Korean Journal of Environmental Biology (환경생물)

Korean Society of Environmental Biology (한국환경생물학회)
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Use of 16S-23S rRNA Intergenic Spacer Region for Rapid Detection of Vibrio fluvialis

16S-23S rRNA Intergenic Spacer Region을 이용한 Vibrio fluvialis의 검출

  • 강현실 (제주대학교 해양과학대학 해양생산과학부) ;
  • 허문수 (제주대학교 해양과학대학 해양생산과학부) ;
  • 이제희 (제주대학교 해양과학대학 해양생산과학부)
  • Published : 2003.03.01
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Abstract

We have examined the 16S-23S rRNA intergenic spacer regions (ISR) of Vibrio fluvialis. ISRs were PCR amplified, cloned into a plasmid vector and then sequenced. As results of ISR nucleotide sequence analysis, total of 6 clones were isolated depending on the size. The clones were different in both the number and the composition of the tRNA genes, and were designated ISR-A, ISR-E, ISR-El, ISR-lA, ISR-EKV, ISR-EKAV. ISR-A contains $tRNA^{Ala}$; ISR-lA, $tRNA^{Ile}$-$tRNA^{Ala}$; ISR-EKV, $tRNA^{GIu}$-$tRNA^{Lys}$-$tRNA^{Val}$;ISE-EKAV, $tRNA^{GIu}$-$tRNA^{Lys}$-$tRNA^{Ala}$-$tRNA^{Val}$; ISR -E and E1, $tRNA^{GIu}$ clusters. ISR-EKV was shown to be a minor type out of the six ISR types and showed a very limited homology between ISR-EKV from V, fluvialis and ISRa from other Vibrio species. Therefore ISR-EKV sequence was used to design species-specific primers to detect V, fiuvialis from other Vibrio species by PCR reaction. The specificity of the primers was examined using genomic DNA of other Vibrios as templates for PCR reaction. The result showed that PCR can be a useful method to detect V. fluvialis among Vibrio species in a single PCR reaction.

본 연구는 위장관염을 일으키는 Vibrio fluvialis의 16S-23S rRNA intergenic spacer region을 분석하였다. ISR을 PCR 증폭 후 plasmid vector에 클로닝하여 염기서열을 분석하였다. 그 결과, ISR의 염기서열은 tRNA gene 조성과 크기에 따라 총 6개의 type으로 분류되었다. 각 type은 tRNA gene 조성과 수에 따라 ISR-A, ISR-E, ISR-El, ISR-lA, ISR-EKV, ISR-EKAV로 명명하였으며, ISR-A는 tRN $A^{Ala}$; ISR-lA, tRN $A^{Ile}$-tRN $A^{Ala}$; ISR-EKV, tRN $A^{Glu}$-tRN $A^{Lys}$-tRN $A^{Val}$; ISR-EKAV, tRN $A^{Glu}$-tRN $A^{Lys}$-tRN $A^{Ala}$-tRN $A^{Val}$; ISR-E와 El은 tRN $A^{Glu}$를 갖고 있었다. 이 중 ISR-EKV type은 minor type으로 존재하고 있으며, 여러 Vibrio종의 ISR-EKV type과 비교시 변이성이 높은 부위를 확인하였다. 따라서, 이 ISR-EKV의 염기서열을 여러 Vibrio종에서 V. fluuialis를 검출하기 위한 species-specific primer 제작에 이용하였다. 제작된 primer의 특이성은 여러 Vibrio 의 genomic DNA를 분리하여 PCR 반응으로 확인하였다. 그 결과, 제작된 primer는 V. fluvialis에 종 특이성이 있으며 여러 Vibrio종으로부터 빠른 검출이 가능함을 확인하였다.로부터 빠른 검출이 가능함을 확인하였다.

Keywords

References

  1. 의학 생물학. 대한미생물학회편 강재승;고광균;국윤호;김각균;김경호;김경희
  2. Vibrio fluvialis에 의한 위장관염 1예. 감염 v.30 이혜정;이성욱;배영진;박진곤;최필선;조강일;천종철;문성채;황은주
  3. J. Mol. Evol/ v.47 Sequence diversity in the 16S-23S intergenic spacer region (ISR) of the rRNA operons in representatives of the Excherichia coli ECOR collection Anton AI;AJ Martinez-Murcia;F Rodriguez-Valera https://doi.org/10.1007/PL00006363
  4. Int. J. Syst. Bacteriol v.44 Sequence determination of rRNA genes of pathogenic Vibrio species and whole-cell identification of Vibrio vulnificus with rRNA-targetted oligonucleotide probes Aznar R;W Ludwig;RI Amann;KH Schleifer https://doi.org/10.1099/00207713-44-2-330
  5. J. Microbiol. Methods. v.36 Detection of total and hemolysin-producing Vibrio parahaemolyticus in shellfish using multiplex PCR amplification of tl, tdh and trh Bej AK;DP Patterson;CW Brasher;MC Vickery;DD Jones;CA Kaysner https://doi.org/10.1016/S0167-7012(99)00037-8
  6. Microbiol. Rev. v.59 Control of rRNA transcription on Escherichia Condon C;C Squires;CL Squires
  7. Appl. Environ. Microbiol v.65 Analysis of 16S-23S rRNA intergenic spacer regions of Vibrio cholerae and Vibrio mimicus Chun J;A Huq;RR Colwell
  8. Int. J. Syst. Bacteriol v.42 Towards a phylogeny of the genus Vibrio based on 16S rRNA sequences Dorsch M;D Lane;E Stackebrandt https://doi.org/10.1099/00207713-42-1-58
  9. J. Bacteriol v.178 Comparison of the small 16S to 23S intergenic spacer region (ISR) of the rRNA operons of some Excherichia coli strains of the ECOR collection and E. coli K-12 Garcia-Martinez J;A Martinez-Murcia;AI Anton;F Rodriguez-Valera
  10. J. Microbiol. Methods v.36 Use of the 16S23S ribosomal genes spacer region in studies of prokaryotic diversity Garcia-Martinez J;SG Acinas;AI Anton;F Rodriguez-Valera https://doi.org/10.1016/S0167-7012(99)00011-1
  11. Microbiol v.142 New approaches to typing and identification of bacteria using the 16S-23S rDNA spacer region Gurtler V;VA Stanisich https://doi.org/10.1099/13500872-142-1-3
  12. J. Clin. Microbiol v.20 Vibrio fluvialis and Vibrio furnissii isolated from a stool sample of one patient Hickman-Brenner FW;DJ Brenner;AG Steigerwalt;M Schreiber;SD Holmberg;LM Baldy;CS Lewis;NM Pickens;JJ Farmer III
  13. J. Clin. Microbiol v.11 Iso-lation of Vibrio like-group, EF-6, from patients with diarrhea Huq MI;AK Alam;DJ Brenner;GK Morris
  14. J. Korean Soc. Food Sci. Nutr. v.30 Distribution of extracellular proteases from various Vibrio species Kim YH;CR Jung;SK Kim;JY Yang;JH Cha
  15. Int. J. Syst. Bacteriol v.43 Phylogenetic relationship of marine bacteria, mainly members of the family Vibrionaceae, determined on the basis of 16S rRNA sequences Kita-Tsukamoto K;H Oyaizu;K Nanba;U Simidu https://doi.org/10.1099/00207713-43-1-8
  16. South. Med. J. v.90 Vibrio fluvialis;an underrecognized enteric pathogen in infants? Kolb AK;SC Eppes;JD Klein https://doi.org/10.1097/00007611-199705000-00017
  17. Microbiol. Immunol v.44 The O-polysaccharide of lipopolysaccharide iso-lated from Vibrio fluvialis O19 is identical to that Vib-rio bioserogroup 1875 variant Kondo S;Y Haishima;K Isshida;K Hisatsune
  18. Marine Pollution Bulletin v.38 Identification of oligonucleotide primers targeted at the 16S-23S rDNA intergenic spacers Genus- and species-specific detection of Aeromonads Kong RYC;A Pelling;CL So;RSS Wu https://doi.org/10.1016/S0025-326X(99)00044-2
  19. J. Appl. Bacteriol v.50 Taxomony and description of Vibrio fluvialis sp. nov. (symonym group F vibrios, Group EF6). Lee JV;P Shread;AL Furniss;TN Bryant https://doi.org/10.1111/j.1365-2672.1981.tb00873.x
  20. Infect. Immun v.35 Detection of toxins produced by Vibrio fluvialis Lockwood DE;AS Kreger;SH Richardson
  21. Nucleic Acids Res. v.25 tRNAscan-SE;a program for improved detection of transfer RNA genes in genomic sequence Lowe TM;SR Eddy https://doi.org/10.1093/nar/25.5.955
  22. FEMS Microbiol. Lett v.192 Structural variation in the 16S-23S rRNA intergenic spacers of Vibrio parahaemolyticus Maeda T; N Takada;M Furchita;T Shiba
  23. Appl. Environ. Microbiol v.60 Rapid detection and identification of Vibrio anguillarum by using a specific oligonucleotide probe complementary to 16S rRNA Martinez-Picado J;AR Blanch;J Jofre
  24. Appl. Environ. Microbiol v.45 Medium-dependent production of extracellular enterotoxins by non-o-1 Vibrio cholerae, Vibrio mimicus, and Vibrio fluvialis Nishibuchi M;RJ Seidler
  25. Appl. Environ. Microbiol v.46 Broth medium for enrichment of Vibrio fluvialis from the the environment Nishibuchi M;NC Roberts;HB Jr Bradford;RJ Seidler
  26. J. Clin. Microbiol v.23 Feasibility of sameday identification of membes of the family Vibrionaceae by the API 20E system Overman TL;JK Overley
  27. J. Diarrhoeal Dis. Res. v.14 Factors affecting production of haemolysin by strains of Vibrio fluvialis Rahim Z;KM Aziz
  28. Microbiol Immunol v.36 Lipopolysaccharide composition and virulence properties of clinical and environmental strains in Vibrio fluvialis and Vibrio mimicus Rahman MM;F Qadri;MJ Albert;A Hossain;M Mosihuzzaman
  29. Appl. Envinron. Microbiol. v.55 Identification of Vibrio anguillarum in fish using the partial 16S rRNA sequences and specific 16S rRNA oligonucleotide probe Rehnstam AS;A Norqvust;H Wolf-Wata;A Hagstrom
  30. Ann. Rev. Microbiol. v.44 Mechanism and regulation of bacterial RNA processing Srivastava AK;D Schlessinger https://doi.org/10.1146/annurev.mi.44.100190.000541
  31. Infect. Immun v.46 Production and partial characterization of Vibrio fluvialis cytotoxin Wall VW;AS Kreger;SH Richardson
  32. Microbiol Rev. v.49 Structure and organization of genes for transfer rilbonucleic acid in Bacillus subtilis Vlod BS