肠炎沙门菌iroB基因缺失株的构建及部分特性分析
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摘要
为研究糖基转移酶IroB对沙门菌的重要作用,本研究利用λ-Red重组技术构建了肠炎沙门菌iroB基因缺失突变株c50336ΔiroB,通过应激试验、胞内存活试验和药物耐受试验检测缺失iroB基因后肠炎沙门菌生物特性的变化。结果显示,iroB基因缺失并未影响细菌的生长速度和生化特性,也不影响其对酸、碱和过氧化氢的抵抗能力,以及其在细胞内的存活能力,但使得细菌对热应激的敏感性增加,对蛋清抵抗能力下降,推测IroB参与沙门菌对铁的利用和对多溶菌酶的耐受;利用多粘菌素B进行最低抑菌浓度试验,结果显示其耐该药能力降低。本研究结果揭示IroB能够促进肠炎沙门菌的抗应激能力和耐药性,阐释了糖基转移酶IroB在沙门菌致病过程中的作用。
To investigate the roles of C-glycosyltransferase IroB of Salmonella enterica, the iroB gene-knockout strain of S.enterica was constructed by λ-Red recombination, and its contribution to pathogenic mechanism of S.enterica was analyzed by stress test, drug sensitive test and cell infection test. The data demonstrated that, the iroB-deficient mutant showed no difference in growth characteristics, biochemical characteristics, intracellular survival ability and acid, alkali and oxidative stress, compared with wild type strain. However, the mutant strain showed increased sensibility to heat stress and survival defection in albumen. The possible molecular mechanisms is that IroB contribute to the survival of S.enterica under iron-limited conditions and in lysozyme.The results revealed IroB contributed to the anti-stress ability and drug resistence and explored underlying the role of IroB on the pathogenidty of S.enterica.
To investigate the roles of C-glycosyltransferase IroB of Salmonella enterica, the iroB gene-knockout strain of S.enterica was constructed by λ-Red recombination, and its contribution to pathogenic mechanism of S.enterica was analyzed by stress test, drug sensitive test and cell infection test. The data demonstrated that, the iroB-deficient mutant showed no difference in growth characteristics, biochemical characteristics, intracellular survival ability and acid, alkali and oxidative stress, compared with wild type strain. However, the mutant strain showed increased sensibility to heat stress and survival defection in albumen. The possible molecular mechanisms is that IroB contribute to the survival of S.enterica under iron-limited conditions and in lysozyme.The results revealed IroB contributed to the anti-stress ability and drug resistence and explored underlying the role of IroB on the pathogenidty of S.enterica.
引文
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[2]Peng Meng-fei,Salaheen S,Buchanan R L,et al.Alterations of Salmonella typhimurium antibiotic resistance under environmental pressure[J].Appl Environ Microbiol,2018,84(19):1-35.
[3]Ibrahim H R,Sugimoto Y,Aoki T.Ovotransferrin antimicrobial peptide(OTAP-92)kills bacteria through a membrane damage mechanism[J].Biochim Biophys Acta,2000,1523(2-3):196-205.
[4]刘谢,柯碧霞,何冬梅,等.2007~2013年广东省肠炎沙门菌耐药情况分析[J].中华疾病控制杂志,2015,(03):269-272.
[5]Mathew E N,Muyyarikkandy M S,Kuttappan D,et al.Attachment of Salmonella enterica on mangoes and survival under conditions simulating commercial mango packing house and importer facility[J].Front Microbiol,2018,9(1519):1-15.
[6]Srikumar S,Kroger C,Hebrard M,et al.RNA-seq brings new insights to the intra-macrophage transcriptome of Salmonella typhimurium[J].PLo S Pathog,2015,11(11):e1005262.
[7]蒋惠源,曹鸣辉,曹学松,等.非编码小RNA(sraB)调控肠炎型沙门菌的抗蛋清抑菌作用[J].微生物学报,2010,50(11):1537-1544.
[8]Namakchian M,Kassler K,Sticht H,et al.Structure-based functional analysis of effector protein SifA in living cells reveals motifs important for Salmonella intracellular proliferation[J].Int JMed Microbiol,2017,308(1):84-96.
[9]姚丰华,董立伟,张钰,等.肠炎沙门菌hcp基因对生物被膜形成的影响分析[J].中国预防兽医学报,2014,36(07):534-537.
[10]刘松艳,周薇,司微,等.肠炎沙门菌延伸因子EF-Tu缺失株对环境压力的应答变化[J].中国预防兽医学报,2016,38(01):31-34.
[11]Lu Sang-wei,Killoran P B,Riley L W.Association of Salmonella enterica serovar enteritidis yafD with resistance to chicken egg albumen[J].Infect Immun,2003,71(12):6734-6741.
[12]Shah D H,Zhou Xiao-hui,Kim H Y,et al.Transposon mutagenesis of Salmonella enterica serovar Enteritidis identifies genes that contribute to invasiveness in human and chicken cells and survival in egg albumen[J].Infect Immun,2012,80(12):4203-4215.
[13]Guo Rong-xian,Li Zhuo-yang,Jiao Yang,et al.O-polysaccharide is important for Salmonella pullorum survival in egg albumen,and virulence and colonization in chicken embryos[J].Avian Pathol,2017,46(5):535-540.
[14]Foshag D,Campbell C,Pawelek P D.The C-glycosyltransferase Iro B from pathogenic Escherichia coli:identification of residues required for efficient catalysis[J].Biochim Biophys Acta,2014,1844(9):1619-1630.
[15]Kerrinnes T,Young B M,Leon C,et al.Phospholipase A1modulates the cell envelope phospholipid content of Brucella melitensis,contributing to polymyxin resistance and pathogenicity[J].Antimicrob Agents Chemother,2015,59(11):6717-6724.
[16]陈福,罗玉萍,龚熹,等.抗菌肽耐药性研究进展[J].微生物学通报,2008,(11):1786-1790.
[17]Wang Zhen,Bie Peng-fei,Cheng Jie,et al.The ABC transporter YejABEF is required for resistance to antimicrobial peptides and the virulence of Brucella melitensis[J].Sci Rep,2016,6:31876.
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