バイオフィルム形成 Streptococcus constellatus

1 2 : 75 82. 2013 ; 76. 7606. Streptococcus constellatus .. Streptococcus constellatus S. anginosus, S. intermedius anginosus group .. Bacillus subtilis, Rothia mucilaginosa S. constellatus .. S. constellatus H39 sequencing . H39 DNA GS Junior System . DNA .. H39 2,000 .. S. constellatus H39 .. AGS .. Anginosus streptococci AGS milleri . 4. Streptococcus anginosus, Strepto- . coccus constellatus S. constellatus . Streptococcus intermedius S. intermedius H39 6 Yamanaka H39 . 3 recN, groEL, tuf, 16S ribo- somal RNA . 1, 2 3. S. constellatus 7 H39 . SEM .. 1, 2.. 1, 4 6 . 5 Costerton 8 .. 573-1121 8-1. 2 : 75 82. 2013 ; 76. PCR Purification Kit Qiagen, MD, USA . Fukushima Rapid Library Preparation Kit . Prevotella intermedia P. inter- RL . media Prevotella nigrescens P. ni- Agencourt AMPure XP Kit . grescens exopolysaccharide . EPS H39 genome library . 9, 10. 4 Emulsion polymerase chain reaction emul- Streptococcus oralis 11 Yamane sion PCR.

2 Actinomyces oris EPS H39 genome library . emulsion PCR GS Junior emPCR Kit Roche 12. H39 Diagnostics . DNA emulsion . PCR 94 . C 4 DNA 94 . C 30. 13 58 . C 4 30 68.. C 30 50 . DNA . S. constellatus H39 . 5 . Allegra .. pico titer plate 1 GS Junior Titanium PicoTiterPlate Kit, Roche Diag- 80 . C S. nostics Pico titer plate . constellatus H39 GS Junior, Roche Diagnostics . 80 . C H39 5 . yeast extract Difco, Detroit, MI, USA 6 de novo . trypticase soy agar TSA BBL Micro- GS. biology Systems, Cockeysville, MD, USA 10 de novo Assembler Software version . CO2 48 Roche Diagnostics .. 2 Genome DNA 7 . TSA H39 trypticase soy broth . BBL Microbiology Systems 10 CO2 . 24 5,000 g 10 5 L DNA 10 ng/. CR 21 L 1 L 10 M 25 L GoTaq M 2 DNA Polymerase Green Master Mix Promega . mg/mL N-acetylmuramidase 18 L MJ Mini Gradient 20 mg/mL lysozyme Wako 30 Thermal Cycler Bio-Rad Japan PCR. MagExtractor TOYOBO PCR 94 . C 2 DNA 94. genome DNA . C 1 60 . C 1 74.. C 35 74.

3 C 7 . 3 Genome library 1 TAE. H39 genome DNA Rapid Library Preparation 100 V, 20. Kit Roche Diagnostics . Rapid Library QIA quick Nebulizers MiniElute Gel Extraction Kit QIAGEN . 2 : 75 82. 2013 ; 76 S. constellatus biofilm . 1,938,626 bp 51,016 bp Sanger Table Big Dye Terminator Cycle Se- 1 . quencing Kit Applied Bio-system, CA, USA 3 . ABI PRISM 3730 DNA analyzer Applied MiGAP 3 rRNA, 45. Bio-system Sequencher tRNA, 1,926 CDSs . version , Gene Codes, MI, USA H39 hemolysin 3 sortase 6 hyaluronidase 8 1 8 . MetaGeneAnnotator pgcA Fig. 2 . version protein coding sequences . CDSs transfer RNA tRNA Table 1 Distribution of contigs assembled from messenger RNA rRNA pyrosequencing data tRNAscan-SE version Contig number Length bp . RNAmmer version 001 297,046. Microbial Genome Annotation Pipeline MiGAP 002 292,693. http : 003 171,006. 004 169,686. NCBI BLAST version . 005 99,479. 006 99,067. 007 93,289. 008 84,117. 009 82,078. 010 74,518. 1 011 64,062.

4 H39 genome library 012 52,491. 198,265 013 47,762. 014 47,376. 84,169,816 bp Fig. 1 . 015 46,222. 2 016 42,052. 017 38,847. 018 34,249. 019 27,547. 37 020 21,894. 021 12,464. 022 8,492. 023 6,799. 024 5,352. 025 4,821. 026 4,145. 027 1,445. 028 1,436. 029 1,357. 030 1,263. 031 1,056. 032 1,045. 033 922. 034 861. 035 627. 036 559. Fig. 1 Size distribution of reads of pyrosequencing 037 501. 2 : 75 82. 2013 ; 76. Fig. 4 The putative gene annotated from sequences of con- tig 009. : ABC-type polysaccharide/polyol phosphate transport sys- tem, ATPase component ;. : ABC-type polysaccharide/polyol phosphate export sys- tems, permease component ;. : glycosyltransferases involved in cell wall biogenesis ;. : lycosyltransferase ;. : glycosyltransferase ; : predicted membrane protein polysaccharide biosynthesis protein, capsular polysac- charide biosynthesis protein . Fig. 3 9 ABC-type polysaccha- ride / polyol phosphate transport system ATPase component permease component.

5 Fig. 4 .. Dental plaque .. 14.. Pseudomonas aeruginosa P. aeruginosa . Vibrio cholerae .. 1 . dental plaque . Fig. 2 Amino acid alignment of -phosphoglucomutase 8, 15, 16 . PGM sequences . Sequence 1 : PGM from Bacillus subtilis ; sequence 2 : PGM from contig 001 of strain H39 ; sequence 3 : PGM . from contig 008 of strain H39 12, 17, 18.. 16 .. Fig. 3 The putative gene annotated from sequences of con- . tig 003 . : glycosyltransferase ; : glycosyltransferase ;. : exopolysaccharide biosynthesis protein ; . : capsular exopolysaccharide family protein ; 19 H39 . : capsular polysaccharide biosynthesis protein ;.. : capsular polysaccharide biosynthesis protein . 6 Pe- 3 exopolysaccharide biosynthesis protein, tersen AGS S. intermedius . capsular exopolysaccharide family protein, capsular 20 . 2 : 75 82. 2013 ; 76 S. constellatus biofilm . S. intermedius EPS GroEL, GroES . EPS . 31 H39 . quorum-sensing system autoinducer 2. 21. Petersen quorum-sensing LuxS.

6 System competence-stimulating peptide Autoinducer 2 . S. intermedius . 20 S. intermedius H39 . AGS . Bacillus subtilis B. subtilis . pgcA. H39 yhxB 32 . glucose 6-phosphate -glucose DNA 1-phosphate -phosphoglucomutase . 3 ATP PGM . ATP . 32 34. DNA H39 . 22.. 22. H39 PGM . 6. 2 .. 6. de novo . PCR H39 2. 37 PGM . Mbp AGS Fig. 2 . S. intermedius JTH 08 2. GenBank, AP H39. 1,926 CDSs Bacillus anthracis . EPS . hemolysin 3 Fig. 3 35 . 23.. 24. sortase 6 hyaluronidase . 25.. 1 . Escherichia her- quorum-sensing system 2 mannii SEM . 3 random mutagenesis . 26 28. transposon ABC transporter wzt . P. aeruginosa EPS . quorum-sensing system 36 Yamanaka wzt . P. aeruginosa . 29.. Wzt/Wzm . 37. 30. P. nigrescens EPS H39 EPS . 2 : 75 82. 2013 ; 76. Fig. 4 Costerton JW, Cheng K-J, Geesey GG, Ladd TI, Nickel JC, Dasgupta M, Marrie TJ. Bacterial biofilms in nature ATPase . and disease. Annu Rev Microbiol 1987 ; 41 : 435 464. permease Prevotella nigrescens exopolysaccharide 1998 ; 61 : 21.

7 33.. Prevotella intermedia . 1992 ; 55 : 283 293.. 2004 ; 67 : 235 244. Yamane K, Nambu T, Yamanaka T, Ishihara K, Tatami T, Mashimo C, Walker CB, Leung K-P, Fukushima H. Pathogenicity of exopolysaccharide-producing Actinomy- ces oris isolated from an apical abscess lesion. Int Endod . J 2013 ; 46 : 145 154. Furukawa T, Yamanaka T, Yamane K, Mashimo C, Su- C . 23592724 . B . 24791975 mita Y, Sugimori C, Walker CB, Leung K-P, Fukushima H. Oral streptococci isolated from acute abscesses produce biofilm-forming materials. J Dent Res 2006 ; 85 : 2323.. Marsh PD. Dental plaque as a microbial biofilm. Caries 536 Res 2004 ; 38 : 204 211. 24 12 8 Govan JRW, Deretic V. Microbial pathogenesis in cystic fibrosis : mucoid Pseudomonas aeruginosa and Burk- holderia cepacia. Microbiol Rev 1996 ; 60 : 539 574. Hall-Stoodley L, Stoodley P. Biofilm formation and dis- Whiley RA, Beighton D, Winstanley TG, Fraser HY, persal and the transmission of human pathogens.

8 Trends Hardie JM. Streptococcus intermedius, Streptococcus Microbiol 2005 ; 13 : 7 10. constellatus , and Streptococcus anginosus the Strepto- Yamane K, Yamanaka T, Yamamoto N, Furukawa T, coccus milleri group : association with different body Fukushima H, Walker CB, Leung K-P. A novel exopoly- sites and clinical infections. J Clin Microbiol 1992 ; 30 : saccharide from a clinical isolate of Prevotella nigres- 243 244. cens : purification, chemical characterization and possible Piscitelli SC, Shwed J, Schreckenberger P, Danziger role in modifying human leukocyte phagocytosis. Oral Mi- LH. Streptococcus milleri group : renewed interest in an crobiol Immunol 2005 ; 20 : 1 9. elusive pathogen. Eur J Clin Microbiol Infect Dis 1992 ; Yamane K, Ogawa K, Yoshida M, Hayashi H, Naka- 11 : 491 498. mura T, Yamanaka T, Tamaki T, Hojoh H, Leung K-P, Ruoff KL. Streptococcus anginosus Streptococcus Fukushima H. Identification and characterization of clini- milleri : the unrecognized pathogen.

9 Clin Microbiol Rev cally isolated biofilm-forming gram-positive rods from teeth 1988 ; 1 : 102 108. associated with persistent apical periodontitis. J Endod Whiley RA, Fraser H, Hardie JM, Beighton D. Pheno- 2009 ; 35 : 347 352. typic differentiation of Streptococcus intermedius, Strepto- Fux CA, Costerton JW, Stewart PS, Stoodley P. Sur- coccus constellatus , and Streptococcus anginosus strains vival strategies of infectious biofilms. Trends Microbiol within the Streptococcus milleri group . J Clin Microbiol 2005 ; 13 : 34 40. 1990 ; 28 : 1497 1501. Petersen FC, Pecharki D, Scheie AA. Biofilm mode of Narikiyo M, Tanabe C, Yamada Y, Igaki H, Tachimori growth of Streptococcus intermedius favored by a Y, Kato H, Muto M, Montesano R, Sakamoto H, Nakajima competence-stimulating signaling peptide. J Bacteriol Y, Sasaki H. Frequent and preferential infection of Tre- 2004 ; 186 : 6327 6331. ponema denticola, Streptococcus mitis, and Streptococ- Petersen FC, Pasco S, Ogier J, Klein JP, Assev S, cus anginosus in esophageal cancers.

10 Cancer Sci 2004 ; Scheie AA. Expression and functional properties of the 95 : 569 574. Streptococcus intermedius surface protein antigen I/II. In- fect Immun 2001 ; 69 : 4647 4653. Streptococcus intermedius Ronaghi M. Pyrosequencing sheds light on DNA se- 2008 ; 71 : 131 138. quencing. Genome Res 2001 ; 11 : 3 11. Yamanaka T, Furukawa T, Yamane K, Nambu T, Marshall R, Kaufman AK. Production of deoxyribonucle- Mashimo C, Maruyama H, Inoue J, Kamei M, Yasuoka H, ase, ribonuclease, coagulase, and hemolysins by anaero- Horiike S, Leung K-P, Fukushima H. Biofilm-forming ca- bic gram-positive cocci. J Clin Microbiol 1981 ; 13 : 787 . pacity on clinically isolated Streptococcus constellatus 788. from an odontogenic subperiosteal abscess lesion. J Bac- Yamaguchi T, Matsumoto M, Sugimoto Y, Soutome S, teriol Parasitol 2013 ; in press Oho T. Gene cloning and characterization of Streptococ- 2 : 75 82. 2013 ; 76 S. constellatus biofilm . cus intermedius fimbriae involved in saliva-mediated ag- Karamata D.