Genetic manipulation of Porphyromonas gingivalis.
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.
Bélanger
Myriam
M
University of Florida, Gainesville, Florida, USA.
Rodrigues
Paulo
P
Progulske-Fox
Ann
A
D003227:Conjugation, Genetic; D004251:DNA Transposable Elements; D015964:Gene Expression Regulation, Bacterial; D005816:Genetic Complementation Test; D005821:Genetic Techniques; D006801:Humans; D016296:Mutagenesis; D016966:Porphyromonas gingivalis / Q000235:genetics* / Q000378:metabolism; D014170:Transformation, Genetic
D004251:DNA Transposable Elements