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Use of 16S-23S rRNA Intergenic Spacer Region for Species-specific Primer Developed of Vibrio Ichthyoenteri  

Moon Young-Gun (Faculty of Applied Marine Science, Cheju National University)
Heo Moon-Soo (Faculty of Applied Marine Science, Cheju National University)
Publication Information
Korean Journal of Microbiology / v.41, no.2, 2005 , pp. 117-124 More about this Journal
Abstract
Two bacterial isolates obtained from rotifer and diseased olive flounder larvae, Paralichthys olivaceus, were identified as Vibrio ichthyoenteri based on the results of phenotypic characterization. In an attempt to develop rapid PCR method for the detection of V. ichthyoenteri, we examined the 16S-23S rRNA intergenic spacer region(ISR) of V. ichthyoenteri and developed species-specific primer for V. ichthyoenteri. Analysis of the ISR sequences showed that V. ichthyoenteri contains one type of polymorphic ISRs. The size of ISRs was 348 bp length and did not contain tRNA genes. Mutiple alignment of representative sequences from different V. species revealed several domains of high sequence variability, and allowed to design species-specific primer for detection of V. ichthyoenteri. The specificity of the primer was examined using genomic DNA prepared from 19 different V. species, isolated 18group Vibrio species and most similar sequence of other known Vibrio species. The results showed that the PCR reaction using species-specific primer designed in this study can be used to detect V. ichthyoenteri.
Keywords
bacterial enteritis; species-specific primer; tRNA gene; Vibrio ichthyoenteri; 16S-23S rRNA ISR;
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1 박성우, 오명주(2001). 어류질병. 진솔., pp.100-104
2 진창남, 이창훈, 오상필, 나오수, 허문수(2003). 양식넙치, Paralichthys olivaceus 치어의 스쿠티카충 감염경로. 한국어병학회지, 16(1), 13-21
3 G.A. Bisbal., and D.A. Bengston. 1995. Development of the digestive tract in larval summer flouder. J. Fish Biol. 47, 277-291   DOI   ScienceOn
4 Ishimaru, K., M. Akagawa-Matsushita, and K. Muroga. 1996. Vibrio ichthyoenteri sp. nov.,a pathogen of Japanese flounder Paralichthys olivaceus larvae. Inter. J.syst bacteriol. 46(1), 155-159   DOI
5 Maeda, T., N. Takada, M. Furushita and T. Shiba. 2000. Structural variation in the 16S-23S rRNA intergenic spacers of Vibrio parahaemolyticus. FEMS Microbiol. Lett. 192, 73-77   PUBMED
6 Young, R.A., R. Macklis, and J. A. Steitz. 1979. Sequence of the 16S-23S spacer region-in two ribosomal RNA operons of Escherichia coli. J. Biol. Chem 254, 3264-3271   PUBMED
7 Muroga, K., H. Yasunobu, N. Okada, & K. Masumura. 1990. Bacterial enteritis of cultured flounder Paralichthys olivaceus larvae. Dis. Aquat. Org. 9, 121-125   DOI
8 Lee HK., SS Lee. 1997. Identification of the Vibrios isolation from the Shrmp(Carngon affinis) in estuary of Nakdong river in Kor. J. Microbiol. 32, 523-537
9 이정백, 노섬, 송춘복(1995). 넙치, Paralichthys olivaceus 자어에서 분리한 장관백탁증의 원인균인 Vibrio sp.(INFL group)의 생물학적 및 생화학적 특성. 한국어병학회지, 8(2), 99-109
10 Muroga, K. 2001. Viral and bacterial diseases in larval and juvenile marine fish and shellfish : A review. Fish Pathol. 30, 71-85
11 Yasuda, K., and N. Taga. 1980. A mass culture method for Artemia salina using bacteria as food. Mer. 18, 53-62
12 Murata, O. 1987. Infectious intestinal necrosis in flounder. Fish Pathol. 22, 59-61
13 MacFaddin, J.F. 2000. Individual biochemical tests. In: Biochemical Tests for Identification of Medical Bacteria, 3rd ed. pp.1-456
14 심두생, 이주석, 허문수, 김진우(1995). 양식생물 질병진 단 연구(분리균주에 대한 혈청학적 진단 및 최소발육저 지농도 조사). 수진사업보고서, 405-423
15 Valle, L.D., L. Zanella, P. Belvedere, and L. Colombo. 2002. Use of random amplication to develop a PCR detection method for the causative agent of fish pasteurellosis, Photobacterium damselaesubsp. piscicda(Vibrionaceae). Aquaculture. 207, 187-202   DOI   ScienceOn
16  Masumura, K., Y. Iida, T. Nakai, T. Mekuchi. 1989. The effects of water temperature and fish age on a herpesvirus infection of Japanese flounder larvae, Paralichthys olivaceus. Fish Pathol. 24, 111-114   DOI
17 Yoshimizu, M., I. Kaori, K. Kazuko, I. Nao, and Takahisa. 1999. Bacteria flora of hatchery reared Japanese flounder. Bull. Fac. Fish. Hokkaido Univ. 50, 193-200
18 D-H Kim, H-J Han, S-M Kim, D-C Lee and S-I Park. 2004. Bacterial enteritis and development of the larval digestive tract in olive flounder, Paralichthys olivaceus(Temmminck & schlegel). J. Fish Diseases. 27, 497-505   DOI   ScienceOn