The RhlR quorum-sensing receptor controls Pseudomonas aeruginosa pathogenesis and biofilm development independently of its canonical homoserine lactone autoinducer

PLoS Pathogens

Volumn 13, Issue 7, 2017, Pages

  Mukherjee, Sampriti ^a   Moustafa, Dina ^b   Smith, Chari D ^a   Goldberg, Joanna B ^b   Bassler, Bonnie L ^a,c  

^a PRINCETON UNIVERSITY   (United States)

^b EMORY UNIVERSITY SCHOOL OF MEDICINE   (United States)

^c HOWARD HUGHES MEDICAL INSTITUTE   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

N (3 OXODODECANOYL)HOMOSERINE LACTONE; PYOCYANINE; VIRULENCE FACTOR; BACTERIAL PROTEIN; GAMMA BUTYROLACTONE; HOMOSERINE LACTONE; RHLR PROTEIN, PSEUDOMONAS AERUGINOSA;

ARTICLE; BACTERICIDAL ACTIVITY; BIOFILM; CONTROLLED STUDY; DNA TRANSCRIPTION; GENE DELETION; GENE EXPRESSION; GENE EXPRESSION REGULATION; GENE MUTATION; LIQUID CHROMATOGRAPHY-MASS SPECTROMETRY; NONHUMAN; PHENOTYPE; PSEUDOMONAS AERUGINOSA; PSEUDOMONAS INFECTION; QUORUM SENSING; REVERSE TRANSCRIPTION POLYMERASE CHAIN REACTION; RNA SEQUENCE; SIGNAL TRANSDUCTION; TRANSCRIPTION INITIATION; ANALOGS AND DERIVATIVES; ANIMAL; BAGG ALBINO MOUSE; FEMALE; GENETICS; HUMAN; METABOLISM; MICROBIOLOGY; MOUSE; PATHOGENICITY; PHYSIOLOGY; REGULON; VIRULENCE;

4-BUTYROLACTONE; ANIMALS; BACTERIAL PROTEINS; BIOFILMS; FEMALE; GENE EXPRESSION REGULATION, BACTERIAL; HUMANS; MICE; MICE, INBRED BALB C; PSEUDOMONAS AERUGINOSA; PSEUDOMONAS INFECTIONS; QUORUM SENSING; REGULON; VIRULENCE;

EID: 85026878471     PISSN: 15537366     EISSN: 15537374     Source Type: Journal    
DOI: 10.1371/journal.ppat.1006504     Document Type: Article

References (88)

1. Miller MB, Bassler BL, QUORUM SENSING IN BACTERIA. Annu. Rev. Microbiol.2001. 55: 165–99. doi: 10.1146/annurev.micro.55.1.165 11544353
2. Lazdunski AM, Ventre I, Sturgis JN, Regulatory circuits and communication in Gram-negative bacteria. Nat. Rev. Microbiol.2004. 2: 581–592. doi: 10.1038/nrmicro924 15197393
3. Waters CM, Bassler BL, QUORUM SENSING: Cell-to-Cell Communication in Bacteria. Annu. Rev. Cell Dev. Biol.2005. 21: 319–346. doi: 10.1146/annurev.cellbio.21.012704.131001 16212498
4. Papenfort K, Bassler BL, Quorum sensing signal-response systems in Gram-negative bacteria. Nat. Rev. Microbiol.2016. 14: 576–88. doi: 10.1038/nrmicro.2016.89 27510864
5. Fuqua C, Greenberg EP, Listening in on bacteria: acyl-homoserine lactone signalling. Nat. Rev. Mol. cell Biol.2002. 3: 685–695. doi: 10.1038/nrm907 12209128
6. Zhang R, Pappas KM, Brace JL, Miller PC, Oulmassov T, Molyneaux JM, et al. Structure of a bacterial quorum-sensing transcription factor complexed with pheromone and DNA. Nature2002.417: 971–974. doi: 10.1038/nature00833 12087407
7. Churchill MEA, Chen L, Structural basis of acyl-homoserine lactone-dependent signaling. Chem. Rev.2011. 111: 68–85. doi: 10.1021/cr1000817 21125993
8. Urbanowski ML, Lostroh CP, Greenberg EP, Reversible Acyl-Homoserine Lactone Binding to Purified Vibrio fischeri LuxR Protein. J. Bacteriol.2004. 186: 631–637. doi: 10.1128/JB.186.3.631-637.2004 14729687
9. Weingart CL, White CE, Liu S, Chai Y, Cho H, Tsai CS, Wei Y, Delay NR, Gronquist M.R., Eberhard A., et al. (2005). Direct binding of the quorum sensing regulator CepR of Burkholderia cenocepacia to two target promoters in vitro. Mol. Microbiol.57, 452–467. doi: 10.1111/j.1365-2958.2005.04656.x 15978077
10. Chai Y, Winans SC, Amino-terminal protein fusions to the TraR quorum-sensing transcription factor enhance protein stability and autoinducer-independent activity. J. Bacteriol.2005; 187: 1219–1226. doi: 10.1128/JB.187.4.1219-1226.2005 15687185
11. Rutherford ST, Bassler BL, Bacterial Quorum Sensing: Its Role in Virulence and Possibilities for Its Control. Cold Spring Harb. Perspect. Med.2012; 2:a012427. doi: 10.1101/cshperspect.a012427 23125205
12. Smith EE, Buckley DG, Wu Z, Saenphimmachak C, Hoffman LR, Miller SI, et al. Genetic adaptation by Pseudomonas aeruginosa to the airways of cystic fibrosis patients. Sci. York2006; 103: 8487–8492.
13. Rumbaugh KP, Diggle SP, Watters CM, Ross-Gillespie A, Griffin AS, West SA, Quorum Sensing and the Social Evolution of Bacterial Virulence. Curr. Biol.2009; 19: 341–345. doi: 10.1016/j.cub.2009.01.050 19230668
14. Seed PC, Passador L, Iglewski BH, Activation of the Pseudomonas aeruginosa lasI gene by LasR and the Pseudomonas autoinducer PAI: An autoinduction regulatory hierarchy. J. Bacteriol.1995; 177: 654–659. 7836299
15. Pearson JP, Passador L, Iglewski BH, Greenberg EP, A second N-acylhomoserine lactone signal produced by Pseudomonas aeruginosa. Proc. Natl. Acad. Sci. U. S. A.1995; 92: 1490–1494. 7878006
16. Brint JM, Ohman DE, Synthesis of multiple exoproducts in Pseudomonas aeruginosa is under the control of RhlR-RhlI, another set of regulators in strain PAO1 with homology to the autoinducer-responsive LuxR-LuxI family. J. Bacteriol.1995; 177: 7155–7163. 8522523
17. De Kievit TR, Kakai Y, Register JK, Pesci EC, Iglewski BH, Role of the Pseudomonas aeruginosa las and rhl quorum-sensing systems in rhlI regulation. FEMS Microbiol. Lett.2002; 212: 101–106. 12076794
18. Wagner VE, Bushnell D, Passador L, Brooks AI, Iglewski BH, Microarray Analysis of Pseudomonas aeruginosa Quorum-Sensing Regulons: Effects of Growth Phase and Environment. J. Bacteriol.2003; 185: 2080–2095. doi: 10.1128/JB.185.7.2080-2095.2003 12644477
19. Winson MK, Camarat M, Latifi A, Foglino M, Chhabrat SRAM, Daykint M, et al. Multiple N-acyl-L-homoserine lactone signal molecules regulate production of virulence determinants and secondary metabolites in Pseudomonas aeruginosa. 1995; 92: 9427–9431. 7568146
20. Whiteley M, Lee KM, Greenberg EP, Identification of genes controlled by quorum sensing in Pseudomonas aeruginosa. Proc. Natl. Acad. Sci. U. S. A.1999. 96: 13904–13909. 10570171
21. Schuster M, Greenberg EP, Early activation of quorum sensing in Pseudomonas aeruginosa reveals the architecture of a complex regulon. BMC Genomics. 2007; 8: 287. doi: 10.1186/1471-2164-8-287 17714587
22. Pearson JP, Feldman M, Iglewski BH, Prince A, Pseudomonas aeruginosa Cell-to-Cell Signaling Is Required for Virulence in a Model of Acute Pulmonary Infection. Infect. Immun.2000; 68: 4331–4334. 10858254
23. Rumbaugh KP, Griswold JA, Hamood AN, The role of quorum sensing in the in vivo virulence of Pseudomonas aeruginosa. Microbes Infect.2000; 2: 1721–1731. 11137045
24. Chen G, Swem LR, Swem DL, Stauff DL, O’Loughlin CT, Jeffrey PD, et al. A Strategy for Antagonizing Quorum Sensing. Mol. Cell. 2011; 42: 199–209. doi: 10.1016/j.molcel.2011.04.003 21504831
25. LaSarre B, Federle MJ, Exploiting Quorum Sensing To Confuse Bacterial Pathogens. Microbiol. Mol. Biol. Rev.2013; 77: 73–111. doi: 10.1128/MMBR.00046-12 23471618
26. O’Loughlin CT, Miller LC, Siryaporn A, Drescher K, Semmelhack MF, Bassler BL, A quorum-sensing inhibitor blocks Pseudomonas aeruginosa virulence and biofilm formation. Proc. Natl. Acad. Sci.2013; 110: 17981–17986. doi: 10.1073/pnas.1316981110 24143808
27. Davies DG, Parsek MR, Pearson JP, Iglewski BH, Costerton JW, Greenberg EP, The Involvement of Cell-to-Cell Signals in the Development of a Bacterial Biofilm. Science. 1998; 280: 295–298. 9535661
28. Friedman L, Kolter R, Genes involved in matrix formation in Pseudomonas aeruginosa PA14 biofilms. Mol. Microbiol.2004; 51: 675–690. 14731271
29. James GA, Swogger E, Wolcott R, Pulcini ED, Secor P, Sestrich J, et al. Biofilms in chronic wounds. Wound Repair Regen.2008; 16: 37–44. doi: 10.1111/j.1524-475X.2007.00321.x 18086294
30. Dietrich LEP, Teal TK, Price-Whelan A, Newman DK, Redox-Active Antibiotics Control Gene Expression and Community Behavior in Divergent Bacteria. Science. 2008; 321: 1203–1206. doi: 10.1126/science.1160619 18755976
31. Dietrich LEP, Okegbe C, Price-Whelan A, Sakhtah H, Hunter RC, Newman DK, Bacterial community morphogenesis is intimately linked to the intracellular redox state. J. Bacteriol.2013; 195: 1371–1380. doi: 10.1128/JB.02273-12 23292774
32. Colvin KM, Irie Y, Tart CS, Urbano R, Whitney JC, Ryder C, et al. The Pel and Psl polysaccharides provide Pseudomonas aeruginosa structural redundancy within the biofilm matrix. Environ. Microbiol.2012; 14: 1913–1928. doi: 10.1111/j.1462-2920.2011.02657.x 22176658
33. Recinos DA, Sekedat MD, Hernandez A, Cohen TS, Sakhtah H, Prince AS, et al. Redundant phenazine operons in Pseudomonas aeruginosa exhibit environment-dependent expression and differential roles in pathogenicity. Proc. Natl. Acad. Sci.2012; 109: 19420–19425. doi: 10.1073/pnas.1213901109 23129634
34. Marvig RL, Sommer LM, Molin S, Johansen HK, Convergent evolution and adaptation of Pseudomonas aeruginosa within patients with cystic fibrosis. Nat. Genet.2014; 47: 1–9.
35. LaFayette SL, Houle D, Beaudoin T, Wojewodka G, Radzioch D, Hoffman LR, et al. Cystic fibrosis-adapted Pseudomonas aeruginosa quorum sensing lasR mutants cause hyperinflammatory responses. Sci. Adv.2015; 1: 1–16.
36. Feltner JB, Wolter DJ, Pope CE, Groleau MC, Smalley NE, Greenberg EP, et al. LasR variant cystic fibrosis isolates reveal an adaptable quorum-sensing hierarchy in Pseudomonas aeruginosa. MBio. 2016; 7: 1–9.
37. Limmer S, Haller S, Drenkard E, Lee J, Yu S, Kocks C, Ausubel FM, Ferrandon D, Pseudomonas aeruginosa RhlR is required to neutralize the cellular immune response in a Drosophila melanogaster oral infection model. Proc. Natl. Acad. Sci. U. S. A.2011; 108: 17378–83. doi: 10.1073/pnas.1114907108 21987808
38. Pesci EC, Pearson JP, Seed PC, Iglewski BH, Regulation of las and rhl quorum sensing in Pseudomonas aeruginosa. J. Bacteriol.1997; 179: 3127–3132. 9150205
39. Dekimpe V, Déziel E, Revisiting the quorum-sensing hierarchy in Pseudomonas aeruginosa: The transcriptional regulator RhlR regulates LasR-specific factors. Microbiology. 2009; 155: 712–723. doi: 10.1099/mic.0.022764-0 19246742
40. Høyland-Kroghsbo NM, Paczkowski J, Mukherjee S, Broniewski J, Westra E, Bondy-Denomy J, et al. Quorum sensing controls the Pseudomonas aeruginosa CRISPR-Cas adaptive immune system. Proc. Natl. Acad. Sci. U. S. A.2017; 114: 201617415.
41. Paczkowski J, Mukherjee S, McCready AR, Cong JP, Aquino CJ, Kim H, et al. Flavonoids suppress Pseudomonas aeruginosa virulence through allosteric inhibition of quorum-sensing receptors. J. Biol. Chem.2017; 292: 4064–4076. doi: 10.1074/jbc.M116.770552 28119451
42. Monds RD, O’Toole GA, The developmental model of microbial biofilms: ten years of a paradigm up for review. Trends Microbiol. 2009; 17: 73–87. doi: 10.1016/j.tim.2008.11.001 19162483
43. Ferrara S, Brugnoli M, de Bonis A, Righetti F, Delvillani F, Dehò G, et al. Comparative profiling of Pseudomonas aeruginosa strains reveals differential expression of novel unique and conserved small RNAs. PLoS One. 2012; 7.
44. Boles B.R., Thoendel M., Singh P.K., (2005). Rhamnolipids mediate detachment of Pseudomonas aeruginosa from biofilms. Mol. Microbiol.57, 1210–1223. doi: 10.1111/j.1365-2958.2005.04743.x 16101996
45. Bhattacharjee A, Nusca TD, Hochbaum AI, Rhamnolipids mediate an interspecies biofilm dispersal signaling pathway. ACS Chem. Biol.2016; 11: 3068–3076. doi: 10.1021/acschembio.6b00750 27623227
46. Soberón-Chávez G, Aguirre-Ramírez M, Sánchez R, The Pseudomonas aeruginosa RhlA enzyme is involved in rhamnolipid and polyhydroxyalkanoate production. J. Ind. Microbiol. Biotechnol.2005; 32: 675–677. doi: 10.1007/s10295-005-0243-0 15937697
47. Reis RS, Pereira AG, Neves BC, Freire DMG, Gene regulation of rhamnolipid production in Pseudomonas aeruginosa—A review. Bioresour. Technol.2011; 102: 6377–6384. doi: 10.1016/j.biortech.2011.03.074 21498076
48. Chugani S, Greenberg EP, LuxR homolog-independent gene regulation by acyl-homoserine lactones in Pseudomonas aeruginosa. Proc. Natl. Acad. Sci. U. S. A.2010; 107: 10673–10678. doi: 10.1073/pnas.1005909107 20498077
49. Pesci EC, Milbank JBJ, Pearson JP, McKnight S, Kende AS, Greenberg EP, et al. Quinolone signaling in the cell-to-cell communication system of Pseudomonas aeruginosa. Proc. Natl. Acad. Sci.1999; 96: 11229–11234. 10500159
50. Yates EA, Philipp B, Buckley C, Atkinson S, Chhabra SR, Sockett RE, et al. N -Acylhomoserine Lactones Undergo Lactonolysis in a pH-, Temperature-, and Acyl Chain Length-Dependent Manner during Growth of Yersinia pseudotuberculosis and Pseudomonas aeruginosa. J. Bacteriol.2002; 70: 5635–5646.
51. Tan MW, Mahajan-Miklos S, Ausubel FM, Killing of Caenorhabditis elegans by Pseudomonas aeruginosa used to model mammalian bacterial pathogenesis. Proc. Natl. Acad. Sci. U. S. A.1999; 96: 715–20. 9892699
52. Lee J, Zhang L, The hierarchy quorum sensing network in Pseudomonas aeruginosa. Protein Cell. 2014; 6: 26–41. doi: 10.1007/s13238-014-0100-x 25249263
53. Reverchon S, Bouillant ML, Salmond G, Nasser W, Integration of the quorum-sensing system in the regulatory networks controlling virulence factor synthesis in Erwinia chrysanthemi. Mol. Microbiol.1998; 29: 1407–1418. 9781878
54. Minogue TD, Wehland-Von T M, Bernhard F, Von Bodman SB, The autoregulatory role of EsaR, a quorum-sensing regulator in Pantoea stewartii ssp. stewartii: Evidence for a repressor function. Mol. Microbiol.2002; 44: 1625–1635. 12067349
55. Burr T, Barnard AML, Corbett MJ, Pemberton CL, Simpson NJL, Salmond GPC, Identification of the central quorum sensing regulator of virulence in the enteric phytopathogen, Erwinia carotovora: The VirR repressor. Mol. Microbiol.2006; 59: 113–125. doi: 10.1111/j.1365-2958.2005.04939.x 16359322
56. Brachmann AO, Brameyer S, Kresovic D, Hitkova I, Kopp Y, Manske C, et al. Pyrones as bacterial signaling molecules. Nat. Chem. Biol.2013. 9: 573–8. doi: 10.1038/nchembio.1295 23851573
57. Brameyer S, Kresovic D, Bode HB, Heermann R, Dialkylresorcinols as bacterial signaling molecules. Proc. Natl. Acad. Sci.2015. 112: 572–577. doi: 10.1073/pnas.1417685112 25550519
58. Hawver LA, Jung SA, Ng W, Specificity and complexity in bacterial quorum-sensing systems. FEMS Microbiol. Rev.2016. 738–752. doi: 10.1093/femsre/fuw014 27354348
59. Chandler JR, Heilmann S, Mittler JE, Greenberg EP, Acyl-homoserine lactone-dependent eavesdropping promotes competition in a laboratory co-culture model. ISME J.2012. 6: 1–10. doi: 10.1038/ismej.2011.71
60. Kim T, Duong T, Wu CA, Choi J, Lan N, Kang SW, et al. Structural insights into the molecular mechanism of Escherichia coli SdiA, a quorum-sensing receptor. Acta Crystallogr. Sect. D Biol. Crystallogr. 2014. 70: 694–707.
61. Fuqua C, Winans SC, Greenberg EP, CENSUS AND CONSENSUS IN BACTERIAL ECOSYSTEMS: The LuxR-LuxI Family of Quorum-Sensing Transcriptional Regulators. Annu. Rev. Microbiol.1996. 50: 727–51. doi: 10.1146/annurev.micro.50.1.727 8905097
62. Grosso-Becerra MV, Croda-García G, Merino E, Servín-González L, Mojica-Espinosa R, Soberón-Chávez G, Regulation of Pseudomonas aeruginosa virulence factors by two novel RNA thermometers. Proc. Natl. Acad. Sci. U. S. A.2014. 111: 15562–7. doi: 10.1073/pnas.1402536111 25313031
63. Moghal N, Sternberg PW, Multiple positive and negative regulators of signaling by the EGF-receptor. Curr. Opin. Cell Biol.1999. 11: 190–196. 10209155
64. Wilkinson SP, Grove A, Ligand-responsive transcriptional regulation by members of the MarR family of winged helix proteins. Curr. Issues Mol. Biol.2006. 8: 51–62. 16450885
65. Cezairliyan B, Vinayavekhin N, Grenfell-Lee D, Yuen GJ, Saghatelian A, Ausubel FM, Identification of Pseudomonas aeruginosa Phenazines that Kill Caenorhabditis elegans. PLoS Pathog.2013. 9.
66. Zhu H, Bandara R, Conibear TC, Thuruthyil SJ, Rice SA, Kjelleberg S, et al. Pseudomonas aeruginosa with lasI quorum-sensing deficiency during corneal infection. Invest Ophthalmol Vis Sci. 2004. 45(6):1897–903. 15161855
67. Tang HB, Mango EDI, Bryan R, Gambello M, Iglewski BH, Goldberg JB, et al. Contribution of Specific Pseudomonas aeruginosa Virulence Factors to Pathogenesis of Pneumonia in a Neonatal Mouse Model of Infection. Infect. Immun.1996. 64: 37–43. 8557368
68. Lazenby JJ, Griffin PE, Kyd J, Whitchurch CB, Cooley MA, A Quadruple Knockout of lasIR and rhlIR of Pseudomonas aeruginosa PAO1 That Retains Wild-Type Twitching Motility Has Equivalent Infectivity and Persistence to PAO1 in a Mouse Model of Lung Infection. PloS One. 2013; 8: e60973. doi: 10.1371/journal.pone.0060973 23593362
69. Wu H, Song Z, Givskov M, Doring G, Worlitzsch D, Mathee K, Rygaard J, et al. Pseudomonas aeruginosa mutations in lasI and rhlI quorum sensing systems result in milder chronic lung infection. Microbiology. 2016; 2318: 34–1105.
70. Lau GW, Ran H, Kong F, Hassett DJ, Mavrodi D, Pseudomonas aeruginosa Pyocyanin Is Critical for Lung Infection in Mice. Infect. Immun.2004. 72: 4275–4278. doi: 10.1128/IAI.72.7.4275-4278.2004 15213173
71. Kukavica-ibrulj I, Bragonzi A, Paroni M, Winstanley C, O’Toole GA, Levesque RC, In Vivo Growth of Pseudomonas aeruginosa Strains PAO1 and PA14 and the Hypervirulent Strain LESB58 in a Rat Model of Chronic Lung Infection. J. Bacteriol.2008. 190: 2804–2813. doi: 10.1128/JB.01572-07 18083816
72. Wareham DW, Papakonstantinopoulou A, Curtis MA, The Pseudomonas aeruginosa PA14 type III secretion system is expressed but not essential to virulence in the Caenorhabditis elegans–P. aeruginosa pathogenicity model. FEMS Microbiol. Lett.2005. 242: 209–216. doi: 10.1016/j.femsle.2004.11.018 15621439
73. Mah TF, Pitts B, Pellock B, Walker GC, Stewart PS, O’Toole GA, A genetic basis for Pseudomonas aeruginosa biofilm antibiotic resistance. Lett. to Nat.2003. 426: 1–5.
74. Rice LB, Federal funding for the study of Antimicrobial Resistance in Nosocomial Pathogens: No ESKAPE. J. Infect. Dis.2008. 197:1079–81. doi: 10.1086/533452 18419525
75. Pendleton JN, Gorman SP, Gilmore BF, Clinical relevance of the ESKAPE pathogens. Expert Rev. Anti-Infective Therapy2013. 11(3): 297–308. doi: 10.1586/eri.13.12 23458769
76. Tacconelli E, Carmell Y, Harbarth S, Kahlmeter G, Kluytmans J, Mendelson M, et al. Global priority list of antibiotic-resistant bacteria to guide research, discovery, and development of new antibiotics. WHO Press Release2017.
77. Gibson DG, Young L, Chuang R, Venter JC, Iii CAH, Smith HO, et al. Enzymatic assembly of DNA molecules up to several hundred kilobases. Nat. Mathods2009. 6: 12–16.
78. Hmelo LR, Borlee BR, Almblad H, Love ME, Randall TE, Tseng BS, et al. Precision-engineering the Pseudomonas aeruginosa genome with two-step allelic exchange. Nat. Protoc.2015. 10: 1820–41. doi: 10.1038/nprot.2015.115 26492139
79. Schweizer HP, Escherichia-Pseudomonas Shuttle Vectors derived from pUC18/19. Gene1991. 97: 109–112 1899844
80. Shaner NC, Lambert GG, Chammas A, Ni Y, Cranfill PJ, Baird M, et al.A bright monomeric green fluorescent protein derived from Branchiostoma lanceolatum. Nat. Methods2013. 10: 407–409. doi: 10.1038/nmeth.2413 23524392
81. Damron FH, Mckenney ES, Schweizer HP, Goldberg B, Construction of a Broad-Host-Range Tn 7 -Based Vector for Single- Copy P BAD -Controlled Gene Expression in Gram-Negative Bacteria. 2013. 79: 718–721.
82. Choi KH, Kumar A, Schweizer HP, A 10-min method for preparation of highly electrocompetent Pseudomonas aeruginosa cells: Application for DNA fragment transfer between chromosomes and plasmid transformation. J. Microbiol. Methods2006. 64: 391–397. doi: 10.1016/j.mimet.2005.06.001 15987659
83. Damron FH, McKenney ES, Barbier M, Liechti GW, Schweizer HP, Goldberg JB, Construction of mobilizable mini-Tn7 vectors for bioluminescent detection of gram-negative bacteria and single-copy promoter lux reporter analysis. Appl. Environ. Microbiol.2013. 79: 4149–4153. doi: 10.1128/AEM.00640-13 23584769
84. Essar DW, Eberly LEE, Hadero A, Crawford IP, Identification and Characterization of Genes for a Second Anthranilate Synthase in Pseudomonas aeruginosa: Interchangeability of the Two Anthranilate Synthases and Evolutionary Implications. 1990. 884–900. 2153661
85. Winsor GL, Griffiths EJ, Lo R, Dhillon BK, Shay JA, Brinkman FS, Enhanced annotations and features for comparing thousands of Pseudomonas genomes in the Pseudomonas genome database. Nucleic Acids Res. 2016. doi: 10.1093/nar/gkv1227 26578582
86. Brenner S, The genetics of Caenorhabditis elegans. Genetics.1974. 77: 71–94. 4366476
87. Revelli DA, Boylan JA, Gherardini FC, Bowen RA, State C, A non-invasive intratracheal inoculation method for the study of pulmonary melioidosis. Front. Cell Infect. Microbiol.2012. 2: 1–8. doi: 10.3389/fcimb.2012.00001
88. Reed LJ, Muench H, A simple method of estimating fifty percent endpoints. Am J Hyg.1938. 27: 493–497.