Applied Biological Chemistry
- Volume 38 Issue 1
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- Pages.55-62
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- 1995
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- 2468-0834(pISSN)
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- 2468-0842(eISSN)
Identification of a Potexvirus in Korean Garlic Plants
한국 마늘 Potexvirus의 cDNA 유전자 분리 및 분포에 관한 연구
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Song, Jong-Tae
(Department of Agricultural Chemistry, Seoul National University) ;
- Choi, Jin-Nam (Department of Agricultural Chemistry, Seoul National University) ;
- Song, Sang-Ik (Department of Agricultural Chemistry, Seoul National University) ;
- Lee, Jong-Seob (Department of Molecular Biology, Seoul National University) ;
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Choi, Yang-Do
(Department of Agricultural Chemistry, Seoul National University)
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송종태
(서울대학교 농업생명과학대학 농화학과) ;
- 최진남 (서울대학교 농업생명과학대학 농화학과) ;
- 송상익 (서울대학교 농업생명과학대학 농화학과) ;
- 이종섭 (서울대학교 자연대학 분자생물학과) ;
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최양도
(서울대학교 농업생명과학대학 농화학과)
- Published : 1995.02.28
Abstract
To understand the molecular structure of Korean garlic viruses, cDNA cloning of virus genomic RNA was attempted. Virus particles were isolated from virus-infected garlic leaves and a cDNA library was constructed from garlic virus RNA. One of these clones, S81, selected by random sequencing has been identified as a member of potexvirus group other than potyvirus and carlavirus. The clone is 873 bp long contains most of the coat protein (CP) coding region and 3'-noncoding region including poly(A) tail. A putative polyadenylation signal sequence (AAUAAA) and the hexanucleotide motif (ACUUAA), a replicational cis-acting element conserved in the 3'-noncoding region of potexvirus RNAs are noticed. The clone S81 shows about 30-40% identity in both nucleotide and amino acid sequences with CPs of potexviruses. The genome size of the virus was analysed to be 7.46 knt by Northern blot analysis, which was longer than those of other potexviruses. The open reading frame encoding CP was expressed as a fusion protein (S81CP) in Escherichia coli and the recombinant protein was purified by immobilized metal binding affinity chromatography. Polyclonal antibody was raised against S81CP in rabbit to examine the occurrence of garlic potexvirus in Korean garlic plants by immunoblot analysis. Two virus protein bands of Mr 27,000 and 29,000 from garlic leaf extract of various cultivars reacted with the antibody. It was shown that Mr 27,000 band might not be a degradation product of Mr 29,000 band, suggesting that two types of potexvirus different in size of coat protein could exist in Korean garlic plants.
한국 마늘 바이러스의 유전자 구조와 병 발생 메카니즘을 연구하기 위하여, 바이러스가 감염된 마늘잎으로부터 바이러스 입자를 분리하고 RNA를 추출하였다. 그 virus RNA를 이용하여 마늘 바이러스 cDNA 유전자 은행을 만들어 일부 clone의 염기 서열을 결정하였다. 여기에서 얻은 cDNA clones 중에서 poly(A) tail을 갖는 clone S81를 분리하고 873 bp의 전체 염기서열을 결정하였다. Clone S81의 염기서열을 다른 식물 바이러스와 비교한 결과 potexvirus의 껍질단백질 부분의 염기서열과