Pelludat Cosima, Hogardt Michael, Heesemann Jürgen
Max von Pettenkofer-Institut für Hygiene und Medizinische Mikrobiologie, 80336 Munich, Germany.
Infect Immun. 2002 Apr;70(4):1832-41. doi: 10.1128/IAI.70.4.1832-1841.2002.
The high-pathogenicity island (HPI) of yersiniae encodes an iron uptake system represented by its siderophore yersiniabactin (Ybt). The HPI is present in yersiniae with high levels of pathogenicity--i.e., Yersinia pestis, Y. pseudotuberculosis, and Y. enterocolitica biogroup (BG) 1B--but absent in Y. enterocolitica strains with low (BG 2 to 5) and no (BG 1A) levels of pathogenicity and has been shown to be an important virulence factor. Comparison of the HPI in Y. enterocolitica (Yen-HPI) and that in Y. pestis and Y. pseudotuberculosis revealed that, in contrast to genes of the variable region, genes of the core region (genes irp9 to fyuA) are highly homologous. In the present work the Yen-HPI core genes were rescued from the chromosome of Y. enterocolitica WA-C (BG 1B, serotype O:8) using the FRT-FLP recombinase system. Transfer of the resulting plasmid pCP1 into the siderophore-deficient strain Y. enterocolitica NF-O (BG 1A) led to no halo on siderophore indicator chrome azurol S (CAS) agar. Transfer of pCP1 into the Y. enterocolitica strain MRS40 (serotype O:9, BG 2; phenotype, CAS negative) led to a CAS halo larger than that of parental strain WA-C, indicating high Ybt production. pCP1 was highly unstable in iron-deficient medium, and no enhanced mouse virulence conferred by MRS40 carrying pCP1 could be detected. To overcome the problem of instability, pCP1 was integrated into the chromosome of MRS40, leading to the formation of a CAS halo comparable to that seen with WA-C and correspondingly to increased mouse virulence. Thus, the core genes of Yen-HPI are sufficient to confer a positive CAS phenotype and mouse virulence to Y. enterocolitica MRS40, BG 2, but are insufficient to confer this phenotype to Y. enterocolitica NF-O, BG 1A.
耶尔森菌的高致病性岛(HPI)编码一种以其铁载体耶尔森菌素(Ybt)为代表的铁摄取系统。HPI存在于致病性高的耶尔森菌中,即鼠疫耶尔森菌、假结核耶尔森菌和小肠结肠炎耶尔森菌生物群(BG)1B,但在致病性低(BG 2至5)和无致病性(BG 1A)的小肠结肠炎耶尔森菌菌株中不存在,并且已被证明是一种重要的毒力因子。对小肠结肠炎耶尔森菌(Yen-HPI)与鼠疫耶尔森菌和假结核耶尔森菌中的HPI进行比较发现,与可变区基因不同,核心区基因(irp9至fyuA基因)具有高度同源性。在本研究中,使用FRT-FLP重组酶系统从小肠结肠炎耶尔森菌WA-C(BG 1B,血清型O:8)的染色体中拯救出Yen-HPI核心基因。将所得质粒pCP1转入铁载体缺陷型菌株小肠结肠炎耶尔森菌NF-O(BG 1A)后,在铁载体指示铬天青S(CAS)琼脂上未产生晕圈。将pCP1转入小肠结肠炎耶尔森菌菌株MRS40(血清型O:9,BG 2;表型,CAS阴性)后,产生的CAS晕圈比亲本菌株WA-C的大,表明Ybt产量高。pCP1在缺铁培养基中高度不稳定,并且未检测到携带pCP1的MRS40增强的小鼠毒力。为克服不稳定性问题,将pCP1整合到MRS40的染色体中,导致形成与WA-C相当的CAS晕圈,并相应增加了小鼠毒力。因此,Yen-HPI的核心基因足以赋予小肠结肠炎耶尔森菌MRS40(BG 2)阳性CAS表型和小鼠毒力,但不足以赋予小肠结肠炎耶尔森菌NF-O(BG 1A)这种表型。