Culturing and Genetically Manipulating Entomopathogenic Nematodes

Christa Heryanto; Ramesh Ratnappan; Damien M. O'Halloran; John M. Hawdon; Ioannis Eleftherianos

doi:10.3791/63885

JoVE Journal
Immunology and Infection

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JoVE Journal Immunology and Infection

Culturing and Genetically Manipulating Entomopathogenic Nematodes

培养和遗传操纵昆虫致病线虫

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07:17 min

March 31, 2022

DOI: 10.3791/63885-v

Christa Heryanto¹, Ramesh Ratnappan², Damien M. O'Halloran¹, John M. Hawdon², Ioannis Eleftherianos¹

¹Department of Biological Sciences,The George Washington University, ²Department of Microbiology, Immunology, and Tropical Medicine, School of Medicine and Health Sciences,The George Washington University

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昆虫致病线虫与细菌共生，它们通过破坏其先天免疫系统成功地感染昆虫。为了促进线虫感染的遗传基础研究，描述了维持和遗传操纵昆虫致病线虫的方法。

维持和扩增致虫病理线虫的有效方案对于了解线虫致病性和宿主抗线虫免疫的分子基础至关重要。该协议使我们能够使用昆虫宿主梅洛内拉菌产生大量共生和轴性异性异性结节性细菌和碳甲壳类斯坦纳菌IJ。首先用一张滤纸覆盖培养皿，并加入大约10至15个梅氏菌幼虫。

使用移液管，将每10微升悬浮液中含有约25至50个感染性幼体的两毫升水分配到蜡虫上。并将培养皿在室温下存放在柜中。根据滤纸的水分，每两天加入一到两毫升水。

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