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通过胚胎显微注射进行沃尔巴克氏体转染。

Wolbachia Transinfection Via Embryonic Microinjection.

机构信息

Department of Microbiology and Molecular Genetics, Michigan State University, East Lansing, MI, USA.

出版信息

Methods Mol Biol. 2024;2739:175-188. doi: 10.1007/978-1-0716-3553-7_11.

Abstract

The process of transferring Wolbachia from one species to another to establish a stable, maternally inherited infection in the target species is known as transinfection. The success of transinfection is primarily achieved through embryonic microinjection, which is the most direct and efficient means of delivering Wolbachia into the germline of the target species and establishing stable maternal transmission. For the fundamental studies, transinfection is often used to characterize Wolbachia-host interactions, including Wolbachia host range, the role of host or bacterial factors in symbiosis, and evolution of Wolbachia-host associations. For the applied studies, use of transinfection to generate a novel infection in the target species is the first step to build the weapon for both population replacement and population suppression for controlling insect pests or their transmitted diseases. For the primary dengue vector Aedes aegypti and Anopheles vectors of malaria, which either do not naturally carry Wolbachia or are infected with strains that lack necessary features for implementation, transinfection can be established by introducing a novel strain capable of inducing both cytoplasmic incompatibility (CI) and pathogen blocking. For A. albopictus and Culex mosquito species, which naturally harbor CI-inducing Wolbachia, transinfection can be achieved by either introducing a novel strain to generate superinfection or replacing the native infection with a different Wolbachia strain in a symbiont-free line, which is derived from antibiotic treatment of the wild type. Here, we use A. aegypti as an example to describe the Wolbachia transinfection method, which can be adapted to other insect species, such as planthoppers, according to their specific developmental requirements.

摘要

将沃尔巴克氏体从一个物种转移到另一个物种,以在目标物种中建立稳定的母系遗传感染的过程称为转染。转染的成功主要通过胚胎显微注射来实现,这是将沃尔巴克氏体递送到目标物种的生殖系并建立稳定的母系传递的最直接和有效的方法。对于基础研究,转染常用于表征沃尔巴克氏体-宿主相互作用,包括沃尔巴克氏体的宿主范围、宿主或细菌因素在共生中的作用以及沃尔巴克氏体-宿主关联的进化。对于应用研究,在目标物种中产生新的感染是利用转染来构建用于控制害虫或其传播疾病的种群替换和种群抑制的武器的第一步。对于主要的登革热媒介埃及伊蚊和疟疾的疟蚊媒介,它们要么自然携带沃尔巴克氏体,要么感染的菌株缺乏实施所必需的特征,因此可以通过引入一种能够同时诱导细胞质不亲和(CI)和病原体阻断的新菌株来建立转染。对于自然携带 CI 诱导沃尔巴克氏体的白纹伊蚊和库蚊物种,可以通过引入新菌株来产生超感染,或通过在无共生体的系中用不同的沃尔巴克氏体菌株替换天然感染来实现转染,该系是通过对野生型进行抗生素处理衍生而来的。在这里,我们以埃及伊蚊为例来描述沃尔巴克氏体转染方法,根据其特定的发育要求,可以将该方法适应于其他昆虫物种,例如粉虱。

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