Byrne A M, Lessie T G
Department of Microbiology, University of Massachusetts, Amherst 01003.
Plasmid. 1994 Mar;31(2):138-47. doi: 10.1006/plas.1994.1015.
We have determined the nucleotide sequence of IS401, an insertion sequence implicated in rearrangements of a 170-kb cryptic plasmid from Pseudomonas cepacia. Our analysis focused on a 4066-bp plasmid fragment containing adjacent copies of IS401 and of IS408, an element reported previously to activate gene expression in P. cepacia. One objective was to determine if an apparent increase in the copy number of IS401 in strains carrying adjacent plasmid copies of these two elements might be due to readthrough transcription of an IS401 transposase gene from an outwardly directed promoter within IS408. This possibility was ruled out by nucleotide sequence analysis of the 4066-bp plasmid fragment, which indicated that the major open reading frames of IS401 were oriented in the direction of IS408. IS401 was 1316 bp in length and had 26-bp terminal inverted repeats flanked by 3-bp direct duplications of adjacent DNA. It was closely related to the IS3 family elements IS51 from P. savastanoi and IS3411 from Escherichia coli. Pertinent features of IS408 are also discussed.
我们已经确定了IS401的核苷酸序列,IS401是一种插入序列,与洋葱伯克霍尔德菌中一个170 kb隐蔽质粒的重排有关。我们的分析集中在一个4066 bp的质粒片段上,该片段包含IS401和IS408的相邻拷贝,IS408是先前报道的能激活洋葱伯克霍尔德菌中基因表达的元件。一个目标是确定在携带这两个元件相邻质粒拷贝的菌株中,IS401拷贝数的明显增加是否可能是由于IS401转座酶基因从IS408内的一个向外定向启动子进行通读转录所致。通过对4066 bp质粒片段的核苷酸序列分析排除了这种可能性,该分析表明IS401的主要开放阅读框是朝着IS408的方向排列的。IS401长度为1316 bp,有26 bp的末端反向重复序列,两侧是相邻DNA的3 bp直接重复序列。它与来自番茄溃疡病菌的IS51和来自大肠杆菌的IS3411等IS3家族元件密切相关。还讨论了IS408的相关特征。