Insects are frequently infected by bacterial symbionts that greatly affect their physiology and ecology. Most of these endosymbionts are, however, barely tractable outside their native host, rendering functional genetics studies difficult or impossible. Spiroplasma poulsonii is a facultative bacterial endosymbiont of Drosophila melanogaster that manipulates the reproduction of its host by killing its male progeny at the embryonic stage. S. poulsonii, although a very fastidious bacterium, is closely related to pathogenic Spiroplasma species that are cultivable and genetically modifiable. In this work, we present the transformation of S. poulsonii with a plasmid bearing a fluorescence cassette, leveraging techniques adapted from those used to modify the pathogenic species Spiroplasma citri. We demonstrate the feasibility of S. poulsonii transformation and discuss approaches for mutant selection and fly colonization, which are persisting hurdles that must be overcome to allow functional bacterial genetics studies of this endosymbiont in vivo.

中文翻译：

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果蝇性别操纵性内共生螺旋体poulsonii和持久障碍的转化为功能遗传研究。

昆虫经常被细菌共生菌感染，这极大地影响了它们的生理和生态。然而，大多数这些共生共生体在它们的天然宿主外几乎很难处理，这使得功能遗传学研究困难或不可能。螺螺旋体是果蝇的兼性细菌内共生体，通过杀死胚胎的雄性后代来操纵宿主的繁殖。尽管Sul poulsonii是一种非常挑剔的细菌，但它与可培养和遗传修饰的致病性螺旋体物种密切相关。在这项工作中，我们介绍了S. poulsonii的转化利用带有荧光盒的质粒，利用从用于修饰病原性物种柠檬螺旋体的技术改编的技术。我们证明了S. poulsonii转化的可行性，并讨论了突变选择和果蝇定植的方法，这是必须克服的持续障碍，以便在体内对该内共生体进行功能细菌遗传学研究。