CONSTRUCTION AND BIOLOGICAL CHARACTERISTICS ANALYSIS OF <i>CPXRA</i> MUTANT IN <i>AEROMONAS HYDROPHILA</i>

Lan-Li ZHANG; Qian-Qian ZHANG; Hui CHEN; Xie-Hao WANG; Zhen-Bing WU; Yu-Qing FENG; Ai-Hua LI

doi:10.7541/2018.001

Volume 42 Issue 1

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ACTA HYDROBIOLOGICA SINICA > 2018 > 42(1): 1-10. > DOI: 10.7541/2018.001

Lan-Li ZHANG, Qian-Qian ZHANG, Hui CHEN, Xie-Hao WANG, Zhen-Bing WU, Yu-Qing FENG, Ai-Hua LI. CONSTRUCTION AND BIOLOGICAL CHARACTERISTICS ANALYSIS OF CPXRA MUTANT IN AEROMONAS HYDROPHILA[J]. ACTA HYDROBIOLOGICA SINICA, 2018, 42(1): 1-10. DOI: 10.7541/2018.001

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CONSTRUCTION AND BIOLOGICAL CHARACTERISTICS ANALYSIS OF CPXRA MUTANT IN AEROMONAS HYDROPHILA

1.
State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China
2.
University of Chinese Academy of Sciences, Beijing 100049, China
3.
Freshwater Aquaculture Collaborative Innovation Center of Hubei Province, Wuhan 430070, China
4.
Fisheries Technology Extension Center in Jiangsu province, Nanjing 210036, China

Received Date: March 08, 2017
Rev Recd Date: June 17, 2017
Available Online: January 15, 2018
Published Date: December 31, 2017

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Abstract

To investigate the roles of two Cpx component systems in the growth and virulence of Aeromonas hydrophila, we constructed the cpxRA gene cluster mutant Δcpx with the deletion of 57—1879 bp using suicide plasmid pRE112 as the vector. This procedure was based on fusion PCR and gene homologous recombination principles. Through electrophoresis and fluorescence quantitative PCR, the partial deletion of the cpxRA gene cluster in the mutant was confirmed. Then we compared the differences between mutant and wild strains in biological characteristics including growth, biofilm formation, stress tolerance and virulence. The results showed that the mutant had no significant difference with the wild strain in morphology, growth, biofilm formation and virulence. The main differences existed in the response to high osmotic pressure, SDS (Sodium dodecyl sulfate), EDTA (Ethylene Diamine Tetraacetic Acid) and polymyxin B stimulation. This study reveals that the Cpx system of Aeromonas hydrophila is involved in the response to external stimulus factors, and plays a relatively minor role in virulence.
- Aeromonas hydrophila,
- cpxRA,
- pRE112,
- Stress tolerance

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[1]	Thune R L, Stanley L A, Cooper R K. Pathogenesis of gram-negative bacterial infections in warm water fish [J]. Annual Reviews of Fish Diseases, 1993, 3: 145—185
[2]	Popoff M. Aeromonas. Krieg N R, editor. Bergy’s Manual of Systematic Bacteriology, vol. 1. Baltimore [M]. Williams &Wilkins, 1984, 545—548
[3]	Altwegg M, Geiss H K. Aeromonas as a human pathogen [J]. Critical Reviews in Microbiology, 1989, 16: 253—286
[4]	Austin B, Austin D A. Bacterial fish pathogens. Disease of Farmed and Wild Fish [M]. 5th ed Chichester: Springer Praxis, 2012, 119—146
[5]	Cipriano R C. Aeromonas hydrophila and Motile Aeromonad Septicemias of Fish [M]. Fish Disease Leaflet, 2001, 68
[6]	Conrad M. Cross-scale information processing in evolution, development and intelligence [J]. Biosystems, 1996, 38(2—3): 97—109
[7]	Raivio T L, Silhavy T J. Transduction of envelope stress in Escherichia coli by the Cpx two-component system [J]. Journal of Bacteriology, 1997, 179(24): 7724—7733
[8]	张旭杰, 杨五名, 李彤彤, 等. 湖北地区暴发病池塘中嗜水气单胞菌的遗传多样性和毒力特征研究. 水生生物学报, 2013, 37(3): 458—466 Zhang X J, Yang W M, Li T T, et al. The genetic diversity and virulence characteristics of Aeromonas hydrophila isolated from fishponds with disease outbreaks in Hubei province [J]. Acta Hydrobiologica Sinica, 2013, 37(3): 458—466
[9]	Ho S N, Hunt H D, Horton R M, et al. Site-directed mutagenesis by overlap extension using the polymerase chain-reaction [J]. Gene, 1989, 77(1): 51—59
[10]	Hu Y H, Liu C S, Hou J H, et al. Identification, characterization, and molecular application of a virulence-associated autotransporter from a pathogenic Pseudomonas fluorescens strain [J]. Applied and Environmental Microbiology, 2009, 75(13): 4333—4340
[11]	Kristin S, Emina C, Elke H, et al. Molecular and proteome analyses highlight the importance of the Cpx envelope stress system for acid stress and cell wall stability in Escherichia coli [J]. Microbiology Open, 2016, 5(4): 582—596
[12]	李爱华. 我国鱼类病原菌耐药性、耐药质粒及几种药物抗菌作用的研究. 中国科学院水生生物研究所博士学位论文. 武汉. 1998 Li A H. The study of the drug resistance, resistance plasmid of fish pathogenic bacteria, and the antibacterial action of several drug in China [D]. Ph D thesis, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan. 1998
[13]	Acosta N, Pukatzki S, Raivio T L. The Vibrio cholerae Cpx envelope stress response senses and mediates adaptation to low iron [J]. Journal of Bacteriology, 2015, 197: 262—276
[14]	Raivio T L, Leblanc S K, Price N L, et al. The Escherichia coli Cpx envelope stress response regulates genes of diverse function that impact antibiotic resistance and membrane integrity [J]. Journal of Bacteriology, 2013, 195: 2755—2767
[15]	Livak K J, Schmittgen T D. Analysis of relative gene expression data using real-time quantitative PCR and the 2^(–ΔΔCt) method [J]. Methods, 2001, 25(4): 402—408
[16]	McEwen J, Silverman P. Chromosomal mutations of Escherichia coli that alter expression of conjugative plasmid functions [J]. Proceedings of the National Academy of Sciences of the United States of America, 1980, 77: 513—517
[17]	Cosma C L, Danese P N, Carlson J H, et al. Mutational activation of the Cpx signal transduction pathway of Escherichia coli suppresses the toxicity conferred by certain envelope-associated stresses [J]. Molecular Microbiology, 1995, 18: 491—505
[18]	De W P, Lin E C. Cpx Two-Component Signal Transduction in Escherichia coli: Excessive CpxR-P levels underlie CpxA* phenotypes [J]. Journal of Bacteriology, 2000, 182(5): 1423—1426
[19]	De W P, Kwon O, Lin E C. The CpxRA signal transduction system of Escherichia coli: Growth-related autoactivation and control of unanticipated target operons [J]. Journal of Bacteriology, 1999, 181(21): 6772—6778
[20]	Gal-Mor O, Segal G. Identification of cpxR as a positive regulator of icm and dot virulence genes of Legionella pneumophila [J]. Journal of Bacteriology, 2003, 185(16): 4908—4919
[21]	Jubelin G, Vianney A, Beloin C, et al. Cpx R/Omp R interplay regulates curli gene expression in response to osmolarity in Escherichia coli [J]. Journal of Bacteriology, 2005, 187(6): 2038—2049
[22]	Gerken H, Misra R. MzrA-EnvZ interactions in the periplasm influence the EnvZ/OmpR two-component regulon [J]. Journal of Bacteriology, 2010, 192(23): 6271—6278
[23]	Blanvillain S, Meyer D, Boulanger A, et al. Plant carbohydrate scavenging through tonB-dependent receptors: a feature shared by phytopathogenic and aquatic bacteria [J]. PLoS One, 2007, 2: e224
[24]	Schauer K, Gouget B, Carrière M, et al. Novel nickel transport mechanism across the bacterial outer membrane energized by the TonB/ExbB/ExbD machinery [J]. Molecular Microbiology, 2007, 63: 1054—1068
[25]	Humphreys S, Rowley G, Stevenson A, et al. Role of the two-component regulator Cpx AR in the virulence of Salmonella entetica serotype typhimurium [J]. Infection and Immunity, 2004, 72(8): 4654—4661
[26]	Debnath I, Norton J P, Barber A E, et al. The Cpx stress response system potentiates the fitness and virulence of uropathogenic Escherichia coli [J]. Infection and Immunity, 2013, 81(5): 1450—1459

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CONSTRUCTION AND BIOLOGICAL CHARACTERISTICS ANALYSIS OF CPXRA MUTANT IN AEROMONAS HYDROPHILA

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