Gene expression and protein analysis of the antibiotic extrusion pump in Pseudomonas aeruqinos

铜绿假单胞菌抗生素挤出泵基因表达及蛋白分析

• 批准号: 07457073
• 负责人: NAKAE Taiji
• 金额: $ 3.58万
• 依托单位: Tokai University School of Medicine
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07457073/
• 关键词: antibiotics; druq resistance; efflux; Pseudomonas; extrusion pump; cell membrane; MDR; membrane protein; Pseudomonas; 薬剤排出

We cloned the gene (s) associated with multiantibiotic resistance in Pseudomonas aeruginosa from a mutant with elevated drug extrusion. The strain harboring cloned gene accumulated less amount of ofloxacin than the strain without the clone. By using Southern and northern blot analyzes, we investigated whether multiantibiotic resistance is caused by gene amplification or overexpression. We found that all our multiantibiotic resistant mutants isolated earlier overexpressed mRNA homologous to the cloned gene (s). Overexpresssions of the 41 KDa and 110 KDa inner-and the 50 KDa outer membrane proteins were detected. Other multiantibiotic resistant mutants including the nalB mutant overexpressed the mRNA and the membrane proteins, but the nfxB and nfxC mutants did a little. We concluded that low antibiotic accumulation in the multiantibiotic resistant mutants is attributable to overexpression of the antibiotic extrusion machinery.

我们克隆了与铜绿假单胞菌多抗生素耐药性相关的基因，该基因来自一个药物挤压升高的突变体。含有克隆基因的菌株比没有克隆的菌株积累的氧氟沙星量少。通过Southern和northern blot分析，我们研究了多种抗生素耐药是由基因扩增还是过度表达引起的。我们发现所有早期分离的多抗生素耐药突变体都过表达与克隆基因同源的mRNA。检测到41 KDa和110 KDa的膜内蛋白和50 KDa的膜外蛋白过表达。包括nalB突变体在内的其他多抗生素耐药突变体过度表达mRNA和膜蛋白，但nfxB和nfxC突变体则略微过度表达。我们得出结论，在多抗生素耐药突变体中，抗生素积累较低是由于抗生素挤出机制的过度表达。

项目成果

Yoneyama.H.and Nakae.T: "Protein C of the outer membarane of Pseudomonas aeruginosa is the copper regulated channel protein." Microbiology. 142. 2137-2144 (1996)

Yoneyama.H. 和 Nakae.T：“铜绿假单胞菌外膜的蛋白 C 是铜调节通道蛋白。”

Yoshihara,E.,Gotoh,N.,Nishino,T.,Nakae,T.: "Protein D2 porin of Psudomonas aeruginosa outer membrane bears the protease activity." FEBS Letters. 394. 179-182 (1996)

Yoshihara,E.、Gotoh,N.、Nishino,T.、Nakae,T.：“铜绿假单胞菌外膜的蛋白质 D2 孔蛋白具有蛋白酶活性。”

Nakae.T.: "Characterization of novel hemagglutinin of diarrhea-associated Escherichia coli tha has characateristics of diffusely adhering E.coli and Enteroaggregative E.coli." Infect.Immun.64. 3694-3702 (1996)

Nakae.T.：“腹泻相关大肠杆菌的新型血凝素特征，其具有弥漫性粘附大肠杆菌和肠聚集性大肠杆菌的特征。”

Nakae,T.: "Role of membrane permeability in determining antibiotic resistance inPseudomonas aeruginosa." Microbiol.Immunol.39. 211-229 (1995)

Nakae,T.：“膜通透性在确定铜绿假单胞菌抗生素耐药性中的作用。”

Ikigai.H..Akatuka.A..Tsuiivama.H..Nakae.T.: "Mechanism of membrane damage by EITor hemolysin of Vibrio cholerae O1." Infect.Immunity. 64. 2968-2973 (1996)

Ikigai.H..Akatuka.A..Tsuiivama.H..Nakae.T.：“霍乱弧菌 O1 的 EITor 溶血素造成膜损伤的机制。”