生物膜铜绿假单胞菌MexAB-OprM及MexR的表达

发布时间：2018-01-30 22:08

本文关键词： 铜绿假单胞菌生物膜 MexAB-OprM MexR 亚胺培南环丙沙星　出处：《青岛大学》2009年硕士论文　论文类型：学位论文

【摘要】： 目的研究铜绿假单胞菌在浮游态、生物膜态及经亚胺培南、环丙沙星诱导下,外排泵MexAB-OprM及其负性阻遏蛋白MexR表达情况。方法用铜绿假单胞菌PA01在eppendorf管内建立生物膜,通过结晶紫快速染色法证实生物膜的形成。用1/2MIC的亚胺培南、环丙沙星对铜绿假单胞菌浮游菌诱导4小时;4MIC的亚胺培南、2MIC的环丙沙星对铜绿假单胞菌生物膜菌诱导12小时,分别于生物膜形成的第3天及第5天,用实时荧光定量PCR的方法,测定铜绿假单胞菌外排泵MexAB-OprM及MexR的基因表达。结果铜绿假单胞菌由浮游菌变为生物膜菌后(第3天和第5天),其外排泵MexAB-OprM的表达水平明显降低,其相对表达量最高相差近10倍,其差别存在统计学意义(P0.05),而生物膜菌第3天和第5天相比,尽管MexAB-OprM相对表达量相差近3倍,但差别无统计学意义(P0.05)。铜绿假单胞菌由浮游菌变为生物膜菌后(第3天和第5天),阻遏蛋白MexR表达差别无统计学意义(P0.05)。生物膜菌第3天和第5天,其MexAB-OprM和MexR表达差别无统计学意义(P0.05)。在亚胺培南及环丙沙星诱导后,浮游菌和生物膜菌其基因MexAB-OprM、MexR表达差别均无统计学意义(P0.05)。结论铜绿假单胞菌由浮游菌转变为生物膜菌后,外排泵MexAB-OprM表达明显减少,MexR基因表达无影响。生物膜菌的第3天与第5天,其MexAB-OprM、MexR基因表达无差别。亚胺培南、环丙沙星对浮游菌及生物膜菌的MexAB-OprM、MexR均无诱导作用。
[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.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R378.991

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