Abstract 1
中文摘要 2
前言 3
鮑氏不動桿菌 3
雙分子調控系統 4
鮑氏不動桿菌的雙分子調控系統 4
研究源起與目的 7
材料與方法 8
一、材料 8
1. 質體、菌種以及引子 8
2. 大蠟蛾 (Galleria mellonella)生長環境以及成長史 8
3. 藥品及試劑 8
4. 儀器與器材 8
二、方法 9
1. Luria-Bertani (LB)培養基製備 9
2. 純化染色體 (Chromosome extraction) 9
3. DNA聚合酶鏈鎖反應 (Polymerase chain reaction, PCR) 10
4. 純化質體 (Plasmid extraction) 10
5. 限制酶作用 (Enzyme digestion) 11
6. DNA黏合作用 (Ligation) 11
7. Gibson assembly 11
8. 勝任細胞(Competent cell)製備 12
9. 熱休克轉形作用(Heat-shock transformation ) 12
10. 質體快速篩選 (Quick screening) 12
11. 接合生殖 (Conjugation) 13
12. 無標記基因突變法 (Marker-less mutagenesis) 13
13. 菌落聚合酶鏈鎖反應 (Colony PCR) 14
14. Phos-tagTM蛋白質樣本製備 14
15. Phos-tagTM蛋白質膠體電泳 (Phos-tag™ acrylamide gel analysis) 15
16. 西方墨點法 15
17. 鮑氏不動桿菌生長曲線 16
18. 最小抑菌濃度測試 (Minimal inhibitory concentration test) 17
19. 紙錠擴散測試 (Disk diffusion test) 17
20. 生物膜形成 (Biofilm formation) 18
21. 鮑氏不動桿菌感染大蠟蛾測試 18
22. RNA萃取 (RNA extraction) 19
23. 反轉錄作用 (Reverse transcription, RT) 19
24. 即時定量聚合酶鏈鎖反應 (Quantitative real-time PCR, qRT-PCR) 20
25. 轉錄體分析的樣本置備 20
結果 21
一、鮑氏不動桿菌的DJ41_1406以及DJ41_1407 21
1. DJ41_1406是雙分子調控系統調控蛋白質，命名為GacA 21
2. GacA蛋白質功能區域分析 21
3. DJ41_1407是雙分子調控系統感應蛋白質，命名為ZraS 22
4. ZraS蛋白質功能區域分析 22
5. gacA與zraS位於同一條mRNA 22
6. 鮑氏不動桿菌中的基因排列 23
二、ZraS/GacA在鮑氏不動桿菌的基礎代謝以及對抗環境壓力扮演重要角
色 24
1. 建構ΔzraS、ΔgacA以及ΔzraSΔgacA菌株 24
2. GacA影響鮑氏不動桿菌的生長 24
3. GacA影響鮑氏不動桿菌的醇類代謝 24
4. GacA影響鮑氏不動桿菌抵抗抗生素的能力 25
5. GacA影響鮑氏不動桿菌抵抗氧化壓力 26
6. GacA影響鮑氏不動桿菌生物膜形成 27
7. GacA影響鮑氏不動桿菌毒力 27
三、ZraS/GacA為一對雙分子調控系統，磷酸化GacA活化ZraS/GacA基因
轉錄 28
1. 在環境壓力或者營養源缺乏的環境下ZraS磷酸化GacA 28
2. 磷酸化GacA活化gacA和zraS基因表現 30
四、野生株、ΔzraS以及ΔgacA的轉錄體 (Transcriptome)分析 30
1. ΔzraS以及ΔgacA轉錄體的基因功能 (Gene ontology, GO)分類 30
2. GacA調控細菌的醇類、IAA以及胺基酸代謝 31
3. GacA活化醇類代謝、IAA代謝以及苯乙酸代謝的基因表現 33
五、建構pACYC184/TCSR報導基因系統 34
討論 35
一、如何研究雙分子調控系統 35
二、ZraS/GacA並非典型雙分子調控系統，GacA能夠接收來自不同感應蛋白
質的磷酸根 36
1. 在其他細菌中GacA如何調控下游基因 37
2. ZraS在其他細菌中的調控機制 38
3. 其他細菌中雙分子調控系統的交互訊息傳遞(cross-talk) 39
三、ZraS/GacA調控鮑氏不動桿菌的胺基酸、醇類以及IAA代謝 40
表一、本篇論文使用的質體 42
表二、本篇論文使用的菌種 43
表三、本篇論文使用的引子 45
表四、ZraS正調控的基因列表 48
表五、ZraS負調控的基因列表 49
表六、GacA正調控的基因列表 50
表七、GacA負調控的基因列表 59
圖一、雙分子調控系統調控蛋白質DJ41_1406演化樹 63
圖二、GacA蛋白質功能區域 64
圖三、雙分子調控系統感應蛋白質DJ41_1407演化樹 65
圖四、ZraS蛋白質功能區域 67
圖五、確認gacA與zraS位於同一條mRNA 68
圖六、鮑氏不動桿菌、克雷伯氏肺炎菌以及小束噬脯氨酸菌的基因排列示意
圖 69
圖七、確認ΔzraS、ΔgacA以及ΔgacAΔzraS菌株 70
圖八、鮑氏不動桿菌野生株、ΔzraS、ΔgacA以及ΔzraSΔgacA培養在LB培養
液中的生長曲線 72
圖九、鮑氏不動桿菌野生株、ΔzraS、ΔgacA以及ΔzraSΔgacA培養在以1%乙
醇為單一碳源的生長曲線 73
圖十、鮑氏不動桿菌野生株、ΔzraS、ΔgacA以及ΔzraSΔgacA培養在以1%
1-丙醇為單一碳源的生長曲線 74
圖十一、鮑氏不動桿菌野生株、ΔzraS、ΔgacA以及ΔzraSΔgacA培養在以
0.5%1-丁醇為單一碳源的生長曲線 75
圖十二、即時定量聚合酶鏈鎖反應分析野生株、ΔzraS、ΔgacA以及ΔzraSΔgacA
在含有1%乙醇的LB培養液中adh4基因表現量 76
圖十三、鮑氏不動桿菌野生株、ΔzraS、ΔgacA以及ΔzraSΔgacA培養在不同
抗生素中的最小抑菌濃度 77
圖十四、即時定量聚合酶鏈鎖反應分析鮑氏不動桿菌野生株在含有抗生素的環
境中gacA以及zraS的基因表現量 78
圖十五、紙錠擴散測試鮑氏不動桿菌野生株、ΔzraS、ΔgacA以及ΔzraSΔgacA
對於H2O2的抵抗能力 79
圖十六、紙錠擴散測試鮑氏不動桿菌野生株、ΔzraS、ΔgacA以及ΔzraSΔgacA
對於tBHP的抵抗能力 80
圖十七、即時定量聚合酶鏈鎖反應分析鮑氏不動桿菌野生株、ΔzraS、ΔgacA
以及ΔzraSΔgacA中ohr基因表現量 81
圖十八、測試鮑氏不動桿菌野生株、ΔzraS、ΔgacA以及ΔzraSΔgacA的生物
模形成能力 82
圖十九、鮑氏不動桿菌野生株、ΔzraS、ΔgacA以及ΔzraSΔgacA感染後大蠟
蛾幼蟲的存活率 83
圖二十、鮑氏不動桿菌野生株、ΔzraS、ΔgacA以及ΔzraSΔgacA感染後大蠟
蛾幼蟲的黑化程度 84
圖二十一、使用phos-tag™西方墨點法分析GacA磷酸化情形 85
圖二十二、使用phos-tag™西方墨點法分析磷酸化以及未磷酸化GacA 86
圖二十三、即時定量聚合酶鏈鎖反應分析野生株以及ΔzraS在不同培養狀況下
gacA基因表現量 87
圖二十四、以Gene ontology將野生株和ΔzraS轉錄體分析中基因表現量不同
的基因分類 88
圖二十五、以Gene ontology將野生株和ΔgacA轉錄體分析中基因表現量不同
的基因分類 90
圖二十六、野生株和ΔgacA的轉錄體分析中基因表現量差異大於2倍之分析
圖 92
圖二十七、野生株和ΔgacA的轉錄體分析結果中基因叢 DJ41_2041-DJ4_2054
的基因表現量差異分析圖 93
圖二十八、野生株和ΔgacA的轉錄體分析結果中基因叢 DJ41_332-DJ4_338、
DJ41_2892-DJ4_2896以及DJ41_3071-DJ4_3075的基因表現量差異分析圖
94
圖二十九、即時定量聚合酶鏈鎖反應分析野生株、ΔzraS、ΔgacA以及
ΔzraSΔgacA中adh4、ipdC、iacH以及paa基因表現量 95
圖三十、預測GacA可能結合的啟動子區域 96
圖三十一、雙分子調控系統ZraS/GacA調控的細菌基礎代謝路徑圖 97
圖三十二、pACYC184/TCSR報導基因系統示意圖 99
圖三十三、建構pACYC184/TCSR示意圖 100
圖三十四、建構pACYC184/TCSR確認圖 101
附錄一、pK18mobsacB示意圖 102
附錄二、pACYC184示意圖 103
參考文獻 104

1.Brown ED, Wright GD: Antibacterial drug discovery in the resistance era. Nature 2016, 529:336.
2.Tommasi R, Brown DG, Walkup GK, Manchester JI, Miller AA: ESKAPEing the labyrinth of antibacterial discovery. Nature Reviews Drug Discovery 2015, 14:529.
3.Antunes LCS, Visca P, Towner KJ: Acinetobacter baumannii: evolution of a global pathogen. Pathogens and Disease 2014, 71(3):292-301.
4.McConnell MJ, Actis L, Pachón J: Acinetobacter baumannii: human infections, factors contributing to pathogenesis and animal models. FEMS Microbiology Reviews 2013, 37(2):130-155.
5.Evans BA, Hamouda A, Amyes SG: The rise of carbapenem-resistant Acinetobacter baumannii. Current pharmaceutical design 2013, 19(2):223-238.
6.Nowak P, Paluchowska P: Acinetobacter baumannii: biology and drug resistance - role of carbapenemases. Folia histochemica et cytobiologica / Polish Academy of Sciences, Polish Histochemical and Cytochemical Society 2016.
7.Houang ET, Chu YW, Leung CM, Chu KY, Berlau J, Ng KC, Cheng AF: Epidemiology and infection control implications of Acinetobacter spp. in Hong Kong. Journal of clinical microbiology 2001, 39(1):228-234.
8.Baumann P: Isolation of Acinetobacter from soil and water. Journal of bacteriology 1968, 96(1):39-42.
9.Jawad A, Heritage J, Snelling AM, Gascoyne-Binzi DM, Hawkey PM: Influence of relative humidity and suspending menstrua on survival of Acinetobacter spp. on dry surfaces. Journal of clinical microbiology 1996, 34(12):2881-2887.
10.Lastoria LC, Caldeira SM, Moreira RG, Akazawa RT, Maion JC, Fortaleza CM: Ecological competition and the incidence of Acinetobacter baumannii bloodstream infections in a teaching hospital in Southeastern Brazil. Revista da Sociedade Brasileira de Medicina Tropical 2014, 47(5):583-588.
11.Li H, Liu F, Zhang Y, Wang X, Zhao C, Chen H, Zhang F, Zhu B, Hu Y, Wang H: Evolution of carbapenem-resistant Acinetobacter baumannii through whole genome sequencing and comparative genomic analysis. Antimicrobial agents and chemotherapy 2014.
12.Dijkshoorn L, Nemec A, Seifert H: An increasing threat in hospitals: multidrug-resistant Acinetobacter baumannii. Nature Reviews Microbiology 2007, 5:939.
13.Vila J, Pachón J: Acinetobacter baumannii resistant to everything: what should we do? Clinical Microbiology and Infection 2011, 17(7):955-956.
14.Kröger C, Kary SC, Schauer K, Cameron ADS: Genetic regulation of virulence and antibiotic resistance in Acinetobacter baumannii. Genes 2017, 8(1):12.
15.Stock AM, Robinson VL, Goudreau PN: Two-component signal transduction. Annual Review of Biochemistry 2000, 69(1):183-215.
16.Stranava M, Martinek V, Man P, Fojtikova V, Kavan D, Vanek O, Shimizu T, Martinkova M: Structural characterization of the heme-based oxygen sensor, AfGcHK, its interactions with the cognate response regulator, and their combined mechanism of action in a bacterial two-component signaling system. Proteins 2016, 84(10):1375-89.
17.López-Redondo ML, Moronta F, Salinas P, Espinosa J, Cantos R, Dixon R, Marina A, Contreras A: Environmental control of phosphorylation pathways in a branched two-component system. Molecular Microbiology 2010, 78(2):475-489.
18.Laub MT, Goulian M: Specificity in two-component signal transduction pathways. Annual Review of Genetics 2007, 41(1):121-145.
19.Nixon BT, Ronson CW, Ausubel FM: Two-component regulatory systems responsive to environmental stimuli share strongly conserved domains with the nitrogen assimilation regulatory genes ntrB and ntrC. Proceedings of the National Academy of Sciences of the United States of America 1986, 83(20):7850-7854.
20.Wuichet K, Cantwell BJ, Zhulin IB: Evolution and phyletic distribution of two-component signal transduction systems. Current Opinion in Microbiology 2010, 13(2):219-225.
21.Ashby MK: Survey of the number of two-component response regulator genes in the complete and annotated genome sequences of prokaryotes. FEMS Microbiology Letters 2004, 231(2):277-281.
22.Lin M-F, Lin Y-Y, Lan C-Y: The role of the two-component system baesr in disposing chemicals through regulating transporter systems in Acinetobacter baumannii. PloS one 2015, 10(7):e0132843.
23.Kröger C, Kary CS, Schauer K, Cameron DSA: Genetic regulation of virulence and antibiotic resistance in Acinetobacter baumannii. Genes 2017, 8(1).
24.De Silva PM, Kumar A: Signal transduction proteins in Acinetobacter baumannii: Role in antibiotic resistance, virulence, and potential as drug targets. Frontiers in microbiology 2019, 10:49.
25.Marchand I, Damier-Piolle L, Courvalin P, Lambert T: Expression of the RND-type efflux pump AdeABC in Acinetobacter baumannii; Is regulated by the AdeRS two-component system. Antimicrobial agents and chemotherapy 2004, 48(9):3298.
26.Ni W, Han Y, Zhao J, Wei C, Cui J, Wang R, Liu Y: Tigecycline treatment experience against multidrug-resistant Acinetobacter baumannii infections: a systematic review and meta-analysis. International Journal of Antimicrobial Agents 2016, 47(2):107-116.
27.Richmond GE, Evans LP, Anderson MJ, Wand ME, Bonney LC, Ivens A, Chua KL, Webber MA, Sutton JM, Peterson ML et al: The Acinetobacter baumannii two-component system AdeRS regulates genes required for multidrug efflux, biofilm formation, and virulence in a strain-specific manner. mBio 2016, 7(2):e00430-00416.
28.Leblanc SKD, Oates CW, Raivio TL: Characterization of the induction and cellular role of the BaeSR two-component envelope stress response of Escherichia coli. Journal of bacteriology 2011, 193(13):3367.
29.Lin M-F, Lin Y-Y, Yeh H-W, Lan C-Y: Role of the BaeSR two-component system in the regulation of Acinetobacter baumannii adeAB genes and its correlation with tigecycline susceptibility. BMC Microbiology 2014, 14(1):119.
30.Kim TY, Cha S-H, Cho S, Park Y: Tannic acid-mediated green synthesis of antibacterial silver nanoparticles. Archives of Pharmacal Research 2016, 39(4):465-473.
31.Hupkens P, Boxma H, Dokter J: Tannic acid as a topical agent in burns: historical considerations and implications for new developments. Burns 1995, 21(1):57-61.
32.Chusri S, Villanueva I, Voravuthikunchai SP, Davies J: Enhancing antibiotic activity: a strategy to control Acinetobacter infections. Journal of Antimicrobial Chemotherapy 2009, 64(6):1203-1211.
33.Jiménez-Guerra G, Heras-Cañas V, Gutiérrez-Soto M, del Pilar Aznarte-Padial M, Expósito-Ruiz M, Navarro-Marí JM, Gutiérrez-Fernández J: Urinary tract infection by Acinetobacter baumannii and Pseudomonas aeruginosa: evolution of antimicrobial resistance and therapeutic alternatives. Journal of Medical Microbiology 2018, 67(6):790-797.
34.Rolain J-M, Diene SM, Kempf M, Gimenez G, Robert C, Raoult D: Real-time sequencing to decipher the molecular mechanism of resistance of a clinical pan-drug-resistant Acinetobacter baumannii isolate from Marseille, France. Antimicrobial agents and chemotherapy 2013, 57(1):592.
35.Da Silva JG, Domingues S: Interplay between colistin resistance, virulence and fitness in Acinetobacter baumannii. Antibiotics 2017, 6(4).
36.Tomaras AP, Flagler MJ, Dorsey CW, Gaddy JA, Actis LA: Characterization of a two-component regulatory system from Acinetobacter baumannii that controls biofilm formation and cellular morphology. Microbiology (Reading, England) 2008, 154(11):3398-3409.
37.Pakharukova N, Tuittila M, Paavilainen S, Malmi H, Parilova O, Teneberg S, Knight SD, Zavialov AV: Structural basis for Acinetobacter baumannii biofilm formation. Proceedings of the National Academy of Sciences 2018, 115(21):5558.
38.Geisinger E, Mortman NJ, Vargas-Cuebas G, Tai AK, Isberg RR: A global regulatory system links virulence and antibiotic resistance to envelope homeostasis in Acinetobacter baumannii. PLoS pathogens 2018, 14(5):e1007030.
39.Geisinger E, Isberg RR: Antibiotic modulation of capsular exopolysaccharide and virulence in Acinetobacter baumannii. PLoS pathogens 2015, 11(2):e1004691.
40.Chen R, Lv R, Xiao L, Wang M, Du Z, Tan Y, Cui Y, Yan Y, Luo Y, Yang R et al: A1S_2811, a CheA/Y-like hybrid two-component regulator from Acinetobacter baumannii ATCC17978, is involved in surface motility and biofilm formation in this bacterium. MicrobiologyOpen 2017, 6(5):e00510.
41.Li J, Swanson RV, Simon MI, Weis RM: The response regulators CheB and CheY exhibit competitive binding to the kinase CheA. Biochemistry 1995, 34(45):14626-14636.
42.Cerqueira GM, Kostoulias X, Khoo C, Aibinu I, Qu Y, Traven A, Peleg AY: A global virulence regulator in Acinetobacter baumannii and its control of the phenylacetic acid catabolic pathway. The Journal of infectious diseases 2014, 210(1):46-55.
43.Tomaras AP, Dorsey CW, Edelmann RE, Actis LA: Attachment to and biofilm formation on abiotic surfaces by Acinetobacter baumannii: involvement of a novel chaperone-usher pili assembly system. Microbiology (Reading, England) 2003, 149(12):3473-3484.
44.Chen X, Kohl TA, Rückert C, Rodionov DA, Li L-H, Ding J-Y, Kalinowski J, Liu S-J: Phenylacetic acid catabolism and its transcriptional regulation in Corynebacterium glutamicum. Applied and environmental microbiology 2012, 78(16):5796.
45.Bertani G: Lysogeny at mid-twentieth century: P1, P2, and other experimental systems. Journal of bacteriology 2004, 186(3):595.
46.Saito H, Miura K-I: Preparation of transforming deoxyribonucleic acid by phenol treatment. Biochimica et Biophysica Acta (BBA) - Specialized Section on Nucleic Acids and Related Subjects 1963, 72:619-629.
47.Saiki RK, Gelfand DH, Stoffel S, Scharf SJ, Higuchi R, Horn GT, Mullis KB, Erlich HA: Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science 1988, 239(4839):487.
48.Birnboim HC, Doly J: A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic acids research 1979, 7(6):1513-1523.
49.Rhodes D, Klug A: Helical periodicity of DNA determined by enzyme digestion. Nature 1980, 286(5773):573-578.
50.Lehman IR: DNA ligase: structure, mechanism, and function. Science (New York, NY) 1974, 186(4166):790-797.
51.Gibson DG, Young L, Chuang R-Y, Venter JC, Hutchison Iii CA, Smith HO: Enzymatic assembly of DNA molecules up to several hundred kilobases. Nature methods 2009, 6:343.
52.Cohen SN, Chang AC, Boyer HW, Helling RB: Construction of biologically functional bacterial plasmids in vitro. Proceedings of the National Academy of Sciences of the United States of America 1973, 70(11):3240-3244.
53.Kado CI, Liu ST: Rapid procedure for detection and isolation of large and small plasmids. Journal of bacteriology 1981, 145(3):1365-1373.
54.Willetts N, Wilkins B: Processing of plasmid DNA during bacterial conjugation. Microbiological reviews 1984, 48(1):24-41.
55.Schäfer A, Tauch A, Jäger W, Kalinowski J, Thierbach G, Pühler A: Small mobilizable multi-purpose cloning vectors derived from the Escherichia coli plasmids pK18 and pK19: selection of defined deletions in the chromosome of Corynebacterium glutamicum. Gene 1994, 145(1):69-73.
56.J K: DNA Sequencing II : optimizing preparation and cleanup. Jones and Bartlett Publishers 2006.
57.Barbieri CM, Stock AM: Universally applicable methods for monitoring response regulator aspartate phosphorylation both in vitro and in vivo using Phos-tag-based reagents. Analytical biochemistry 2008, 376(1):73-82.
58.Kinoshita E, Takahashi M, Takeda H, Shiro M, Koike T: Recognition of phosphate monoester dianion by an alkoxide-bridged dinuclear zinc(ii) complex. Dalton Transactions 2004(8):1189-1193.
59.Renart J, Reiser J, Stark GR: Transfer of proteins from gels to diazobenzyloxymethyl-paper and detection with antisera: a method for studying antibody specificity and antigen structure. Proceedings of the National Academy of Sciences of the United States of America 1979, 76(7):3116-3120.
60.Shapiro AL, Viñuela E, V. Maizel J: Molecular weight estimation of polypeptide chains by electrophoresis in SDS-polyacrylamide gels. Biochemical and biophysical research communications 1967, 28(5):815-820.
61.Zwietering MH, Jongenburger I, Rombouts FM, van, t Riet K: Modeling of the bacterial growth curve. Applied and environmental microbiology 1990, 56(6):1875.
62.Washington JA, Yu PK: Regression curve analysis of cephalosporin activity. Appl Microbiol 1970, 19(4):589-593.
63.Niou Y-K, Wu W-L, Lin L-C, Yu M-S, Shu H-Y, Yang H-H, Lin G-H: Role of galE on biofilm formation by Thermus spp. Biochemical and biophysical research communications 2009, 390(2):313-318.
64.Hornsey M, Wareham DW: In vivo efficacy of glycopeptide-colistin combination therapies in a Galleria mellonella model of Acinetobacter baumannii Infection. Antimicrobial agents and chemotherapy 2011, 55(7):3534.
65.Dolan A, Burgess CM, Barry TB, Fanning S, Duffy G: A novel quantitative reverse-transcription PCR (qRT-PCR) for the enumeration of total bacteria, using meat micro-flora as a model. J Microbiol Methods 2009, 77(1):1-7.
66.Lindebro MC, Poellinger L, Whitelaw ML: Protein-protein interaction via PAS domains: role of the PAS domain in positive and negative regulation of the bHLH/PAS dioxin receptor-Arnt transcription factor complex. The EMBO Journal 1995, 14(14):3528-3539.
67.謝明娟: 鮑氏不動桿菌含鐵醇脫氫酶在醇類代謝及抵抗壓力中扮演的角色. 碩士論文，慈濟大學微生物及免疫學碩士班 2019.
68.Fontenelle C, Blanco C, Arrieta M, Dufour V, Trautwetter A: Resistance to organic hydroperoxides requires ohr and ohrR genes in Sinorhizobium meliloti. BMC Microbiology 2011, 11(1):100.
69.Nosova T, Jokelainen K, Kaihovaara P, Jousimies-Somer H, Siitonen A, Heine R, Salaspuro M: Aldehyde dehydrogenase activity and acetate production by aerobic bacteria representing the normal flora of human large intestine. Alcohol and alcoholism (Oxford, Oxfordshire) 1996, 31(6):555-564.
70.Lin HR, Shu HY, Lin GH: Biological roles of indole-3-acetic acid in Acinetobacter baumannii. Microbiological research 2018, 216:30-39.
71.Teufel R, Mascaraque V, Ismail W, Voss M, Perera J, Eisenreich W, Haehnel W, Fuchs G: Bacterial phenylalanine and phenylacetate catabolic pathway revealed. Proceedings of the National Academy of Sciences 2010, 107(32):14390.
72.Nwugo CC, Arivett BA, Zimbler DL, Gaddy JA, Richards AM, Actis LA: Effect of ethanol on differential protein production and expression of potential virulence functions in the opportunistic pathogen Acinetobacter baumannii. PloS one 2012, 7(12):e51936.
73.Hess JF, Bourret RB, Simon MI: Phosphorylation assays for proteins of the two-component regulatory system controlling chemotaxis in Escherichia coli. Methods in enzymology 1991, 200:188-204.
74.Scharf BE: Summary of useful methods for two-component system research. Current opinion in microbiology 2010, 13(2):246-252.
75.Sourjik V, Berg HC: Receptor sensitivity in bacterial chemotaxis. Proceedings of the National Academy of Sciences of the United States of America 2002, 99(1):123-127.
76.Kay E, Humair B, Dénervaud V, Riedel K, Spahr S, Eberl L, Valverde C, Haas D: Two GacA-dependent small RNAs modulate the quorum-sensing response in Pseudomonas aeruginosa. Journal of bacteriology 2006, 188(16):6026.
77.Kay E, Dubuis C, Haas D: Three small RNAs jointly ensure secondary metabolism and biocontrol in Pseudomonas fluorescens CHA0. Proceedings of the National Academy of Sciences of the United States of America 2005, 102(47):17136.
78.Heeb S, Haas D: Regulatory roles of the GacS/GacA two-component system in plant-associated and other gram-negative bacteria. Molecular Plant-Microbe Interactions 2001, 14(12):1351-1363.
79.Humair B, Wackwitz B, Haas D: GacA-controlled activation of promoters for small RNA genes in Pseudomonas fluorescens. Applied and environmental microbiology 2010, 76(5):1497.
80.Lapouge K, Schubert M, Allain FH-T, Haas D: Gac/Rsm signal transduction pathway of γ-proteobacteria: from RNA recognition to regulation of social behaviour. Molecular Microbiology 2008, 67(2):241-253.
81.Valverde C, Heeb S, Keel C, Haas D: RsmY, a small regulatory RNA, is required in concert with RsmZ for GacA-dependent expression of biocontrol traits in Pseudomonas fluorescens CHA0. Molecular Microbiology 2003, 50(4):1361-1379.
82.Pessi G, Williams F, Hindle Z, Heurlier K, Holden MTG, Cámara M, Haas D, Williams P: The global posttranscriptional regulator RsmA modulates production of virulence determinants and N-acylhomoserine lactones in Pseudomonas aeruginosa. Journal of bacteriology 2001, 183(22):6676.
83.Reimmann C, Beyeler M, Latifi A, Winteler H, Foglino M, Lazdunski A, Haas D: The global activator GacA of Pseudomonas aeruginosa PAO positively controls the production of the autoinducer N-butyryl-homoserine lactone and the formation of the virulence factors pyocyanin, cyanide, and lipase. Molecular Microbiology 1997, 24(2):309-319.
84.Cha JY, Lee DG, Lee JS, Oh JI, Baik HS: GacA directly regulates expression of several virulence genes in Pseudomonas syringae pv. tabaci 11528. Biochemical and biophysical research communications 2012, 417(2):665-672.
85.Vogel J, Wagner EGH: Target identification of small noncoding RNAs in bacteria. Current opinion in microbiology 2007, 10(3):262-270.
86.Petit-Hartlein I, Rome K, de Rosny E, Molton F, Duboc C, Gueguen E, Rodrigue A, Coves J: Biophysical and physiological characterization of ZraP from Escherichia coli, the periplasmic accessory protein of the atypical ZraSR two-component system. Biochem J 2015, 472(2):205-216.
87.Rome K, Borde C, Taher R, Cayron J, Lesterlin C, Gueguen E, De Rosny E, Rodrigue A: The two-component system ZraPSR is a novel ESR that contributes to intrinsic antibiotic tolerance in Escherichia coli. J Mol Biol 2018, 430(24):4971-4985.
88.Lee LJ, Barrett JA, Poole RK: Genome-wide transcriptional response of chemostat-cultured Escherichia coli to zinc. Journal of bacteriology 2005, 187(3):1124-1134.
89.Appia-Ayme C, Hall A, Patrick E, Rajadurai S, Clarke TA, Rowley G: ZraP is a periplasmic molecular chaperone and a repressor of the zinc-responsive two-component regulator ZraSR. Biochem J 2012, 442(1):85-93.
90.Tschauner K, Hornschemeyer P, Muller VS, Hunke S: Dynamic interaction between the CpxA sensor kinase and the periplasmic accessory protein CpxP mediates signal recognition in E. coli. PloS one 2014, 9(9):e107383.
91.Wei J-R, Tsai Y-H, Soo P-C, Horng Y-T, Hsieh S-C, Ho S-W, Lai H-C: Biochemical characterization of RssA-RssB, a two-component signal transduction system regulating swarming behavior in Serratia marcescens. Journal of bacteriology 2005, 187(16):5683.
92.Soo P-C, Horng Y-T, Wei J-R, Shu J-C, Lu C-C, Lai H-C: Regulation of swarming motility and flhDCSm expression by RssAB signaling in Serratia marcescens. Journal of bacteriology 2008, 190(7):2496.
93.Wei CF, Tsai YH, Tsai SH, Lin CS, Chang CJ, Lu CC, Huang HC, Lai HC: Cross-talk between bacterial two-component systems drives stepwise regulation of flagellar biosynthesis in swarming development. Biochemical and biophysical research communications 2017, 489(1):70-75.
94.Clarke MB, Sperandio V: Transcriptional autoregulation by quorum sensing Escherichia coli regulators B and C (QseBC) in enterohaemorrhagic E. coli (EHEC). Molecular Microbiology 2005, 58(2):441-455.
95.Guckes KR, Kostakioti M, Breland EJ, Gu AP, Shaffer CL, Martinez CR, Hultgren SJ, Hadjifrangiskou M: Strong cross-system interactions drive the activation of the QseB response regulator in the absence of its cognate sensor. Proceedings of the National Academy of Sciences 2013, 110(41):16592.
96.Guckes KR, Breland EJ, Zhang EW, Hanks SC, Gill NK, Algood HM, Schmitz JE, Stratton CW, Hadjifrangiskou M: Signaling by two-component system noncognate partners promotes intrinsic tolerance to polymyxin B in uropathogenic Escherichia coli. Sci Signal 2017, 10(461).
97.Urano H, Yoshida M, Ogawa A, Yamamoto K, Ishihama A, Ogasawara H: Cross-regulation between two common ancestral response regulators, HprR and CusR, in Escherichia coli. Microbiology (Reading, England) 2017, 163(2):243-252.
98.Lee Y, Lee DH, Kho CW, Lee AY, Jang M, Cho S, Lee CH, Lee JS, Myung PK, Park BC et al: Transthyretin-related proteins function to facilitate the hydrolysis of 5-hydroxyisourate, the end product of the uricase reaction. FEBS letters 2005, 579(21):4769-4774.
99.Munson GP, Lam DL, Outten FW, Halloran TV: Identification of a copper-responsive two-component system on the chromosome of Escherichia coli K-12. Journal of bacteriology 2000, 182(20):5864.
100.Lin H-R, Shu H-Y, Lin G-H: Biological roles of indole-3-acetic acid in Acinetobacter baumannii. Microbiological research 2018, 216:30-39.
101.Chang AC, Cohen SN: Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid. Journal of bacteriology 1978, 134(3):1141.
102.de Lorenzo V, Fernández S, Herrero M, Jakubzik U, Timmis KN: Engineering of alkyl- and haloaromatic-responsive gene expression with mini-transposons containing regulated promoters of biodegradative pathways of Pseudomonas. Gene 1993, 130(1):41-46.
103.Bouvet PJM, Grimont PAD: Taxonomy of the genus Acinetobacter with the recognition of Acinetobacter baumannii sp. nov., Acinetobacter haemolyticus sp. nov., Acinetobacter johnsonii sp. nov., and Acinetobacter junii sp. nov. and emended descriptions of Acinetobacter calcoaceticus and Acinetobacter lwoffii. International Journal of Systematic and Evolutionary Microbiology 1986, 36(2):228-240.