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제주도 Bovine Rotavirus의 분리와 그 특성
유제현,이영건,주지선,김유성,김순재,T Ebina,O Nakagomi,S Urasawa,K Taniguchi,Herry B . Greenberg 대한바이러스학회 1996 Journal of Bacteriology and Virology Vol.26 No.2
Fecal samples of calf diarrhea were taken on farms in Jeju island, rotavirus was isolated and cytopathic effect (CPE) was determined after infection to MA104 cell. Morphological evaluation on electron microscopy proved it as rotavirus. Also, its infection to MA104 cell was reidentified using a fluorescence antibody method. Genotype of Jeju island bovine rotavirus (JBR) analyzed through PAGE was 4: 2: 3: 2 pattern, which was unique in bovine and that analyzed through general PAGE was somewhat different from NCDV, UK, KK3, A5-3A, 61A, B223 and similar to N stoo1-5, N culture-5 and Kawatabi (Japan). By titration after plaquing, the level was 1-3 x 10' PFU/ml, which was lower than those of NCDV and UK. Electrophoresis analysis of RNA- RNA hybridization, ELISA, and first and second PCR products of VP7 and VP4 in 1% agarose (TAE+1pl EtBr) revealed that the rotavirus was a serotype of G6P11.
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JBR(Jeju Island Bovine Rotavirus)의 Sequencing에 의한 Homology 비교에 관한 연구
이종익,谷口孝喜,김응률,류영수,박선오,송진욱,유제현,조홍찬,차광종,浦澤正三,이태협,김유성,이영건 건국대학교 동물자원연구센터 1998 動物資源硏究誌 Vol.19 No.-
본 연구는 PCR에 의해 로타바이러스의 serotype을 결정하고 염기배열의 유사성을 다른 지역에서 발견된 로타바이러스와 비교 분석하였으며 그 결과는 다음과 같다. 1. 제주도 목장에서 송아지 설사변을 채취하여 로타바이러스를 분리 후 MA104세포에 감염시켜 세포변성 효과(CPE)를 확인하였다. 2. 전자현미경에 의해 형태학적로 로타바이러스임이 판명되었으며 형광항체법에 의해 MA104세포에 감염된 것을 재확인 하였다. 3. 제주도 송아지 로타바이러스(JBR)RNA를 PAGE에 의해 genotype를 분석한 결과 bovine특유의 4:2:3:2 pattern이지만 일반 PAGE로 분석한 결과는 NCDV, UK, KK3, A5-37, 61A, B223와 차이가 있었으며, Ns-5, Nc-5, Kawatabi(Japan)와는 비슷한 경향을 보였다. 4. Plaquing 후 titer한 결과 NCDV, UK보다는 낮은 ??PFU/ml이었다. 5. RNA-RAN hybridization과 ELISA 및 VP7과 VP4의 1차, 2차 PCR 사물을 1% agarose(EtBr 1㎕의 TAE)에서 전기영동한 결과 G6P11의 serotype이었다. 6. JBR의 P serotype이 동일하게 나타난 B223과의 비교 결과 총 JVP8 750 bases 중에서 731개의 염기가 B223과 같은 것으로 나타나 염기배열의 유사성은 97.47%로 나타났고, 아미노산 배열의 유사성은 97.57%로 나타났다. This study was carried our to identify JBR's serotype by polymerase chain reaction(PCR) and to analyze homology of JBR' sequence. The results obtained in these experiments were as follows: 1. Fecal samples of calf diarrhea were taken on farms in Jeju island, rotavirus was isolated and cytopathic effect(CPE) was determined after infection to MA104 cell. 2. Morphological evaluation on electron microscopy proved it as rotavirus. Also, its infection to MA104 cell was reidentified using a fluorescence antibody method. 3. Genotype of Jeju island bovine rotavirus(JBR) analyzed thorough PAGE was 4:2:3:2 pattern, which was unique in bovine and that analyzed through general PAGE was somewhat different from NCDV, UK, KK3, A5-37, 61A, B223 and similar to Nstool-5, N cultrue-5 and Kawatabi(Japan). 4. By titration after plaquing, the level was ??PEU/ml, which was lower than those of NCDV and UK. 5. Electrophoresis analysis of RNA-RNA hybridization. ELISA, adn first and second PCR products of VP7 and VP4 in 1% agarose(TAE+1㎕ EtBr) revealed that the rotavirus was a serotype of G6P11. 6. Alignment of two(JBR and B223) amino acid sequences is 97.57% and nucleotide sequences is 97.47%.
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