Aranda-Olmedo Isabel, Tobes Raquel, Manzanera Maximino, Ramos Juan L, Marqués Silvia
Consejo Superior de Investigaciones Científicas, Estación Experimental del Zaidín, Departamento de Bioquímica y Biología Molecular y Celular de Plantas, Apdo. de correos 419, E-18080 Granada, Spain.
Nucleic Acids Res. 2002 Apr 15;30(8):1826-33. doi: 10.1093/nar/30.8.1826.
Pseudomonas putida KT2440 is a soil bacterium that effectively colonises the roots of many plants and degrades a variety of toxic aromatic compounds. Its genome has recently been sequenced. We describe that a 35 bp sequence with the structure of an imperfect palindrome, originally found repeated three times downstream of the rpoH gene terminator, is detected more than 800 times in the chromosome of this strain. The structure of this DNA segment is analogous to that of the so-called enterobacteriaceae repetitive extragenic palindromic (REP) sequences, although its sequence is different. Computer-assisted analysis of the presence and distribution of this repeated sequence in the P.putida chromosome revealed that in at least 80% of the cases the sequence is extragenic, and in 82% of the cases the distance of this extragenic element to the end of one of the neighbouring genes was <100 bp. This 35 bp element can be found either as a single element, as pairs of elements, or sometimes forming clusters of up to five elements in which they alternate orientation. PCR scanning of chromosomes from different isolates of Pseudomonas sp. strains using oligonucleotides complementary to the most conserved region of this sequence shows that it is only present in isolates of the species P.putida. For this reason we suggest that the P.putida 35 bp element is a distinctive REP sequence in P.putida. This is the first time that REP sequences have been described and characterised in a group of non-enterobacteriaceae.
恶臭假单胞菌KT2440是一种土壤细菌,能有效定殖于多种植物的根部,并降解多种有毒芳香化合物。其基因组最近已完成测序。我们发现,最初在rpoH基因终止子下游重复三次的一段35bp序列,在该菌株的染色体中被检测到800多次。该DNA片段的结构类似于所谓的肠杆菌科重复基因外回文(REP)序列,尽管其序列不同。对该重复序列在恶臭假单胞菌染色体中的存在和分布进行计算机辅助分析发现,在至少80%的情况下,该序列位于基因外,在82%的情况下,该基因外元件与相邻基因之一末端的距离小于100bp。这个35bp的元件可以作为单个元件、成对元件存在,有时还会形成多达五个元件的簇,它们的方向交替。使用与该序列最保守区域互补的寡核苷酸对不同假单胞菌菌株分离株的染色体进行PCR扫描,结果表明它仅存在于恶臭假单胞菌物种的分离株中。因此,我们认为恶臭假单胞菌的35bp元件是恶臭假单胞菌中一种独特的REP序列。这是首次在一组非肠杆菌科细菌中描述和鉴定REP序列。