Tauch A, Kassing F, Kalinowski J, Pühler A
Department of Genetics, University of Bielefeld, Germany.
Plasmid. 1995 Sep;34(2):119-31. doi: 10.1006/plas.1995.9995.
Analysis of the 50-kb R-plasmid pTP10 from the clinical isolate Corynebacterium xerosis M82B revealed that the erythromycin resistance gene, ermCX, is located on a 4524-bp composite transposable element, Tn5432. The ends of Tn5432 are identical, direct repeats of an insertion sequence, designated IS1249, encoding a putative transposase of the IS256 family. IS1249 consists of 1385 bp with 45/42 imperfect terminal inverted repeats. The nucleotide sequence of the 1754-bp Tn5432 central region is 99% identical to the previously sequenced erythromycin resistance region of the Corynebacterium diphtheriae plasmid pNG2. It encodes the erythromycin resistance gene, ermCX, and an ORF homologous to the amino-terminal end of the transposase of IS31831 from Corynebacterium glutamicum. Transposons with regions flanking the insertion sites were recovered from the C. glutamicum chromosome by a plasmid rescue technique. Insertion of Tn5432 created 8-bp target site duplications. A Tn5432-induced isoleucine/valine-auxotrophic mutant was found to carry the transposon in the 5' region of the ilvBNC cluster; in pTP10 the transposon is inserted in a region similar to replication and partitioning functions of the Enterococcus faecalis plasmid pAD1 and the Agrobacterium tumefaciens plasmid pTAR.
对临床分离株干燥棒状杆菌M82B的50 kb R质粒pTP10进行分析发现,红霉素抗性基因ermCX位于一个4524 bp的复合转座元件Tn5432上。Tn5432的末端是相同的,是一个插入序列的正向重复,命名为IS1249,编码IS256家族的一个假定转座酶。IS1249由1385 bp组成,具有45/42个不完全末端反向重复。Tn5432中心区域1754 bp的核苷酸序列与先前测序的白喉棒状杆菌质粒pNG2的红霉素抗性区域99%相同。它编码红霉素抗性基因ermCX,以及一个与谷氨酸棒状杆菌IS31831转座酶氨基末端同源的开放阅读框。通过质粒拯救技术从谷氨酸棒状杆菌染色体上回收了插入位点侧翼区域的转座子。Tn5432的插入产生了8 bp的靶位点重复。发现一个Tn5432诱导的异亮氨酸/缬氨酸营养缺陷型突变体在ilvBNC簇的5'区域携带转座子;在pTP10中,转座子插入到一个类似于粪肠球菌质粒pAD1和根癌土壤杆菌质粒pTAR的复制和分配功能的区域。