• 식물병연구
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• 제23권1호
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• pp.65-68
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• 2017

2단계의 nested PCR 방법을 이용하여 보리 및 벼 재배 토양에서 Barley yellow mosaic virus (BaYMV)를 검출하였다. BaYMV 분절 RNA1 외피단백질 영역의 특이 프라이머로 1차 PCR을 하고 내부서열로부터 작성된 프라이머로 2차 PCR을 실시하여 확보된 372 bp의 PCR 산물이 BaYMV 외피단백질 영역과 98%-100% 염기서열이 일치하여 BaYMV를 검출할 수 있음을 확인하였다. 이 결과는 토양으로부터 BaYMV 검출에 관한 최초의 보고이며 토양전염성 바이러스의 정확한 진단과 예찰에 적용될 수 있을 것으로 생각한다.

• 식물병연구
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• 제24권2호
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• pp.170-173
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• 2018

춘천과 충주에서 퇴록현상과 모자이크 병징이 뚜렷한 닭의 장풀을 채집하여 바이러스 분리 동정하였다. 해당 바이러스는 RT-PCR과 RFLP 분석결과 CMV로 동정되었으며, 닭의장풀 CMV 분리주들은 서로 다른 서브그룹으로 존재하고 있는 것이 확인되었다. 외피단백질유전자의 염기서열을 분석한 결과, 각 CMV 분리주들은 CMV-Fny와 비교 하였을 때 CMV-Co와 96.3%, CMV-Co2는 96.3%, CMV-Co3는 95.9%의 유사성을 나타내었다. 또한 기주 반응 분석 결과 닭의장풀 분리주들은 대조바이러스(CMV-Fny)와 달리 C. pepo에서 전신 괴사병징이 특징적으로 나타났으며 Physalis angulata과 N.rustica의 경우 닭의장풀 분리주 간에 병징의 차이를 보였다. 이들 결과를 통해, 국내 닭의장풀에서 분리된 CMV들에서 유전적, 생물학적 다양성이 있는 것으로 확인되었다.

• The Plant Pathology Journal
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• 제18권3호
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• pp.130-137
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• 2002

Potato virus Y (PVY) is one of the most important viruses in many field crops in Korea. In this study, 31 PVY isolates were isolated from infected potato (Solanum tuberosum), tobacco (Nicotiana tabacum), pea (Pisum sativum), and weeds (Veronica persica, Lamium amplexicause and Capsella bursa-pastoris) showing different mosaic symptoms in Jeonbuk, Chungnam, Gangwon, and Gyeongbuk areas in Korea. The 640 nucleotide region containing the C-terminal portion of coat protein (CP) gene and 3'non-translated region (NTR) was amplified by reverse transcription-polymerase chain reaction (RT-PCR) using PVY-specific oligonucleotide primers. Sequence analyses of the amplified DNA fragments showed that the C-terminal portion of CP gene was not significantly different from that of previously reported PVY strains from potato (PVY-OK and -T) and tobacco (PVY-VN) in Korea. Homologies of the deduced CP amino acid sequences were 93.3-99.0% to corresponding regions of the other PVY strains including PV $Y^{N}$, PV $Y^{o}$ , PV $Y^{OK}$ , PV $Y^{T}$ , and PV $Y^{VN}$ . In contrast the sequences located at the 3'-NTR showed more diverse sequence homologies (76.4-99.7%). These results indicate that the C-terminal portion of the CP gene was relatively conserved while sequences at the 3'NTR were more diverse and variable over the host species and the regions where they were isolated.e isolated.

• 한국어병학회지
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• 제25권2호
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• pp.111-116
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• 2012

본 연구에서는 2006-2008년 남해안 일대의 해상가두리에서 사육중인 능성어에서 발생하는 viral nervous necrosis (VNN)의 발생양상을 조사하였다. VNN은 사육 수온이 $24-26^{\circ}C$ 범위인 8월부터 발생하기 시작하여 수온이 $20-25^{\circ}C$ 범위인 9-10월까지 지속되었고, 성어보다는 치어에서 폐사율이 높게 나타나는 것으로 확인되었다. 폐사되는 패턴으로는 급성으로 인한 대량폐사와 소량으로 지속적으로 폐사되는 경우가 확인되었다. 능성어로부터 분리된 NNV 분리주들의 coat protein gene을 계통분석한 결과, 분리주들은 모두 RGNNV 유전자형에 속하였다. 이상의 결과로 능성어 양식장에서의 VNN은 RGNNV type의 NNV에 의해 여름철 7-9월(사육수온: 약 $24^{\circ}C$)에 치어뿐만 아니라 성어에서 발생하는 것으로 확인되었다.

• The Plant Pathology Journal
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• 제16권6호
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• pp.306-311
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• 2000

A new strain of Cucumber mosaic virus (CMV) from easter lily (Lilium longiflorum), Ly2-CMV, was identified and compared to the well-characterized Mf-CMV (subgroupⅠ) and LS-CMV (subgroupⅡ) by host reaction in several indicator plants, dsRNA analysis, serological property, RT-PCR analysis, restriction enzyme profile of the PCR products and nucleotide sequence of coat protein (CP) gene. Remarkable differences in symptoms of Ly2-CMV were found between Mf-CMV or LS-CMV in tobacco plants and Datura stramoinium. Ly2-CMV induced small necrotic ringspots on the inoculated leaves of Nicotiana tabacum cvs. Xanthi nc and Burley 21 and D. stramonium, and failed to infect these species systemically. Of the indicator plants tested, N. benthamiana only reacted with systemic infection by inoculation of Lr2-CMV. In experiments of dsRNA analysis, serology and RT-PCR of CP gene, Ly2-CMV was come within subgroupⅠ CMV. However, restriction enzyme analysis of the PCR products using MspⅠ showed that Ly2-CMV was distinct to Mf-CMV. The CP gene of Ly2-CMV contains 657 nucleotides, and the nucleotide sequence is similar to that of Mf-CMV. There is also a high degree of conservation between their putative gene products in Ly2-CMV and Mf-CMV, with five amino acid changes in the 218 amino acids of the CPs.

• 식물조직배양학회지
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• 제24권5호
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• pp.313-317
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• 1997

ND860-2, Norchip, Russet Norkotah, Goldrush 및 Norgueen Russet 등 5개 품종의 감자 잎 조직을 감자 바이러스 Y(PVY)의 외피단백질 (CP) 유전자를 운반하는 Agrobacterium tumefaciens을 이용하여 형질전환시켰다. ND860-2와 Norchip품종과 같은 흰색 감자에서 재분화율과 형질전환율이 높게 나타났다. 형질전환체를 선발한 후 CP의 생성을 western blot으로 확인한 결과 30 kD의 밴드가 나타났다. 온실에서 재배된 형질전환체는 한 개체를 제외하고는 표현형에 있어서는 기존의 품종과 다르지 않았다. 인위적으로 감자 바이러스 Y를 감염시킨 15일 후의 검사결과 control구는 80%의 감염율을 보인 반면 형질전환체들은 38-58%의 감염율을 보였다. 30일 후의 검사 결과는 모든 5품종의 control구에서 100%의 감염을 보였으나 형질전환체는 7개체가 저항성을 보였다. 마지막 85일 후의 검사 결과 4개의 저항개체를 발견하였으며 이 개체들은 현재 계속 연구가 진행 중에 있다.

• The Plant Pathology Journal
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• 제15권5호
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• pp.270-279
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• 1999

cDNAs complementary to the 3'-terminal regions of two potyvirus genomes were cloned and sequenced. The clone G7 contains one open reading frame (ORF) of 1,338 nucleotides and a 3' untranslated region (3'-UTR) of 403 nucleotides at the 3'-end excluding the 3'end poly(A) tail. The putative viral coat protein (CP) shows 55%-92% amino acid sequence homology to those of Allium potyviruses. The genome size of the virus was analyzed to be about 9.0 kb by Northern blot analysis. Five cDNA clones were screened out using GPV2 oligonucleotide as a probe. One of these clones, DEA72, which has a longest cDNA insert, contains one ORF of 1,459 nucleotides and a 3'-UTR of 590 nucleotides at the 3'-end excluding the 3'-end poly(A) tail. The putative viral CP shows 57%-88% amino acid sequence homologies to those of Allium potyviruses. The genome size of the virus was analyzed to be about 9.6 kb by Northern blot analysis. The results of immunoblot and Northern blot analyses suggest that almost all of the tested garlic plants showing mosaic or streak symptoms are infected with DEA72-potyvirus in variable degrees but rarely infected with G7-potyvirus in variable degrees but rarely infected with DEA72-potyvirus in variable degrees but rarely infected with G7-potyvirus. Immunoelectron microscopy using anti-DEA72 CP antibody shows that this potyvirus is about 750 nm long and flexuous rod shaped.

• 한국식물병리학회:학술대회논문집
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• 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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• pp.141.1-141
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• 2003

Forty six isolates of bean common mosaic virus (BCMV) collected from azuki bean, mungbean, kidney bean, cowpea, broad bean and peanut were classified into three groups based on biological, serological, cytopathological, and molecular characteristics. Group I induced vein-banding symptoms in cowpea which was similar to those produced by the BCMV-cowpea strain. Group II caused mosaic symptoms in azuki bean but not in peanut and tobacco. Since this character was different from that of previously described BCMV strain, group II may not belong to BCMV GroupIII induced vein-clearing symptoms in azuki bean, kidney bean and peanut, which are typical symptoms for BCMV-peanut stripe virus strain. Virus inclusion patterns of BCMV groups were similar to those of Potyvirus subdivision III with the scroll, pinwheel and long laminated inclusions. However, the inclusions of laminated aggregates were never observed in mungbean isolates. Multiple alignment as well as cluster dendrograms of 3'noncoding region (3'-NCR) and a part of coat protein gene (CP) suggested that group I belongs to the BCMV-cowpea strain, group II to the BCMV-azuki bean strain, and group III to the BCMV-peanut stripe virus strain. Since molecular phylogenesis of BCMV based on nucleotides of 3'-NCR and coat protein differed from the grouping based on virulence differentiation, and BCMV groups are more closely related to each other with the same host origin, other characteristics of those strains are under investigation.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2001년도 추계학술발표대회
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• pp.829-832
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• 2001

Phage antibody 생산방법은 유전자 조작에 의하여 phage coat에 항체를 발현시키는 방법이며 그 library를 이용하여 일반 hybridoma 방법에 의해 항체를 생산하기 어려운 성분 (예: 독성물질, 면역화가 잘 안되는 물질 등) 에 대해 적용할 수 있는 장점을 제공한다. 본 연구진은 Griffin.1 antibody library의 positive control을 통해 phage 표면에 항체가 발현되는 지의 여부를 ELISA test를 통하여 확인하였다. 또한 human serum albumin을 모델분석물질로 사용하여 $1{\sim}4$차까지의 bio-panning 을 실시하였고 각 단계에서 증폭된 phage를 가지고 ELISA test를 한 결과 신호가 점진적으로 증가하는 data를 얻을 수 있었다. 이것을 통해 phage library로부터 특정 항원에 대한 monoclonal antibody를 생산할 수 있다는 사실을 검증하였다.

• Applied Microscopy
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• 제1권1호
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• pp.11-17
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• 1969

Conidio spores of Aspergillus niger (strain No. NRRL 330) cultured on potato dextrose agar media were studied by electron microscopy, using the thin sectioning techniques. Conidio spores to be sectioned were fixed by triple methods with $K_2Cr_2O_7$, Glutaraldehyde and $OsO_4$. After dehydrated with alcohol, the specimens were embedded in metacrylate and epon resin media, and thinly sectioned by Porter-Blum MT-2. After sectioned these specimens were negative-stained with uranyl acetate and observed. by Hitachi HS-6 electron microscope. The results of this experiment were summarized as follows. 1. The structures of spore ,wall system seem to be formed 4 layers; exosporium, basal layer, spore coat and unit cell membrane. The protuberance of spore surface that was looked like hair appears to be protrusived from the basal layer. 2. The 3 layers of unit cell membrane was constituted outer layer membrane, inner layer membrane and inter-mediate light layer. 3. The structures of intra cytoplasmic membrane appear as spiral form which was consisted of 3 layers membrane system; outer membrane, inner membrane, and intermediate layer, which has pits. 4. The cement substance of spore coat and cortex may be changed quantitatively by physiological state in cell. 5. In some cases, we observed that the ribosome was transformed into poly ribosome group, and the storage materials and the protein crystals were changed variously. It. has been suggested that the morphological change of some cytoplasmic materials may be caused by some specialized function of the physiological stage.