蜡样芽胞杆菌(Bacillus cereus)是一种常见的食源性致病菌。从食物中毒病人的食物残渣中分离到1株产呕吐毒素的蜡样芽胞杆菌BC307,其对青霉素类抗生素具有较强的抗性。抗SigP因子RsiP是蜡样芽胞杆菌族(Bacillus cereus sensu lato)细菌青霉素耐药的重要调控因子。该实验利用原核系统表达RsiP蛋白并利用亲和层析法进行纯化,免疫小鼠后获得高效价多克隆抗体。结果表明,RsiP蛋白在炭疽芽胞杆菌、蜡样芽胞杆菌、苏云金芽胞杆菌中高度同源,在Western Blotting实验中RsiP抗血清与这3种菌的全菌蛋白均能发生特异性免疫反应。研究结果为探究RsiP对蜡样芽胞杆菌群细菌的青霉素耐药性调控打下了良好的基础。
Bacillus cereus is a common food-borne pathogen and anti-SigP factor RsiP is an important regulator of penicillin resistance. B. cereus BC307 strain from the food residues of food poisoning patient was isolated, which has strong resistance to penicillin. The RsiP protein was expressed and purified by affinity chromatography. High-titer polyclonal antibody was obtained after immunizing mice. The results showed that the RsiP protein was highly conserved and identical in B. anthracis, B. cereus, and B. thuringiensis. Western blotting showed that the RsiP antiserum had specific immunological reactions with the whole bacterial protein of these three bacteria. The studies lay a good foundation for exploring the regulation of penicillin resistance of B. cereus by RsiP.
[1] DIETRICH R,JESSBERGER N,EHLING-SCHULZ M,et al.The food poisoning toxins of Bacillus cereus[J].Toxins,2021,13(2):98.
[2] NARANJO M,DENAYER S,BOTTELDOORN N,et al.Sudden death of a young adult associated with Bacillus cereus food poisoning[J].Journal of Clinical Microbiology,2011,49(12):4 379-4 381.
[3] RAJKOVIC A,UYTTENDAELE M,VERMEULEN A,et al.Heat resistance of Bacillus cereus emetic toxin,cereulide[J].Letters in Applied Microbiology,2008,46(5):536-541.
[4] MASSOVA I,MOBASHERY S.Structural and mechanistic aspects of evolution of β-lactamases and penicillin-binding proteins[J].Current Pharmaceutical Design,1999,5(11):929-937.
[5] HO T D,ELLERMEIER C D.Extra cytoplasmic function σ factor activation[J].Current Opinion in Microbiology,2012,15(2):182-188.
[6] ROSS C L,THOMASON K S,KOEHLER T M.An Extracytoplasmic function σ factor controls β-lactamase gene expression in Bacillus anthracis and other Bacillus cereus group species[J].Journal of Bacteriology,2009,191(21):6 683-6 693.
[7] HO T D,NAUTA K M,MÜH U,et al.Activation of the extracytoplasmic function σ factor σ(P) by β-lactams in Bacillus thuringiensis requires the site-2 protease RasP[J].mSphere,2019,4(4).DOI:10.1101/707133.
[8] HA J,PARK Y J,KIM Y J,et al.Late prosthetic joint infection and bacteremia by Bacillus cereus confirmed by 16S rRNA sequencing and hip joint tissue pathology[J].Annals of Clinical Microbiology,2016,19(2):54-57.
[9] WANG L T,LEE F L,TAI C J,et al.Comparison of gyrB gene sequences,16S rRNA gene sequences and DNA-DNA hybridization in the Bacillus subtilis group[J].International Journal of Systematic and Evolutionary Microbiology,2007,57:1 846-1 850.
[10] OKINAKA R T,CLOUD K,HAMPTON O,et al.Sequence and organization of pXO1,the large Bacillus anthracis plasmid harboring the Anthrax toxin genes[J].Journal of Bacteriology,1999,181(20):6 509-6 515.
[11] KOEHLER T M.Bacillus anthracis genetics and virulence gene regulation[J].Current Topics in Microbiology and Immunology,2002,271:143-164.
[12] 赵雨, 郭建华,张春枝.蜡状芽孢杆菌ZY12产磷脂酶D的影响因素[J].食品与发酵工业,2021,47(9):57-62.
ZHAO Y,GUO J H,ZHANG C Z.Factors influencing phospholipase D production by Bacillus cereus ZY12[J].Food and Fermentation Industries,2021,47(9):57-62.
[13] SENESI S,GHELARDI E.Production,secretion and biological activity of Bacillus cereus enterotoxins[J].Toxins (Basel),2010,2(7):1 690-1 703.
[14] LIU X K,QI X P,ZHU L,et al.Genome sequence of Bacillus anthracis attenuated vaccine strain A16R used for human in China[J].Journal of Biotechnology,2015,210:15-16.
[15] KUMAR S,STECHER G,LI M,et al.MEGA X:Molecular evolutionary genetics analysis across computing platforms[J].Molecular Biology and Evolution,2018,35(6):1 547-1 549.
[16] EHLING-SCHULZ M,FRICKER M,GRALLERT H,et al.Cereulide synthetase gene cluster from emetic Bacillus cereus:Structure and location on a mega virulence plasmid related to Bacillus anthracis toxin plasmid pXO1[J].BMC Microbiology,2006,6:20.
[17] BAZINET A L.Pan-genome and phylogeny of Bacillus cereus sensu lato[J].BMC Evolutionary Biology,2017,17(1):176.
[18] GARGIS A S,MCLAUGHLIN H P,CONLEY A B,et al.Analysis of whole-genome sequences for the prediction of penicillin resistance and β-lactamase activity in Bacillus anthracis[J].mSystems,2018,3(6).DOI:10.1128/msystems.00 154-18.
[19] PANAVAS T,SANDERS C,BUTT T R.SUMO fusion technology for enhanced protein production in prokaryotic and eukaryotic expression systems[J].Methods in Molecular Biology,2009,497:303-317.
[20] TAMURA K,NEI M,KUMAR S.Prospects for inferring very large phylogenies by using the neighbor-joining method[J].PNAS,2004,101(30):11 030-11 035.
[21] LÜCKING G,FRENZEL E,RüTSCHLE A,et al.Ces locus embedded proteins control the non-ribosomal synthesis of the cereulide toxin in emetic Bacillus cereus on multiple levels[J].Frontiers in Microbiology,2015,6:1101.
[22] TAKENO A,OKAMOTO A,TORI K,et al.Complete genome sequence of Bacillus cereus NC7401,which produces high levels of the emetic toxin cereulide[J].Journal of Bacteriology,2012,194(17):4 767-4 768.
[23] 韩蓓, 吕佳,杜仁佳.蜡状芽孢杆菌相关食物中毒研究进展[J].国外医学(医学地理分册),2017,38(3):211-214.
HAN B,LYU J,DU R J.Review on the food poisoning caused by Bacillus cereus[J].Foreign Medical Sciences(Section of Medgeography),2017,38(3):211-214.
