Wasser Daniel, Borst Andreas, Hammelmann Mathias, Ludt Katharina, Soppa Jörg
Institute for Molecular Biosciences, Biocentre, Goethe-University, Frankfurt, Germany.
Front Microbiol. 2021 Jun 10;12:680854. doi: 10.3389/fmicb.2021.680854. eCollection 2021.
Gene conversion is defined as the non-reciprocal transfer of genetic information from one site to a homologous, but not identical site of the genome. In prokaryotes, gene conversion can increase the variance of sequences, like in antigenic variation, but can also lead to a homogenization of sequences, like in the concerted evolution of multigene families. In contrast to these intramolecular mechanisms, the intermolecular gene conversion in polyploid prokaryotes, which leads to the equalization of the multiple genome copies, has hardly been studied. We have previously shown the intermolecular gene conversion in halophilic and methanogenic archaea is so efficient that it can be studied without selecting for conversion events. Here, we have established an approach to characterize unselected intermolecular gene conversion in making use of two genes that encode enzymes involved in carotenoid biosynthesis. Heterozygous strains were generated by protoplast fusion, and gene conversion was quantified by phenotype analysis or/and PCR. It was verified that unselected gene conversion is extremely efficient and it was shown that gene conversion tracts are much longer than in antigenic variation or concerted evolution in bacteria. Two sites were nearly always co-converted when they were 600 bp apart, and more than 30% co-conversion even occurred when two sites were 5 kbp apart. The gene conversion frequency was independent from the extent of genome differences, and even a one nucleotide difference triggered conversion.
基因转换被定义为遗传信息从基因组的一个位点非相互性地转移到一个同源但不相同的位点。在原核生物中,基因转换可以增加序列的变异性,如在抗原变异中,但也可以导致序列的同质化,如在多基因家族的协同进化中。与这些分子内机制不同,多倍体原核生物中的分子间基因转换会导致多个基因组拷贝的均等化,对此几乎没有研究。我们之前已经表明,嗜盐古菌和产甲烷古菌中的分子间基因转换非常高效,以至于无需选择转换事件就能对其进行研究。在这里,我们利用两个编码类胡萝卜素生物合成相关酶的基因,建立了一种方法来表征未选择的分子间基因转换。通过原生质体融合产生杂合菌株,并通过表型分析或/和聚合酶链反应对基因转换进行定量。结果证实,未选择的基因转换极其高效,并且表明基因转换片段比细菌中的抗原变异或协同进化中的片段长得多。当两个位点相距600碱基对时,它们几乎总是同时被转换,当两个位点相距5千碱基对时,甚至会发生超过30%的同时转换。基因转换频率与基因组差异程度无关,即使只有一个核苷酸差异也会引发转换。