幽门螺杆菌中DNA转化所必需的ComE3同源物的特性分析
Characterization of a ComE3 homologue essential for DNA transformation in Helicobacter pylori.
作者信息
Yeh Yu-Ching, Lin Tzu-Lung, Chang Kai-Chih, Wang Jin-Town
机构信息
Graduate Institute of Microbiology, National Taiwan University College of Medicine, Taipei, Taiwan.
出版信息
Infect Immun. 2003 Sep;71(9):5427-31. doi: 10.1128/IAI.71.9.5427-5431.2003.
To find genes involved in natural competence in Helicobacter pylori, we used a bioinformatics database search and found two transformation-related open reading frames (ORFs): a comE3 homologue (HP1361 ORF) of Bacillus subtilis and a comL homologue (HP1378 ORF) of Neisseria gonorrhoeae. We failed to obtain an HP1378 ORF knockout mutant, while an HP1361 ORF knockout mutant was obtained by transposon shuttle mutagenesis. The DNA transformation abilities of both natural transformation and electroporation were severely impaired (frequency, <10(-9)) in the HP1361(-) mutant. Complementation with a pHel2 vector carrying the HP1361 ORF restored the capabilities of natural competence (to a frequency of 4.21 x 10(-7)) and electroporation (to 3.62 x 10(-7)). The HP1361(-) mutant showed impairment in DNA binding and uptake. The results suggest that HP1361 is a comE3 homologue and is required for DNA binding and uptake during DNA transformation.
为了寻找幽门螺杆菌中与自然感受态相关的基因，我们利用生物信息学数据库进行搜索，发现了两个与转化相关的开放阅读框（ORF）：枯草芽孢杆菌的comE3同源物（HP1361 ORF）和淋病奈瑟菌的comL同源物（HP1378 ORF）。我们未能获得HP1378 ORF敲除突变体，而通过转座子穿梭诱变获得了HP1361 ORF敲除突变体。在HP1361（-）突变体中，自然转化和电穿孔的DNA转化能力均严重受损（频率，<10^(-9)）。用携带HP1361 ORF的pHel2载体进行互补恢复了自然感受态（频率为4.21×10^(-7)）和电穿孔（频率为3.62×10^(-7)）的能力。HP1361（-）突变体在DNA结合和摄取方面表现出缺陷。结果表明，HP1361是comE3同源物，是DNA转化过程中DNA结合和摄取所必需的。
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