• 한국식물병리학회:학술대회논문집
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• 한국식물병리학회 1999년도 정기총회 및 춘계 학술발표회
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• pp.20.2-20
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• 1999

• 원예과학기술지
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• 제18권4호
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• pp.513-517
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• 2000

우리나라 난 재배 농가에서 많이 재배하고 있는 호접란 대만 수입묘의 바이러스 감염 정도를 검정하기 위하여 플라스크묘 상태의 실생 번식 식물체를 공시하여 역전사 중합효소 연쇄반응(RT-PCR) 기술에 의해 CymMV와 ORSV의 감염 여부를 검정하였다. 호접란 식물체의 잎에서 조즙액을 추출하여 RT-PCR을 위한 total RNA로 사용하였다. $42^{\circ}C$에서 45분간 반응시켜 cDNA를 합성하였으며, $96^{\circ}C$에서 2분간 template를 예비 변성시킨 후, $96^{\circ}C$에서 30초간 template 변성, $60^{\circ}C$에서 30초간 primer 부착 및 $72^{\circ}C$에서 1분간 DNA 합성을 1cycle로 하여 총 36cycle을 반응시키고, $72^{\circ}C$에서 10분간 안정화하는 조건으로 one-step RT-PCR을 수행하였다. 바이러스 검정 결과 정도의 차이는 있으나, 40개 시료 모두 CymMV에 감염되어 있었으며, ORSV에 감염된 식물체는 없었다.

• 한국식물병리학회지
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• 제10권3호
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• pp.228-234
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• 1994

Genomic RNA was extracted from Cy strain of odontoglossum ringspot tobamovirus (ORSV-Cy) isolated from infected leaves of tobacco cv. Samsun. Size of the genomic RNA was about 6.6 kb in length. The genomic RNA was fractionated using Sephadex G-50 column chromatography into 2 fractions. They were polyadenylated at their 3'-end using E. coli poly(A) polymerase. Polyadenylated viral RNA was recovered by oligo (dT) primer adapter containing NotI restriction site and Moloney murine leukemia virus SuperScript reverse transcriptase (RNase H-). Second-strand cDNA was synthesized by using E. coli DNA ligase, E. coli DNA polymerase I and E. coli RNase H. Recombinant plasmids containing cDNAs for ORSV-Cy RNA ranged from about 800 bp to 3,000 bp. Among the selected 238 recombinants, pORCY-124 clone was the largest one covering 3'-terminal half of the viral RNA. This clone contained two restriction sites for EcoRI and XbaI and one site for AccI, AvaI, BglII, BstXI, HindIII, PstI, and TthIII 1. respectively. The clone contained partial viral replicase, a full-length movement protein and a complete coat protein genes followed by a 3' untranslated region of 414 nucleotides based on restriction mapping and nucleotide sequencing analyses. Clones pORCY-028, -068, -072, -187 and -224 were overlapped with the pORCY-124. Clones pORCY-014 and -095 covered 5' half upstream from the middle region of the viral RNA, which was estimated based on restriction mapping and partial sequence analysis. Constructed cDNA library covered more than 90% of the viral genome.