye exclusion assay.
【學(xué)位授予單位】：浙江大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2006
【分類號(hào)】：R378

【共引文獻(xiàn)】

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3 趙旭鴻;陸森泉;;創(chuàng)傷弧菌溶細(xì)胞素細(xì)胞毒性機(jī)制的研究進(jìn)展[J];微生物與感染;2006年01期

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本文編號(hào)：1602932