School of Environmental Sciences, Jawaharlal Nehru University, New Delhi, India.
PLoS One. 2013 Sep 30;8(9):e74465. doi: 10.1371/journal.pone.0074465. eCollection 2013.
Homologous recombination (HR) has not been demonstrated in the parasitic protists Entamoeba histolytica or Entamoeba invadens, as no convenient method is available to measure it. However, HR must exist to ensure genome integrity, and possible genetic exchange, especially during stage conversion from trophozoite to cyst. Here we show the up regulation of mitotic and meiotic HR genes in Entamoeba during serum starvation, and encystation. To directly demonstrate HR we use a simple PCR-based method involving inverted repeats, which gives a reliable read out, as the recombination junctions can be determined by sequencing the amplicons. Using this read out, we demonstrate enhanced HR under growth stress in E. histolytica, and during encystation in E. invadens. We also demonstrate recombination between chromosomal inverted repeats. This is the first experimental demonstration of HR in Entamoeba and will help future investigations into this process, and to explore the possibility of meiosis in Entamoeba.
同源重组(HR)尚未在寄生原生动物溶组织内阿米巴或侵袭性内阿米巴中得到证明,因为没有方便的方法来测量它。然而,HR 必须存在以确保基因组完整性和可能的遗传交换,特别是在从滋养体到囊胞的阶段转换期间。在这里,我们显示在血清饥饿和囊胞形成期间,阿米巴中的有丝分裂和减数分裂 HR 基因的上调。为了直接证明 HR,我们使用一种简单的基于 PCR 的方法,涉及反向重复序列,该方法给出了可靠的读数,因为通过对扩增子进行测序可以确定重组连接点。使用这种读数,我们证明了在 E. histolytica 的生长应激下和在 E. invadens 的囊胞形成过程中增强了 HR。我们还证明了染色体反向重复之间的重组。这是在阿米巴中首次对 HR 的实验证明,将有助于对该过程的未来研究,并探索阿米巴减数分裂的可能性。