Correia A, Pisabarro A, Castro J M, Martín J F
Dpt. of Ecology, Genetics and Microbiology, Faculty of Biology, University of León, Spain.
Gene. 1996 Apr 17;170(1):91-4. doi: 10.1016/0378-1119(95)00866-7.
A repetitive DNA element of the Gram+ Brevibacterium lactofermentum (Bl), cloned by a modification of the subtractive hybridization method, contained a 1.4-kb IS-like element, IS13869, which included an open reading frame (ORF) inside a perfect 26-bp terminal inverted repeat (TIR). An 8-bp direct repeat (DR) was found outside each TIR. The ORF encoded a deduced protein of 436 amino acids (49 380 Da) with extensive similarity to other known transposases of insertion elements of Mycobacterium smegmatis (IS1096). Pseudomonas sp. (tpnA) and Corynebacterium glutamicum (IS31831). Distinct patterns were observed in different strains of Bl by hybridization with a probe internal to IS13869: four copies of IS13869 occurred in the wild type (wt) and R31 strains, but only three of them were observed in a recA derivative of the wt. Analysis by pulsed-field gel electrophoresis suggested that at least one copy of IS13869 had changed its position inside the chromosome during the lineage of a Bl derivative.
通过对消减杂交方法进行改进克隆得到的革兰氏阳性乳酸发酵短杆菌(Bl)的一个重复DNA元件,包含一个1.4kb的类插入序列元件IS13869,其在一个完美的26bp末端反向重复序列(TIR)内包含一个开放阅读框(ORF)。在每个TIR外侧发现了一个8bp的正向重复序列(DR)。该ORF编码一个推导的436个氨基酸(49380Da)的蛋白质,与耻垢分枝杆菌(IS1096)、假单胞菌属(tpnA)和谷氨酸棒杆菌(IS31831)的插入元件的其他已知转座酶具有广泛的相似性。通过用IS13869内部的探针进行杂交,在不同的Bl菌株中观察到了不同的模式:野生型(wt)和R31菌株中有四个IS13869拷贝,但在wt的recA衍生物中只观察到其中三个。脉冲场凝胶电泳分析表明,在Bl衍生物的谱系中,至少有一个IS13869拷贝在染色体内改变了其位置。