本文選題：耐甲氧西林金黃色葡萄球菌 + β-內(nèi)酰胺酶��； 參考：《四川大學》2006年碩士論文

【摘要】：目的：探討耐甲氧西林金黃色葡萄球菌的輔助基因femA是否存在調(diào)控基因，以進一步揭示其耐藥機制。 方法：用MIC法篩選對苯唑西林的高耐株、低耐株、敏感株；用Nitrocephin紙片法篩選出不產(chǎn)β-內(nèi)酰胺酶菌株；用PCR篩選出含mecA的菌株。將含mecA不產(chǎn)β-內(nèi)酰胺酶的高耐株、低耐株、敏感株作為進一步研究材料。 PCR擴增femA及fmA5′上游基因；提取細菌總蛋白；生物素標記femA上游DNA序列作為探針；滯留電泳檢測femA調(diào)控基因與蛋白質(zhì)的相互作用。 結(jié)果：在198株金黃色葡萄球菌中對苯唑西林高度耐藥不產(chǎn)β-內(nèi)酰胺酶含mecA基因菌株有55株；對苯唑西林低度耐藥不產(chǎn)β-內(nèi)酰胺酶含mecA基因菌株有5株；滯留電泳檢測對苯唑西林高度耐藥不產(chǎn)β-內(nèi)酰胺酶含mecA基因菌株均出現(xiàn)滯留條帶，，即發(fā)現(xiàn)femA上游DNA序列與苯唑西林高度耐藥不產(chǎn)β-內(nèi)酰胺酶的菌株耐受相關(guān)蛋白有相互作用。 結(jié)論：耐甲氧西林金黃色葡萄球菌的輔助基因femd有調(diào)控基因存在的可能性。作用的機理可能是一種正反饋模式。
[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.
【學位授予單位】：四川大學
【學位級別】：碩士
【學位授予年份】：2006
【分類號】：R378

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