Riley Sean P, Macaluso Kevin R, Martinez Juan J
Vector-Borne Disease Laboratories, Department of Pathobiological Sciences, Louisiana State University School of Veterinary Medicine, Baton Rouge, Louisiana.
Curr Protoc Microbiol. 2015 Nov 3;39:3A.6.1-3A.6.20. doi: 10.1002/9780471729259.mc03a06s39.
Genetic manipulation of obligate intracellular bacteria of the genus Rickettsia is currently undergoing a rapid period of change. The development of viable genetic tools, including replicative plasmids, transposons, homologous recombination, fluorescent protein-encoding genes, and antibiotic selectable markers has provided the impetus for future research development. This unit is designed to coalesce the basic methods pertaining to creation of genetically modified Rickettsia. The unit describes a series of methods, from inserting exogenous DNA into Rickettsia to the final isolation of genetically modified bacterial clones. Researchers working towards genetic manipulation of Rickettsia or similar obligate intracellular bacteria will find these protocols to be a valuable reference.
立克次氏体属专性细胞内细菌的基因操作目前正处于快速变革时期。包括复制性质粒、转座子、同源重组、荧光蛋白编码基因和抗生素选择标记在内的可行基因工具的开发为未来的研究发展提供了动力。本单元旨在整合与创建转基因立克次氏体相关的基本方法。该单元描述了一系列方法，从将外源DNA插入立克次氏体到最终分离转基因细菌克隆。致力于立克次氏体或类似专性细胞内细菌基因操作的研究人员会发现这些方案是有价值的参考。
