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Characterization of a ComE3 Homologue Essential for DNA Transformation in Helicobacter pylori

机译:幽门螺杆菌DNA转化必需的ComE3同源物的表征

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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 × 10?7) and electroporation (to 3.62 × 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.
机译:为了在幽门螺杆菌中找到与自然能力有关的基因,我们使用了生物信息学数据库搜索,找到了两个与转化相关的开放阅读框(ORF):一个 comE3 同源物(HP1361 ORF )(枯草芽孢杆菌)和淋病奈瑟氏菌 comL 同源物(HP1378 ORF)。我们未能获得HP1378 ORF基因敲除突变体,而HP1361 ORF基因敲除突变体是通过转座子穿梭诱变获得的。 HP1361 ?突变体的自然转化和电穿孔的DNA转化能力都受到严重损害(频率,<10 ?9 )。与带有HP1361 ORF的pHel2载体互补,恢复了自然能力(频率为4.21×10 ?7 )和电穿孔(至3.62×10 ?7 )的能力。 。 HP1361 ?突变体显示DNA结合和吸收受到损害。结果表明HP1361是一个 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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