鼠伤寒沙门菌sapC基因缺失株的构建及生物学特性分析

doi:10.11843/j.issn.0366-6964.2022.07.024

Abstract

Abstract: This study aimed to study the function of sapC in ABC transporters of SalmonellaTyphimurium in its pathogenic mechanism. The sapC gene deletion mutant SM△sapC of S. Typhimurium was constructed by λ-Red mediated mutagenesis procedures, and the growth characteristics, acid stress test, polymyxin B sensitivity test, biofilm detection, intracellular survival and toxicity test in mice were performed. The results showed that the growth rate of the deletion strain SMΔsapC was not significantly different from that of the parent strain and the supplemented strain. Under acid stress condition, the survival rate of SMΔsapCsapC was significantly lower than that of the parent strain; the deletion of sapC gene enhanced the drug resistance of polymyxin B and decreased the ability of biofilm formation of S. Typhimurium. The proliferation ability of SMΔsapC strain in mouse macrophages and the virulence in mice were significantly lower than that of the parent strain. This study showed sapC gene decreased the acid resistance and biofilm formation ability of S. Typhimurium, thus reducing the virulence of Salmonella in vivo and in vitro. This study laid a foundation for further explaining the pathogenic mechanism of S.Typhimurium.

Key words: Salmonella Typhimurium, homologous recombination, sapC, biological characteristics

CLC Number:

S852.61

CAO Li, CHENG Runan, WU Zhouhui, WANG Jiawei, WANG Yu, ZHANG Yonghong, WU Qingmin, WANG Zhen. Construction and Biological Characteristics of sapC Gene Deletion Strain of Salmonella Typhimurium[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(7): 2282-2289.

References 30

[1]	FUCHE F J, SOW O, SIMON R, et al. Salmonella Serogroup C:current status of vaccines and why they are needed[J]. Clin Vaccine Immunol, 2016, 23(9):737-745.
[2]	ANSARI N, YAZDIAN-ROBATI R, SHAHDORDIZADEH M, et al. Aptasensors for quantitative detection of Salmonella Typhimurium[J]. Anal Biochem, 2017, 533:18-25.
[3]	GARAI P, GNANADHAS D P, CHAKRAVORTTY D. Salmonella enterica serovars Typhimurium and Typhi as model organisms:revealing paradigm of host-pathogen interactions[J]. Virulence, 2012, 3(4):377-388.
[4]	SUN H H, WAN Y P, DU P C, et al. The epidemiology of monophasic Salmonella Typhimurium[J]. Foodborne Pathog Dis, 2020, 17(2):87-97.
[5]	QI X L, LI P, XU X G, et al. Epidemiological and molecular investigations on Salmonella responsible for gastrointestinal infections in the Southwest of Shanghai from 1998 to 2017[J]. Front Microbiol, 2019, 10:2025.
[6]	HANCOCK R E W, SCOTT M G. The role of antimicrobial peptides in animal defenses[J]. Proc Natl Acad Sci U S A, 2000, 97(16):8856-8861.
[7]	DE CESARE C B, CRISTY S A, GARSIN D A, et al. Antimicrobial peptides:a new frontier in antifungal therapy[J]. mBio, 2020, 11(6):e02123-20.
[8]	GALLO R L, MURAKAMI M, OHTAKE T, et al. Biology and clinical relevance of naturally occurring antimicrobial peptides[J]. J Allergy ClinImmunol, 2002, 110(6):823-831.
[9]	BADER M W, NAVARRE W W, SHIAU W, et al. Regulation of Salmonella typhimurium virulence gene expression by cationic antimicrobial peptides[J]. Mol Microbiol, 2003, 50(1):219-230.
[10]	BRODSKY I E, ERNST R K, MILLER S I, et al. mig-14 is a Salmonella gene that plays a role in bacterial resistance to antimicrobial peptides[J]. J Bacteriol, 2002, 184(12):3203-3213.
[11]	RODAS P I, CONTRERAS I, MORA G C. Salmonella enterica serovar Typhi has a 4. 1 kb genetic island inserted within the sapABCDF operon that causes loss of resistance to the antimicrobial peptide protamine[J]. J Antimicrob Chemother, 2010, 65(8):1624-1630.
[12]	PARRA-LOPEZ C, NAER M T, GROISMAN E A. Molecular genetic analysis of a locus required for resistance to antimicrobial peptides in Salmonella typhimurium[J]. EMBO J, 1993, 12(11):4053-4062.
[13]	ROEPE P D, WEI L Y, HOFFMAN M M, et al. Altered drug translocation mediated by the MDR protein:direct, indirect, or both?[J]. J Bioenerg Biomembr, 1996, 28(6):541-555.
[14]	KRAUSS S, ZIPPERER A, WIRTZ S, et al. Secretion of and self-resistance to the novel fibupeptide antimicrobial Lugdunin by distinct ABC transporters in Staphylococcus lugdunensis[J]. Antimicrob Agents Chemother, 2020, 65(1):e01734-20.
[15]	YAMAGISHI A, NAKANO S, YAMASAKI S, et al. An efflux inhibitor of the MacAB pump in Salmonella enterica serovar typhimurium[J]. Microbiol Immunol, 2020, 64(3):182-188.
[16]	曹莉,孙路,孔梓安,等.肠炎沙门氏菌yjeA基因缺失株的构建及其生物学功能测定[J].北京农学院学报, 2020, 35(2):16-20.CAO L, SUN L, KONG Z A, et al. Construction and characterization of yjeA gene deletion strain of Salmonella enteritidis[J]. Journal of Beijing University of Agriculture, 2020, 35(2):16-20.
[17]	ARANDA C M A, SWANSON J A, LOOMIS W P, et al. Salmonella Typhimurium activates virulence gene transcription within acidified macrophage phagosomes[J]. Proc Natl Acad Sci U S A, 1992, 89(21):10079-10083.
[18]	GUTIÉRREZ S, FISCHER J, GANESAN R, et al. Salmonella Typhimurium impairs glycolysis-mediated acidification of phagosomes to evade macrophage defense[J]. PLoS Pathog, 2021, 17(9):e1009943.
[19]	YUKI K E, EVA M M, RICHER E, et al. Suppression of hepcidin expression and iron overload mediate Salmonella susceptibility in Ankyrin 1 ENU-induced mutant[J]. PLoS One, 2013, 8(2):e55331.
[20]	PAUDYAL N, PAN H, WU B B, et al. Persistent asymptomatic human infections by Salmonella enterica serovar Newport in China[J]. mSphere, 2020, 5(3):e00163-20.
[21]	COBBOLD R N, RICE D H, DAVIS M A, et al. Long-term persistence of multi-drug-resistant Salmonella enterica serovar Newport in two dairy herds[J]. J Am Vet Med Assoc, 2006, 228(4):585-591.
[22]	ZHAO X X, DAI Q L, ZHU D K, et al. Recombinant attenuated Salmonella Typhimurium with heterologous expression of the Salmonella Choleraesuis O-polysaccharide:high immunogenicity and protection[J]. Sci Rep, 2017, 7(1):7127.
[23]	KUMAR A, ALAM A, RANI M, et al. Biofilms:survival and defense strategy for pathogens[J]. Int J Med Microbiol, 2017, 307(8):481-489.
[24]	DULA S, AJAYEOBA T A, IJABADENIYI O A. Bacterial biofilm formation on stainless steel in the food processing environment and its health implications[J]. Folia Microbiol (Praha), 2021, 66(3):293-302.
[25]	CHEBOTAR'I V, KONCHAKOVA E D, EVTEEVA N I. Neutrophil dependent breakdown of biofilms formed by Staphylococcus aureus[J]. Zh Mikrobiol Epidemiol Immunobiol, 2012(1):10-15.
[26]	MANGALAPPALLI-ILLATHU A K, KORBER D R. Adaptive resistance and differential protein expression of Salmonella enterica serovar Enteritidis biofilms exposed to benzalkonium chloride[J]. Antimicrob Agents Chemother, 2006, 50(11):3588-3596.
[27]	CHIN K C J, TAYLOR T D, HEBRARD M, et al. Transcriptomic study of Salmonella enterica subspecies enterica serovar Typhi biofilm[J]. BMC Genomics, 2017, 18:836.
[28]	HU S F, YU Y G, LV Z Q, et al. Proteomics study unveils ROS balance in acid-adapted Salmonella Enteritidis[J]. Food Microbiol, 2020, 92:103585.
[29]	JIAO S S, ZHANG H J, LIAO M J, et al. Investigation of the potential direct and cross protection effects of sublethal injured Salmonella Typhimurium induced by radio frequency heating stress[J]. Food Res Int, 2021, 150:110789.
[30]	ARVIZU-MEDRANO S M, ESCARTÍN E F. Effect of acid shock with hydrochloric, citric, and lactic acids on the survival and growth of Salmonella typhi and Salmonella typhimurium in acidified media[J]. J Food Prot, 2005, 68(10):2047-2053.

Construction and Biological Characteristics of sapC Gene Deletion Strain of Salmonella Typhimurium

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References 30

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	LI Zhaoyan, GAO Jiang, GUO Shihui, ZHAO Ruqian, MA Wenqiang. Advances in Detection Methods and Control Solutions for Feline Allergens [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(6): 2272-2279.
[2]	LIU Qing, WU Shaopeng, SHI Bin, SHAO Hongxia, QIAN Kun, YE Jianqiang, QIN Aijian. Expression of Chicken CTLA-4 Protein in Insect Cells and the Preparation of Its Monoclonal Antibody [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(6): 2596-2604.
[3]	QIN Lei, WU Huimin, XU Qiqi, CHEN Wanzhao, WANG Dong, LI Hongbo, XIA Panpan, LIU Zepeng, XIA Lining. Effect of Exogenous Drug-Resistant Salmonella Typhimurium on Intestinal Flora in Healthy Mice [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(5): 2158-2169.
[4]	JIANG Zenghai, TENG Lin, HE Anwen, LIU Yanyan, YUE Min, HE Qigai. Genomic Analysis of Salmonella Typhimurium Isolates and Salmonella Serotype 4, [5], 12: i:- Isolates from Pig-borne Food Chain [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(3): 1199-1209.
[5]	LI Xiaohan, YANG Fuchun, LIU Rui, GAO Li, CUI Hongyu, ZHANG Yanping, LIU Changjun, QI Xiaole, WANG Xiaomei, GAO Yulong, LI Kai. Construction of Recombinant Herpesvirus of Turkey Expressing Infectious Bursal Disease Virus VP2 Gene and Analysis of Its Biological Characteristics [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(2): 656-662.
[6]	LI Lili, CHEN Kaifeng, CHEN Bing, ZHOU Zhouping, WANG Nanwei, QU Xiaoyun, XU Chenggang, LIAO Ming, ZHANG Jianmin. Regulatory Role of STM1827 in the Biofilm Formation and Environmental Stress of Salmonella Typhimurium [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(12): 5207-5217.
[7]	HAN Shengyi, LI Lingxia, LI Shuping, HU Guoyuan, LI Shengqing. Biological Characteristics and Genome Analysis of Salmonella Phage SP3 [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(12): 5228-5239.
[8]	CEN Xin, YANG Tingting, ZHAO Zunfu, WEN Yongping, ZHANG Huanrong. Isolation, Identification, Biological Characteristics and Genome Analysis of a Virulent Salmonella Phage [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(8): 2677-2688.
[9]	LIU Chang, LIU Jiao, CHEN Jin, WANG Mianzhi, XIONG Wenguang, ZENG Zhenling. Isolation and Biological Characteristics of a Bacteriophage Capable of Lysing bla_NDM-5 and mcr-1 Positive Escherichia coli [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(3): 857-866.
[10]	ZHANG Lin, LU Fang, FU Hengfeng, JIANG Xidi, WEI Qiling, QI Caili, GAO Haixia, LI Lin. Regulation Mechanism of BaeSR on Fluoroquinolones Resistance in Salmonella Typhimurium [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(3): 894-903.
[11]	SUN Peihao, ZHAO Xinzhe, WU Hanxiao, LÜ Ce, YANG Liguo, LIANG Aixin. Primary Culture and Biological Characteristics Analysis of Bovine Mammary Epithelial Cells [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(2): 447-458.
[12]	WANG Yunan, WU Yue, SONG Hanan, ZHANG Tao, GUAN Weijun. Primary Culture of Simmental Bovine Pancreatic Mesenchymal Stem Cells and Their Multi-directional Differentiation Potential [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(11): 3786-3796.
[13]	LINGHU Yuanfeng, PAN Yong, YANG Yang, DUAN Shiyu, ZHANG Jiali, ZHANG Baotai, YANG Qi. Preliminary Exploration of Binding Sites of Salmonella Typhimurium Chaperone Protein Hfq on Small RNA GcvB [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(11): 4110-4115.
[14]	WU Liting, LIU Banhong, TIAN Yuan, WANG Juan, BAO Hongduo, ZHOU Yan, PANG Maoda, WANG Ran, ZHANG Hui. Identification and Environmental Inhibition Effect of Clostridium perfringens Phage [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(10): 3550-3560.
[15]	SHAO Ying, FU Dandan, WU Xiaoyan, GU Yi, SONG Xiangjun, TU Jian, QI Kezong. Effect of the ETT2 Structural Gene epaPQR on the Biological Properties and Pathogenicity of Avian Pathogenic Escherichia coli [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(10): 3631-3641.