CHARACTERIZATION OF TN1546, A TN3-RELATED TRANSPOSON CONFERRING GLYCOPEPTIDE RESISTANCE BY SYNTHESIS OF DEPSIPEPTIDE PEPTIDOGLYCAN PRECURSORS IN ENTEROCOCCUS-FAECIUM BM4147
CHARACTERIZATION OF TN1546, A TN3-RELATED TRANSPOSON CONFERRING GLYCOPEPTIDE RESISTANCE BY SYNTHESIS OF DEPSIPEPTIDE PEPTIDOGLYCAN PRECURSORS IN ENTEROCOCCUS-FAECIUM BM4147
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DOI：
10.1128/jb.175.1.117-127.1993
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发表时间：
1993-01-01
影响因子：
3.2
通讯作者：
COURVALIN, P
中科院分区：
文献类型：
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作者：
ARTHUR, M;MOLINAS, C;COURVALIN, P
屎肠球菌BM 4147中pIP 816携带的万古霉素抗性货车基因簇的侧翼区的序列测定显示与Tn 3家族的转座子相似。不完全反向重复序列（38 bp中的36个）描绘了一个10，851 bp的元件，命名为Tn 1546。位于vanR基因上游的4-kb区域含有两个以相反方向转录的开放阅读框（OPF）。推导的ORFI的氨基酸序列（988个残基）显示，分别为56%和42%的同源性转座酶的Tn 4430从苏云金芽孢杆菌和Tn 917从粪肠球菌。ORF 2的产物（191个残基）与Tn 917的解离酶（33%氨基酸同一性）和产气荚膜梭菌质粒pIP 404的Res蛋白（48%）相关。Tn 1546在大肠杆菌中连续地从质粒pUC 18转座到pOX 38，并从pOX 38转座到pBR 329的各个位点。转座是复制性的，导致共整合体的形成，并在靶位点产生5-bp的重复。Southern杂交和DNA扩增揭示了Tn 1546相关元件的存在下，肠球菌高度耐糖肽。对这些元件周围序列的分析表明，转座在货车基因簇在人临床分离的大肠杆菌复制子中的传播中起作用。屎室
Sequence determination of the flanking regions of the vancomycin resistance van gene cluster carried by pIP816 in Enterococcus faecium BM4147 revealed similarity to transposons of the Tn3 family. Imperfect inverted repeats (36 of 38 bp) delineated a 10,851-bp element designated Tn1546. The 4-kb region located upstream from the vanR gene contained two open reading frames (OPF) transcribed in opposite directions. The deduced amino acid sequence of ORFI (988 residues) displayed, respectively, 56 and 42% identity to those of the transposases of Tn4430 from Bacillus thuringiensis and of Tn917 from Enterococcus faecalis. The product of ORF2 (191 residues) was related to the resolvase of Tn917 (33% amino acid identity) and to the Res protein (48%) of plasmid pIP404 from Clostridium perfringens. Tn1546 transposed consecutively in Escherichia coli from plasmid pUC18 into pOX38 and from pOX38 into various sites of pBR329. Transposition was replicative, led to the formation of cointegrates, and produced a 5-bp duplication at the target site. Southern hybridization and DNA amplification revealed the presence of Tn1546-related elements in enterococci highly resistant to glycopeptides. Analysis of sequences surrounding these elements indicated that transposition plays a role in dissemination of the van gene cluster among replicons of human clinical isolates of E. faecium.