van der Zee A, Agterberg C, van Agterveld M, Peeters M, Mooi F R
Laboratory of Medical Microbiology, St. Elisabeth Hospital, Tilburg, The Netherlands.
J Bacteriol. 1993 Jan;175(1):141-7. doi: 10.1128/jb.175.1.141-147.1993.
By analysis of repetitive DNA in Bordetella parapertussis, an insertion sequence element, designated IS1001, was identified. Sequence analysis revealed that IS1001 comprised 1,306 bp and contained inverted repeats at its termini. Furthermore, several open reading frames that may code for transposition functions were identified. The largest open reading frame coded for a protein comprising 406 amino acid residues and showed homology to TnpA, which is encoded by an insertion sequence element (IS1096) found in Mycobacterium smegmatis. Examination of flanking sequences revealed that insertion of IS1001 occurs preferentially in stretches of T's or A's and results in a duplication of target sequences of 6 to 8 bases. IS1001 was found in about 20 copies in 10 B. parapertussis strains analyzed. No restriction fragment length polymorphism was observed in B. parapertussis when IS1001 was used as a probe. An insertion sequence element similar or identical to IS1001 was found in B. bronchiseptica strains isolated from pigs and a rabbit. In these strains, about five copies of the IS1001-like element were present at different positions in the bacterial chromosome. Neither B. pertussis nor B. bronchiseptica strains isolated from humans and dogs contained an IS1001-like element. Therefore, IS1001 may be used as a specific probe for the detection of B. parapertussis in human clinical samples.
通过对副百日咳博德特氏菌中重复DNA的分析,鉴定出一种插入序列元件,命名为IS1001。序列分析表明,IS1001由1306个碱基对组成,其末端含有反向重复序列。此外,还鉴定出几个可能编码转座功能的开放阅读框。最大的开放阅读框编码一种由406个氨基酸残基组成的蛋白质,与耻垢分枝杆菌中发现的插入序列元件(IS1096)编码的TnpA具有同源性。对侧翼序列的检查表明,IS1001优先插入T或A的序列中,并导致6至8个碱基的靶序列重复。在所分析的10株副百日咳博德特氏菌菌株中,发现IS1001约有20个拷贝。当使用IS1001作为探针时,在副百日咳博德特氏菌中未观察到限制性片段长度多态性。在从猪和兔子分离的支气管败血博德特氏菌菌株中发现了与IS1001相似或相同的插入序列元件。在这些菌株中,细菌染色体的不同位置存在约五个拷贝的IS1001样元件。从人和狗分离的百日咳博德特氏菌和支气管败血博德特氏菌菌株均未含有IS1001样元件。因此,IS1001可作为检测人类临床样本中副百日咳博德特氏菌的特异性探针。