Genetic manipulation of Porphyromonas gingivalis.
Genetic manipulation of Porphyromonas gingivalis.
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DOI：
10.1002/9780471729259.mc13c02s05
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发表时间：
2007-06-01
影响因子：
--
通讯作者：
Progulske-Fox, Ann
中科院分区：
文献类型：
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作者：
Belanger, Myriam;Rodrigues, Paulo;Progulske-Fox, Ann
牙龈卟啉单胞菌是一种口腔厌氧菌，是牙周病的重要病原体，也可能导致心血管疾病、早产和糖尿病。因此，遗传学研究对于研究牙龈卟啉单胞菌毒力的分子机制至关重要。虽然分子遗传学工具已经用于许多细菌物种一段时间，但直到最近才开发出卟啉单胞菌属物种的遗传操作，并且仍然有限。在本单元中，描述了使用自杀载体pPR-UF 1和转座子Tn 4351在牙龈卟啉单胞菌中构建突变体的当前分子遗传学方法，以及进行电穿孔和缀合的方案。此外，提供了通过使用载体pT-COW的互补来恢复突变体的野生型表型的技术。最后，一个非侵入性的报告系统，允许牙龈卟啉单胞菌的基因表达和调控的研究的描述完成这个单位。
Porphyromonas gingivalis, an oral anaerobic bacterium, is an important etiological agent of periodontal disease and may contribute to cardiovascular disease, preterm birth, and diabetes as well. Therefore, genetic studies are of crucial importance in investigating molecular mechanisms of P. gingivalis virulence. Although molecular genetic tools have been available for many bacterial species for some time, genetic manipulations of Porphyromonas species were not developed until more recently and remain limited. In this unit, current molecular genetic approaches for mutant construction in P. gingivalis using the suicide vector pPR-UF1 and the transposon Tn4351 are described, as are protocols for performing electroporation and conjugation. Furthermore, a technique to restore the wild-type phenotype of the mutant by complementation using vector pT-COW is provided. Finally, a description of a noninvasive reporter system allowing the study of gene expression and regulation in P. gingivalis completes this unit.