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EHP와 RRV의 Coinfection에 의한 Reassortants 선발에 관한 연구
유제현,조홍찬,송진욱,김유성,이영건,박선오,이종익,김응률,차광종,류영수,Harry B. Greenberg 건국대학교 동물자원연구센터 1998 動物資源硏究誌 Vol.19 No.-
본 연구는 두 종류의 rotavirus strain MA104 cell 상에서 동시감염시킨 후 gene이 서로 교차된 새로운 reassortants를 선발하기 위해 수행하였으며, 수행한 결과는 다음과 같다. 1. EHP T 와 RRV를 MA104 세포에 감염시켜 cytopathic effect(CPE)를 확인하였으며 확인된 virus를 수확하였다. 2. Coinfection의 mobility of infection은 각각 EHP T는 0.3(3*10?? PFU), RRV는 2.5(2.5*10?? PFU)이었다. 3. EHP T와 RRV를 coinfection하여 plaque assay에 의해 형성된 plaque들을 pick up하였다. 4. EHP T와 RRV를 동시감염시켜 얻어진 plaques를 MA104 cell에 재감염한 후 grow up하여 수확하였다. 5. Plaques를 24 Well의 MA104 cell에 감염하여 수확한 후 dsRNA를 추출하여 전기영동에 의해 26개 reassortants를 선발하였다. This study was carried out to select new reassortants whose genes were exchanged by confection of two strains. The results obtained in these experiments were as follows: 1. Cytopathic effect was checked out in the experiment in which made MA104 cells infected by EHP T (murine rotavirus) and RRV(simian rhesus rotavirus). And then the viruses were obtained from the procedure. 2. Mobility of infection(MOI) of EHP T and RRV were 0.3(3*10?? plaque forming units) and 2.5(2.5*10?? plaque forming units)respectively. 3. After coinfection of EHP T and RRV, plaques which formed in plaque assay were picked up. 4. Coinfected EHP T and RRV plaques were grown up in 24 well of MA104 cell and then grown-ups or those were obtained. 5. By polyacrylamide gel electrophoresis(PAGE) of dsRNA extracted above, 26 reassortants were selected.