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Genetic Analysis of Bacteriocin 43 of Vancomycin-Resistant Enterococcus faecium

机译:耐万古霉素粪肠球菌细菌素43的遗传分析

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A total of 636 vancomycin-resistant Enterococcus faecium (VRE) isolates obtained between 1994 and 1999 from the Medical School Hospital of the University of Michigan were tested for bacteriocin production. Of the 277 (44%) bacteriocinogenic strains, 21 were active against E. faecalis, E. faecium, E. hirae, E. durans, and Listeria monocytogenes. Of those 21 strains, a representative bacteriocin of strain VRE82, designated bacteriocin 43, was found to be encoded on mobilizable plasmid pDT1 (6.2 kbp). Nine open reading frames (ORFs), ORF1 to ORF9, were presented on pDT1 and were oriented in the same direction. The bacteriocin 43 locus (bac43) consists of the bacteriocin gene bacA (ORF1) and the immunity gene bacB (ORF2). The deduced bacA product is 74 amino acids in length with a putative signal peptide of 30 amino acids at the N terminus. The bacB gene encodes a deduced 95-amino-acid protein without a signal sequence. The predicted mature BacA protein (44 amino acids) showed sequence homology with the membrane-active class IIa bacteriocins of lactic acid bacteria and showed 86% homology with bacteriocin 31 from E. faecalis YI717 and 98% homology with bacteriocin RC714. Southern analysis with a bac43 probe of each plasmid DNA from the 21 strains showed hybridization to a specific fragment corresponding to the 6.2-kbp EcoRI fragment, suggesting that the strains harbored the pDT1-like plasmid (6.2 kb) which encoded the bacteriocin 43-type bacteriocin. The bac43 determinant was not identified among non-VRE clinical isolates.
机译:测试了1994年至1999年间从密歇根大学医学院附属医院获得的636种耐万古霉素粪便肠球菌(VRE)分离株的细菌素产生。在277(44%)的细菌致癌菌株中,有21种对粪肠球菌,粪肠球菌,平肠埃希氏菌,杜兰氏大肠杆菌和单核细胞增生李斯特菌具有活性。在这21个菌株中,发现菌株VRE82的代表性细菌素,称为细菌素43,被编码在可动员质粒pDT1(6.2kbp)上。在pDT1上展示了9个开放阅读框(ORF),即ORF1至ORF9,它们的方向相同。细菌素43位点(bac43)由细菌素基因bacA(ORF1)和免疫基因bacB(ORF2)组成。推导的bacA产物长度为74个氨基酸,在N端带有30个氨基酸的推定信号肽。 bacB基因编码一个没有信号序列的推断的95个氨基酸的蛋白质。预测的成熟BacA蛋白(44个氨基酸)与乳酸菌的膜活性IIa类细菌素具有序列同源性,与粪肠球菌YI717的细菌素31具有86%的同源性,与细菌素RC714具有98%的同源性。用bac43探针对来自21个菌株的每种质粒DNA进行的Southern分析表明,该片段与对应于6.2-kbp EcoRI片段的特定片段杂交,表明该菌株带有pDT1样质粒(6.2 kb),该质粒编码细菌素43型细菌素。在非VRE临床分离株中未鉴定到bac43决定簇。

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