本文關(guān)鍵詞： 銅綠假單胞菌 生物膜 MexAB-OprM MexR 亞胺培南 環(huán)丙沙星　出處：《青島大學(xué)》2009年碩士論文　論文類型：學(xué)位論文

【摘要】： 目的研究銅綠假單胞菌在浮游態(tài)、生物膜態(tài)及經(jīng)亞胺培南、環(huán)丙沙星誘導(dǎo)下,外排泵MexAB-OprM及其負(fù)性阻遏蛋白MexR表達(dá)情況。 方法用銅綠假單胞菌PA01在eppendorf管內(nèi)建立生物膜,通過結(jié)晶紫快速染色法證實(shí)生物膜的形成。用1/2MIC的亞胺培南、環(huán)丙沙星對銅綠假單胞菌浮游菌誘導(dǎo)4小時(shí);4MIC的亞胺培南、2MIC的環(huán)丙沙星對銅綠假單胞菌生物膜菌誘導(dǎo)12小時(shí),分別于生物膜形成的第3天及第5天,用實(shí)時(shí)熒光定量PCR的方法,測定銅綠假單胞菌外排泵MexAB-OprM及MexR的基因表達(dá)。 結(jié)果銅綠假單胞菌由浮游菌變?yōu)樯锬ぞ?第3天和第5天),其外排泵MexAB-OprM的表達(dá)水平明顯降低,其相對表達(dá)量最高相差近10倍,其差別存在統(tǒng)計(jì)學(xué)意義(P0.05),而生物膜菌第3天和第5天相比,盡管MexAB-OprM相對表達(dá)量相差近3倍,但差別無統(tǒng)計(jì)學(xué)意義(P0.05)。銅綠假單胞菌由浮游菌變?yōu)樯锬ぞ?第3天和第5天),阻遏蛋白MexR表達(dá)差別無統(tǒng)計(jì)學(xué)意義(P0.05)。生物膜菌第3天和第5天,其MexAB-OprM和MexR表達(dá)差別無統(tǒng)計(jì)學(xué)意義(P0.05)。在亞胺培南及環(huán)丙沙星誘導(dǎo)后,浮游菌和生物膜菌其基因MexAB-OprM、MexR表達(dá)差別均無統(tǒng)計(jì)學(xué)意義(P0.05)。 結(jié)論銅綠假單胞菌由浮游菌轉(zhuǎn)變?yōu)樯锬ぞ?外排泵MexAB-OprM表達(dá)明顯減少,MexR基因表達(dá)無影響。生物膜菌的第3天與第5天,其MexAB-OprM、MexR基因表達(dá)無差別。亞胺培南、環(huán)丙沙星對浮游菌及生物膜菌的MexAB-OprM、MexR均無誘導(dǎo)作用。
[Abstract]:Objective to study the expression of efflux pump MexAB-OprM and its negative repressor protein (MexR) in Pseudomonas aeruginosa in planktonic, biofilm, imipenem and ciprofloxacin. Methods the biofilm was established in the eppendorf tube with Pseudomonas aeruginosa PA01. The biofilm formation was confirmed by rapid staining of crystal violet, and 1 / 2 MIC imipenem was used. Ciprofloxacin induced phytoplankton of Pseudomonas aeruginosa for 4 hours. 4MIC imipenem 2MIC ciprofloxacin was induced to Pseudomonas aeruginosa biofilm for 12 hours, and real-time fluorescence quantitative PCR was used on the 3rd and 5th day of biofilm formation, respectively. The gene expression of MexAB-OprM and MexR in Pseudomonas aeruginosa efflux pump was determined. Results when Pseudomonas aeruginosa changed from phytoplankton to biofilm (on the 3rd and 5th day), the expression level of MexAB-OprM in efflux pump was significantly decreased, and the highest relative expression level was about 10 times. The difference was statistically significant (P 0.05), while the relative expression of MexAB-OprM was nearly three times higher than that of the third day and the fifth day of biofilm bacteria. But the difference was not statistically significant (P 0.05). Pseudomonas aeruginosa changed from phytoplankton to biofilm (day 3 and day 5). There was no significant difference in the expression of repressor protein MexR (P 0.05). The biofilm bacteria on the 3rd and 5th day. There was no significant difference in the expression of MexAB-OprM and MexR between them. After induction of imipenem and ciprofloxacin, the gene MexAB-OprM of phytoplankton and biofilm bacteria was found. The difference of MexR expression was not statistically significant (P 0.05). Conclusion after the transformation of Pseudomonas aeruginosa from phytoplankton to biofilm, the expression of MexAB-OprM in efflux pump has no effect on the expression of MexR gene, but on the 3rd and 5th day of biofilm bacteria. There was no difference in the expression of MexAB-Opr-MexR gene. Imipenem and ciprofloxacin did not induce MexAB-OprMexR gene expression in phytoplankton bacteria and biofilm bacteria.
【學(xué)位授予單位】：青島大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2009
【分類號】：R378.991

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