本文選題：銅綠假單胞菌　切入點(diǎn)：吩嗪　出處：《西北大學(xué)》2008年碩士論文　論文類型：學(xué)位論文

【摘要】： 銅綠假單胞菌(Pseudomonas aeruginosa)是一個(gè)分布廣泛的機(jī)會(huì)致病菌[1],在燒傷病人、器官移植病人、免疫力低下病人,如接受化學(xué)治療的癌癥病人和艾滋病病人中,它幾乎可以感染人體的任何組織并常常危及患者生命。它對(duì)許多抗生素具備很高的內(nèi)在抗藥性,并可產(chǎn)生許多致病因子,如堿性蛋白酶(alkalineprotease)和吩嗪(phenazine)等,是難以治療的病原菌。 綠膿菌素作為吩嗪的一種衍生物,不僅是一種致病因子,更重要的是作為銅綠假單胞菌中的一種信號(hào)分子。因此,研究吩嗪合成的調(diào)節(jié)途徑對(duì)于了解銅綠假單胞菌的致病機(jī)制和微生物群體中不同細(xì)菌間的相互作用具有重要意義。 本文運(yùn)用轉(zhuǎn)座子突變方法,對(duì)含有報(bào)道質(zhì)粒miniCTX-phzAl的銅綠假單胞菌進(jìn)行轉(zhuǎn)座子隨機(jī)突變,篩選影響吩嗪合成的調(diào)節(jié)突變體。篩選到了6個(gè)影響吩嗪合成的突變體,并對(duì)這6個(gè)突變體進(jìn)行了鑒定,確定突變體中轉(zhuǎn)座子插入位點(diǎn),從而確定轉(zhuǎn)座子所破壞的基因。并根據(jù)生物信息分析結(jié)果,推測(cè)其功能。研究結(jié)果為了解銅綠假單胞菌致病因子的調(diào)節(jié)機(jī)理提供了新的數(shù)據(jù)和線索。 同時(shí),對(duì)一個(gè)菌落和顏色發(fā)生明顯變化的突變體進(jìn)行的深入研究表明,該轉(zhuǎn)座突變的基因是銅綠假單胞菌的一個(gè)新轉(zhuǎn)錄調(diào)節(jié)基因。并通過(guò)基因敲除的方法驗(yàn)證了該基因的功能。 采用基因敲除的方法,構(gòu)建了負(fù)責(zé)ABC轉(zhuǎn)運(yùn)系統(tǒng)的兩個(gè)基因PA0602和PA4594基因的突變體。研究這兩個(gè)基因是否參與低抑制濃度四環(huán)素對(duì)吩嗪合成基因phzAl的調(diào)節(jié)。探索了這兩個(gè)基因與群體感應(yīng)體調(diào)節(jié)系統(tǒng)和PQS系統(tǒng)之間一定的關(guān)系。
[Abstract]:Pseudomonas aeruginosa (Pseudomonas aeruginosa) is a widely distributed opportunistic pathogen in burn patients, organ transplant patients, immunocompromised patients, such as cancer patients receiving chemotherapy and AIDS patients. It can infect almost any human tissue and often endanger the life of patients. It has high intrinsic resistance to many antibiotics and can produce many pathogenic factors, such as alkaline protease, phenazine and phenazine, which are difficult to treat. As a derivative of phenazine, Pseudomonas aeruginosa is not only a pathogenic factor, but also a signaling molecule in Pseudomonas aeruginosa. It is important to study the regulatory pathway of phenazine synthesis for understanding the pathogenic mechanism of Pseudomonas aeruginosa and the interaction between different bacteria in microbial population. In this paper, transposon random mutation of Pseudomonas aeruginosa containing reporter plasmid miniCTX-phzAl was carried out by transposon mutation method to screen regulatory mutants affecting phenazine synthesis. Six mutants affecting phenazine synthesis were screened. The six mutants were identified and the transposon insertion sites were determined to determine the genes destroyed by transposons. The results provide new data and clues for understanding the regulatory mechanism of the pathogenic factors of Pseudomonas aeruginosa. At the same time, an in-depth study of a mutant with significant changes in colony and color showed that, The translocation mutant gene is a new transcriptional regulator of Pseudomonas aeruginosa and its function is verified by gene knockout. Using gene knockout, The mutants of two genes, PA0602 and PA4594, responsible for the ABC transport system, were constructed. Whether the two genes were involved in the regulation of phenazine synthesis gene phzAl by low inhibitory concentration tetracycline was studied. There is a certain relationship between the regulating system and the PQS system.
【學(xué)位授予單位】：西北大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2008
【分類號(hào)】：R378

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