Department of Pharmacy, University of Tromsø, N-9037, Norway.
Genetics. 2009 Aug;182(4):1165-81. doi: 10.1534/genetics.109.103127. Epub 2009 May 27.
Naturally transformable bacteria acquire chromosomal DNA from related species at lower frequencies than from cognate DNA sources. To determine how genome location affects heterogamic transformation in bacteria, we inserted an nptI marker into random chromosome locations in 19 different strains of the Acinetobacter genus (>24% divergent at the mutS/trpE loci). DNA from a total of 95 nptI-tagged isolates was used to transform the recipient Acinetobacter baylyi strain ADP1. A total of >1300 transformation assays revealed that at least one nptI-tagged isolate for each of the strains/species tested resulted in detectable integration of the nptI marker into the ADP1 genome. Transformation frequencies varied up to approximately 10,000-fold among independent nptI insertions within a strain. The location and local sequence divergence of the nptI flanking regions were determined in the transformants. Heterogamic transformation depended on RecA and was hampered by DNA mismatch repair. Our studies suggest that single-locus-based studies, and inference of transfer frequencies from general estimates of genomic sequence divergence, is insufficient to predict the recombination potential of chromosomal DNA fragments between more divergent genomes. Interspecies differences in overall gene content, and conflicts in local gene organization and synteny are likely important determinants of the genomewide variation in recombination rates between bacterial species.
自然转化细菌从相关物种中获得染色体 DNA 的频率低于从同源 DNA 来源获得的频率。为了确定基因组位置如何影响细菌中的异源转化,我们将 nptI 标记插入到不动杆菌属的 19 个不同菌株的随机染色体位置(在 mutS/trpE 基因座处差异大于 24%)。总共使用来自 95 个 nptI 标记的分离物的 DNA 转化受体不动杆菌属 ADP1 菌株。总共进行了>1300 次转化测定,结果表明在所测试的每个菌株/物种中,至少有一个 nptI 标记的分离物导致 nptI 标记可检测地整合到 ADP1 基因组中。在一个菌株内的独立 nptI 插入之间,转化频率变化高达约 10000 倍。在转化体中确定了 nptI 侧翼区域的位置和局部序列差异。异源转化取决于 RecA,并且受到 DNA 错配修复的阻碍。我们的研究表明,基于单基因座的研究以及从基因组序列差异的一般估计推断转移频率不足以预测更具差异性基因组之间染色体 DNA 片段的重组潜力。物种间的总体基因含量差异,以及局部基因组织和同线性的冲突,可能是细菌种间重组率的全基因组变异的重要决定因素。