耐甲氧西林金黄色葡萄球菌femA调控基因的初步研究

发布时间：2018-04-11 04:26

  本文选题：耐甲氧西林金黄色葡萄球菌 + β-内酰胺酶　； 参考：《四川大学》2006年硕士论文

【摘要】：目的：探讨耐甲氧西林金黄色葡萄球菌的辅助基因femA是否存在调控基因，以进一步揭示其耐药机制。 方法：用MIC法筛选对苯唑西林的高耐株、低耐株、敏感株；用Nitrocephin纸片法筛选出不产β-内酰胺酶菌株；用PCR筛选出含mecA的菌株。将含mecA不产β-内酰胺酶的高耐株、低耐株、敏感株作为进一步研究材料。 PCR扩增femA及fmA5′上游基因；提取细菌总蛋白；生物素标记femA上游DNA序列作为探针；滞留电泳检测femA调控基因与蛋白质的相互作用。 结果：在198株金黄色葡萄球菌中对苯唑西林高度耐药不产β-内酰胺酶含mecA基因菌株有55株；对苯唑西林低度耐药不产β-内酰胺酶含mecA基因菌株有5株；滞留电泳检测对苯唑西林高度耐药不产β-内酰胺酶含mecA基因菌株均出现滞留条带，，即发现femA上游DNA序列与苯唑西林高度耐药不产β-内酰胺酶的菌株耐受相关蛋白有相互作用。 结论：耐甲氧西林金黄色葡萄球菌的辅助基因femd有调控基因存在的可能性。作用的机理可能是一种正反馈模式。
[Abstract]:Aim: to investigate the existence of regulatory gene in methicillin-resistant Staphylococcus aureus cogene femA in order to reveal its mechanism of drug resistance.Methods: the high and low resistant strains of oxacillin were screened by MIC, the strains without 尾 -lactamase were screened by Nitrocephin disk method, and the strains containing mecA were screened by PCR.High resistant strains, low resistant strains and sensitive strains containing mecA did not produce 尾 -lactamases.The upstream genes of femA and fmA5'were amplified by PCR; the total bacterial protein was extracted; the upstream DNA sequence of femA was labeled with biotin as a probe; and the interaction between femA regulatory gene and protein was detected by stranded electrophoresis.Results: among the 198 strains of Staphylococcus aureus, 55 strains with high resistance to oxacillin did not produce 尾 -lactamases and 5 strains with mecA gene did not produce 尾 -lactamases.Stranded electrophoresis was used to detect the presence of stranded bands of 尾 -lactamases with high resistance to oxacillin and no 尾 -lactamases. It was found that the upstream DNA sequence of femA interacted with the tolerance related proteins of those strains that were not highly resistant to oxacillin producing 尾 -lactamases.Conclusion: the cogene femd of methicillin-resistant Staphylococcus aureus has the possibility of regulating genes.The mechanism of action may be a positive feedback mode.
【学位授予单位】：四川大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R378

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