Wang Yanping, Wang Gui-Rong, Shelby Aikiesha, Shoemaker Nadja B, Salyers Abigail A
Department of Microbiology, University of Illinois, Urbana, Illinois 61801, USA.
Appl Environ Microbiol. 2003 Aug;69(8):4595-603. doi: 10.1128/AEM.69.8.4595-4603.2003.
Results of a recent study of antibiotic resistance genes in human colonic Bacteroides strains suggested that gene transfer events between members of this genus are fairly common. The identification of Bacteroides isolates that carried an erythromycin resistance gene, ermG, whose DNA sequence was 99% identical to that of an ermG gene found previously only in gram-positive bacteria raised the further possibility that conjugal elements were moving into Bacteroides species from other genera. Six of seven ermG-containing Bacteroides strains tested were able to transfer ermG by conjugation. One of these strains was chosen for further investigation. Results of pulsed-field gel electrophoresis experiments showed that the conjugal element carrying ermG in this strain is an integrated element about 75 kb in size. Thus, the element appears to be a conjugative transposon (CTn) and was designated CTnGERM1. CTnGERM1 proved to be unrelated to the predominant type of CTn found in Bacteroides isolates-CTns of the CTnERL/CTnDOT family-which sometimes carry another type of erm gene, ermF. A 19-kbp segment of DNA from CTnGERM1 was cloned and sequenced. A 10-kbp portion of this segment hybridized not only to DNA from all the ermG-containing strains but also to DNA from strains that did not carry ermG. Thus, CTnGERM1 seems to be part of a family of CTns, some of which have acquired ermG. The percentage of G+C content of the ermG region was significantly lower than that of the chromosome of Bacteroides species-an indication that CTnGERM1 may have entered Bacteroides strains from some other bacterial genus. A survey of strains isolated before 1970 and after 1990 suggests that the CTnGERM1 type of CTn entered Bacteroides species relatively recently. One of the genes located upstream of ermG encoded a protein that had 85% amino acid sequence identity with a macrolide efflux pump, MefA, from Streptococcus pyogenes. Our having found >90% sequence identity of two upstream genes, including mefA, and the remnants of two transposon-carried genes downstream of ermG with genes found previously only in gram-positive bacteria raises the possibility that gram-positive bacteria could have been the origin of CTnGERM1.
最近一项关于人类结肠拟杆菌菌株中抗生素抗性基因的研究结果表明,该属成员之间的基因转移事件相当普遍。对携带红霉素抗性基因ermG的拟杆菌分离株的鉴定显示,其DNA序列与之前仅在革兰氏阳性菌中发现的ermG基因的序列有99%的同一性,这进一步增加了接合元件从其他属转移到拟杆菌属物种中的可能性。所测试的七个含有ermG的拟杆菌菌株中有六个能够通过接合作用转移ermG。选择其中一个菌株进行进一步研究。脉冲场凝胶电泳实验结果表明,该菌株中携带ermG的接合元件是一个大小约为75 kb的整合元件。因此,该元件似乎是一个接合转座子(CTn),并被命名为CTnGERM1。事实证明,CTnGERM1与在拟杆菌分离株中发现的主要类型的CTn(CTnERL/CTnDOT家族的CTn)无关,后者有时携带另一种erm基因ermF。从CTnGERM1中克隆并测序了一段19 kbp的DNA片段。该片段的一个10 kbp部分不仅与所有含有ermG的菌株的DNA杂交,也与不携带ermG的菌株的DNA杂交。因此,CTnGERM1似乎是CTn家族的一部分,其中一些已经获得了ermG。ermG区域的G+C含量百分比显著低于拟杆菌属物种的染色体,这表明CTnGERM1可能是从其他细菌属进入拟杆菌菌株的。对1970年之前和1990年之后分离的菌株进行的一项调查表明,CTn的CTnGERM1类型是相对较新进入拟杆菌属物种的。位于ermG上游的一个基因编码一种蛋白质,该蛋白质与化脓性链球菌的大环内酯流出泵MefA具有85%的氨基酸序列同一性。我们发现包括mefA在内的两个上游基因以及ermG下游两个转座子携带基因的残余部分与之前仅在革兰氏阳性菌中发现的基因有>90%的序列同一性,这增加了革兰氏阳性菌可能是CTnGERM1起源的可能性。