• Applied Biological Chemistry
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• v.38 no.1
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• pp.55-62
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• 1995

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.

• Journal of Pharmaceutical Investigation
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• v.31 no.3
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• pp.185-190
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• 2001

Human papillomavirus (HPV) infection is known to cause cervical cancers. Human papillomavirus-like particles (VLP) have been studied as preventive vaccines of cervical cancers. To develop VLP as a therapeutic gene carrier, we studied the method to encapsulate cytokine genes in virus-like particles. HPV type 16 capsid L1 genes were amplified by polymerase chain reaction and cloned into T vector. L1 gene was then inserted into baculovirus transfer vector. The clone of baculovirus encoding L1 gene was isolated and used to express L1 protein in Sf 21 insect cells. VLP were purified by CsCl density gradient and ultracentrifugation. VLP were disassembled to capsomer units by treatment of a reducing agent. Given that interleukin-2 (IL-2) genes have been used in anticancer gene therapy and as a molecular adjuvant, IL-2 cytokine plasmids were chosen as a model gene. IL-2 plasmids were incubated with the disassembled capsomer suspension. To reassemble the particles, the mixture of capsomers and cytokine plasmids was dialyzed. The disassembly and reassembly of VLP were confirmed by transmission electron microscopy. The entrapment of cytokine plasmids in reassembled VLP was tested by the stability of plasmids against DNase I. After treatment of reassembled virus-like particles with DNase I, discrete IL-2 DNA band was observed. Our results indicate that IL-2 cytokine plasmid (3.5 kb size) can be encapsulated in the virus-like particles, suggesting the potential of VLP as a gene delivery system. Moreover, VLP containing the adjuvant cytokine plasmids might function as more effective subunit vaccines.

• Research in Plant Disease
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• v.20 no.4
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• pp.307-313
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• 2014

In May 2013, an angel's trumpet leaves showing mosaic and malformation symptoms were collected from Suwon city, Gyeonggi-do. An analysis of the collected sample by transmission electron microscopy observation showed filamentous rod particles of 720-800 nm in length. On the basis of the these observations, we performed PCR against three reported Potyviruses (Brugmansia mosaic virus, Colombian datura virus and Brugmansia suaveolens mottle virus), and the sample was positive for BruMV. Pathogenicity and host range test of BruMV was determined by mechanical inoculation. Solanaceae (tobacco, tomato and eggplant) and Amaranthaceae (ground cherry) appeared typical virus symptoms. To determine coat protein of this virus, we designed specific primer pairs, and performed PCR amplification, cloning, and sequencing. Phylogenetic analysis showed that BruMV-SW was most closely related to BruMV isolate SK. Comparison of the BruMV-SW coat protein nucleotide sequences showed 92% to 99% identities to the other BruMV isolates.

• The Korean Journal of Pesticide Science
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• v.5 no.2
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• pp.45-50
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• 2001

In recent, non virus-induced mosaic symptoms(NVMS) on potato leaves were observed in the seed potato fields, and its incidence rate was $5{\sim}20%$ nationwide. It made difficult to rogue out virus-infected plants, and caused much arguments between seed potato production farmers and seed potato inspectors. The objectives of these experiments were to find out the causes of NVMS, and also to induce mosaic symptom(phytotoxicity) on potato plants by treatment of several herbicides. No significant correlations were found between incidence rates of NVMS and values from soil analyses; soil pH, soil EC, organic matter content, and contents of inorganic constituents($P_2O_5,\;NO_3$, Ca, Mg, K) in the soil around the potato planted. The examinations by ELISA, virus indicator plants, and TEM showed that NVMS on potato leaves was not caused by the viruses infection. But, the use of herbicides could induced the NVMS on potato leaves. The incidence rates of potato treated with pendimethalin linuron of 400 mL/10 a, pendimethalin of 200 mL/10 a, pendimethalin.oxadiazon of 300 mL/10 a, and control were 61.1%, 47.2%, 19.4%, and 1.4%, respectively. Based on these results, we confirmed that the treatment of pendimethalin alone and in mixture with other herbicides were the reason of NVMS on potato leaves. The yields among test plots were similar except dicamba treated plot, which decreased by about 23% compared to control plot. When their progenies harvested in 1999 were planted in the following season, no symptoms of mosaic were observed.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.32 no.1
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• pp.62-67
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• 1999

Infection rates of oyster ovarian parasite, Marteiliodes chungmuensis and productivity of the oyster shellstock infected with the parasite were investigated at the main seed collection areas in the southern coast of Korea where the extreme bad seed collection of oyster occurred in 1992 and 1993 to evaluate the cause of the bad seed collection. Additionally, the bacterial flora of the sea water and oyster lana were examined to identify the shellfish larva pathogenic bacteria like Vibrio sp. and Pseueomonas sp. In August 1992 to September 1993, infection rate of oyster ovarian parasite, M. chungmuensis at Tongyong, Kyongsangnam province, and Yosu, Chollanam province where the bad seed collection occurred, were $11.8\~100\%$ and $14.3\~100\%$, respectively. But the parasite was not detected in the shellstock collected at Daechon, Chungchongnam province. While a virus-like particle was identified in the cytoplasm of the egg infected by the parasite. The parasite infected egg was not able to fertilize completely. Uninfected egg in the gonad contaminated by the parasite could be able to fertilize but showed an abnormal development till D-shaped larva and then, died of necrosis after D-shaped lana. And some lana developed from low lipid content egg could not develop to the spat and died after the early umbo stage. The predominant bacteria in the oyster lana collected at bad seed collection areas were Pseudomonas sp. and Pseudomonas like bacteria and the occupancy rates were $53.3\~87.1\%$.