Nisen P, Purucker M, Shapiro L
J Bacteriol. 1979 Nov;140(2):588-96. doi: 10.1128/jb.140.2.588-596.1979.
Plasmid and phage deoxyribonucleic acid (DNA) harboring bacterial insertion sequence (IS) elements IS1, IS2, and IS5 were characterized and used as probes to detect homologous sequences in various procaryotic and eucaryotic genomes. The hybridization method used permits the detection of sequences partially homologous to the elements. Hybridization of the IS-containing probes to each other revealed a region of limited homology between IS1 and IS2. Homologous sequences were then detected by computer analysis of the published IS1 and IS2 nucleotide sequences. The homologous sequence contains a tandemly repeated tetranucleotide sequence which resembles the repeated sequence at the hot spot for spontaneous mutations in the lacI gene (P. J. Farabaugh, U. Schmeissner, M. Hofer, and J. Miller, J. Mol. Biol. 126:847-863, 1978). Homology between the IS elements and various genomes was determined by hybridizing labeled DNA containing IS1, IS2, and IS5 sequences to Southern blots of chromosomal DNA cleaved with restriction endonucleases. IS1 and IS5 appear limited to the enteric bacteria, whereas IS2 sequences can also be detected in Pseudomonas putida, Pseudomonas aeruginosa, and Serratia marcescens. Bacteria which appear not to possess extrachromosomal elements, e.g., Caulobacter crescentus, did not show homology with any insertion sequences tested. In addition, sequences homologous to IS1, IS2, or IS5 were not detected in Saccharomyces cerevisiae, Dictyostelium discoideum, or calf thymus DNA.
携带细菌插入序列(IS)元件IS1、IS2和IS5的质粒和噬菌体脱氧核糖核酸(DNA)经过了特性鉴定,并用作探针来检测各种原核和真核基因组中的同源序列。所使用的杂交方法能够检测与这些元件部分同源的序列。含IS的探针之间的杂交揭示了IS1和IS2之间有限的同源区域。然后通过对已发表的IS1和IS2核苷酸序列进行计算机分析来检测同源序列。该同源序列包含一个串联重复的四核苷酸序列,它类似于lacI基因自发突变热点处的重复序列(P. J. 法拉博、U. 施迈斯纳、M. 霍费尔和J. 米勒,《分子生物学杂志》126:847 - 863,1978年)。通过将含有IS1、IS2和IS5序列的标记DNA与经限制性内切酶切割的染色体DNA的Southern印迹杂交,确定了IS元件与各种基因组之间的同源性。IS1和IS5似乎仅限于肠道细菌,而IS2序列在恶臭假单胞菌、铜绿假单胞菌和粘质沙雷氏菌中也能检测到。似乎不具有染色体外元件的细菌,例如新月柄杆菌,与所测试的任何插入序列都没有显示出同源性。此外,在酿酒酵母、盘基网柄菌或小牛胸腺DNA中未检测到与IS1、IS2或IS5同源的序列。