This book addresses the major periodontal pathogens implicated as causal agents in periodontal disease, including
spp. Beginning with methods for bacterial genetic manipulation, the volume continues with sections on experimental methods to examine virulence factors, interactions with other pathogenic microorganism and host cells, as well as a chapter on an animal model of periodontitis. Written for the highly successful
series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and practical,
Periodontal Pathogens: Methods and Protocols serves as an ext
Table of ContentsPart I: Methods for Bacterial Genetic Manipulation
1. Site-Directed and Random Mutagenesis in Porphyromonas gingivalis: Construction of Fimbriae-Related-Gene Mutant
So-ichiro Nishiyama, Yoshiaki Hasegawa, and Keiji Nagano
2. Genetic Manipulations of Oral Spirochete Treponema denticola
Kurni Kurniyati and Chunhao Li
3. Construction of a Gene-Deletion Mutant in Tannerella forsythia
Keiji Nagano and Yoshiaki Hasegawa
4. Construction of a Mutant in Prevotella melaninogenica Using the Conjugation Transfer Method with Escherichia coli
Yoshio Kondo
5. Genetic Transformation of Fusobacterium nucleatum
Akihiro Yoshida and Akihiko Ikegami
Part II: Experimental Methods to Examine Virulence Factors
6. Genotyping of Porphyromonas gingivalis in Relationship to Virulence
Atsuo Amano, Youn-Hee Choi, and Hiroki Takeuchi
7. Transport and Polymerization of Porphyromonas gingivalis Type V Pili
Mikio Shoji, Satoshi Shibata, Mariko Naito, and Koji Nakayama
8. Purification of Native Mfa1 Fimbriae from Porphyromonas gingivalis
Yoshiaki Hasegawa, Keiji Nagano, Yukitaka Murakami, and Richard J. Lamont
9. Crystallization of Recombinant Fimbrial Proteins of Porphyromonas gingivalis
Thomas Heidler and Karina Persson
10. Enzymatic Characteristics and Activities of Gingipains from Porphyromonas gingivalis
Tomoko Kadowaki
11. Structural Characterization of the Type IX Secretion System in Porphyromonas gingivalis
Dhana G. Gorasia, Eric Hanssen, Paul D. Veith, and Eric C. Reynolds
12. Methods for Functional Characterization of the Type IX Secretion System of Porphyromonas gingivalis
Keiko Sato
13. Purification of Tannerella forsythia Surface (S-) Layer Proteins
Sreedevi Chinthamani, Prasad R. Settem, Kiyonobu Honma, Takuma Nakajima, and Ashu Sharma
14. Separation of Glycosylated OmpA-Like Proteins from Porphyromonas gingivalis and Tannerella forsythia
Yukitaka Murakami, Keiji Nagano, and Yoshiaki Hasegawa
15. Intranasal Vaccine Study Using Porphyromonas gingivalis Membrane Vesicles: Isolation Method and Application to a Mouse Model
Satoru Hirayama and Ryoma Nakao
16. Analysis of the Butyrate-Producing Pathway in Porphyromonas gingivalis
Yasuo Yoshida
17. Characterization of the Treponema denticola Virulence Factor Dentilisin
Yuichiro Kikuchi and Kazuyuki Ishihara
18. Evaluation of the Virulence of Aggregatibacter actinomycetemcomitans through the Analysis of Leukotoxin
Toshiyuki Nagasawa, Satsuki Kato, and Yasushi Furuichi
19. Lipoprotein Extraction from Microbial Membrane and Lipoprotein/Lipopeptide Transfection into Mammalian Cells
Akira Hasebe, Ayumi Saeki, and Ken-ichiro Shibata
Part III: Interactions with Other Pathogenic Microorganism and Host Cells
20. Analysis of the Interaction between HIV and Periodontopathic Bacteria that Reactivates HIV Replication in Latently Infected Cells
Kenichi Imai
21. Invasion of Gingival Epithelial Cells by Porphyromonas gingivalis
Hiroki Takeuchi and Atsuo Amano
22. Analysis of Interaction between Porphyromonas gingivalis and Endothelial Cells In Vitro
Kenji Matsushita
Part IV: Animal Model of Periodontitis
23. Analysis of Experimental Ligature-Induced Periodontitis Model in Mice
Hikaru Tamura, Tomoki Maekawa, Takumi Hiyoshi, and Yutaka Terao
