蟾酥对金黄色葡萄球菌抑菌作用及机理的初步研究-生物学专业论文.docxVIP

中文摘要 中文摘要 万方数据 万方数据 中文摘要 本实验采用 2,3,5-氯化三苯基四氮唑(TTC)染色、菌液电导率测定、RT-PCR 实验、微孔板培养和结晶紫染色定量、激光共聚焦显微成像系统观察，分别从菌 体生长状态、生长曲线、细胞膜透、细菌内转肽酶(femB)基因的 RNA 转录水平 和细菌生物被膜形成等方面对蟾酥对金黄色葡萄球菌作用机理进行初步研究；研 究结果表明蟾酥对金黄色葡萄球菌最小抑菌浓度(MIC)为 2mg/mL，最小杀菌浓 度(MBC)为 2.5mg/mL；细胞膜通透性与对照组相比显著提高；RT-PCR 实验表明， 金黄色葡萄球菌的 femB 基因的 mRNA 转录水平随蟾酥溶液作用时间的增加而 逐渐降低，作用 9h 时 fenB 基因的 mRNA 水平为正常菌的 0.53 倍；蟾酥对金黄 色葡萄球菌生物被膜形成有效抑制浓度为 1.25mg/mL；蟾酥通过干扰金黄色葡萄 球菌细胞壁的合成以及抑制其生物被摸的形成而抑制金黄色葡萄球菌生长。 关键词 蟾酥 ；金黄色葡萄球菌 ；抑菌机理 ；生物被膜 I 牡丹江师范学院硕士学位论文 牡丹江师范学院硕士学位论文 Abstract In this work, preliminary studies have been carried out on the mechanism of toad venom against Staphylococcus aureus in aspect of cell growth status, growth curve, cell membrane permeability and RNA transcription level of bacterial GGT femB gene. Study procedure contains 2,3,5-triphenyl tetrazolium chloride (TTC) Staining, bacteria conductivity measurement, RT-PCR experiments, culture microplates, crystal violet staining method for quantifying and observation with laser confocal microscopy imaging system. Results show that the minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) of toad venom against Staphylococcus aureus are 2mg/mL and 2.5mg/mL, respectively. The membrane permeability significantly increased compared with the control group. RT-PCR experiments showed a decline in mRNA levels of Staphylococcus aureus femB gene with treatment time increasing, while the mRNA levels of femB gene was 0.53 times of the normal bacteria after 9 hours. Besides, along with the suppression on the bacterial growth, an inhibiting effect can be also observed for toad venom on the biofilm formation of Staphylococcus aureus with an effective inhibitory concentration of 1.25mg/mL. Toad venom can suppress growth of Staphylococcus aureus by obstructing cytoderm synthesis and biofilm formatio. Keywords: toad venom; Staphylococcus aureus; antibacterial mechanism; biofilm II 目 目 录 目 录 HYPERLINK \l _bookmark0 中文摘要.....................................