Goryacheva I I, Gorelova T V, Andrianov B V
Genetika. 2015 Dec;51(12):1345-50.
Wolbachiapipientis is an obligate intracellular endosymbiont that commonly infects arthropods. Comparative genomic studies of Wolbachia reveal traces of numerous events of intergenic and intragenic recombination. The molecular mechanisms of recombination in Wolbachia are not currently known. We conducted experimental verification of the possibility of recombination of two strains of Wolbachia: wMel and wRi, after using these strains for double infection of the Dm2008Wb1 (D. melanogaster) cell culture clone permissive to Wolbachia. We obtained cell culture subclones with double Wolbachia infection and subclones infected only by strain wMel. Dual infection with the Wolbachia strains wMel and wRi has been stably maintained in the subclones for two years. Multilocus sequence typing (MLST) of the obtained subclones revealed the presence of dual infection for all five Wolbachia genes used for MLST Cloning and nucleotide sequence analysis of individual forms of the fbpA gene of Wolbachia from cell clones with dual infection showed intragenic recombination events between strains wMel and wRi, which occurred in the permanent D. melanogaster culture cell culture. The fact that putative recombination sites contain no insertions of nucleotide sequences of phages or IS elements, as well as the asymmetrical character of recombinants, favors the hypothesis that gene conversion is the most probable molecular mechanism of recombination in Wolbachia.
沃尔巴克氏体（Wolbachia pipientis）是一种专性细胞内共生菌，通常感染节肢动物。对沃尔巴克氏体的比较基因组研究揭示了许多基因间和基因内重组事件的痕迹。目前尚不清楚沃尔巴克氏体中重组的分子机制。我们在使用wMel和wRi这两种沃尔巴克氏体菌株对允许沃尔巴克氏体感染的Dm2008Wb1（黑腹果蝇）细胞培养克隆进行双重感染后，对这两种菌株重组的可能性进行了实验验证。我们获得了双重感染沃尔巴克氏体的细胞培养亚克隆以及仅被wMel菌株感染的亚克隆。沃尔巴克氏体菌株wMel和wRi的双重感染在亚克隆中稳定维持了两年。对获得的亚克隆进行多位点序列分型（MLST）显示，用于MLST的所有五个沃尔巴克氏体基因均存在双重感染。对双重感染细胞克隆中沃尔巴克氏体fbpA基因的各个形式进行克隆和核苷酸序列分析，结果显示wMel和wRi菌株之间发生了基因内重组事件，这些事件发生在黑腹果蝇的永久性培养细胞培养物中。推测的重组位点不包含噬菌体或IS元件的核苷酸序列插入，以及重组体的不对称特征，支持了基因转换是沃尔巴克氏体中最可能的重组分子机制这一假说。
