• Korean Journal of Agricultural Science
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• v.43 no.1
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• pp.21-27
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• 2016

Apple stem grooving virus (ASGV), Apple chlorotic leaf spot virus (ACLSV), and Apple stem pitting virus (ASPV) have been known to induce top working disease causing economical damage in apple. Occurrences of these three viruses in pome fruit trees, including apple, have been reported around the world. The transmission of the three viruses was reported by grafting, and there was no report of transmission through mechanical contact, insect vector, or seed except some herbaceous hosts of ASGV. As RNA extraction methods for fruit trees, Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR) and multiplex RT-PCR techniques have been improved for reliability and stability, and low titer viruses that could not be detected in the past have become detectable. We studied the seed transmission ability of three apple viruses through apple seedling diagnosis using RT-PCR. Nineteen seeds obtained from commercially grown apple were germinated and two of the resulting plants were ASGV positive. Seven clones of the amplified ASGV coat protein (CP) genes of these isolates were sequenced. Overall sequence identities were 99.84% (nucleotide) and 99.76% (amino acid). Presence of a previously unreported single nucleotide and amino acid variation conserved in all of these clones suggests a possible association with seed transmission of these 'S' isolates. A phylogenetic tree constructed using ASGV CP nucleotide sequences showed that isolate S sequences were grouped with Korean, Chinese, Indian isolates from apple and Indian isolates from kiwi.

• Journal of Life Science
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• v.12 no.4
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• pp.496-503
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• 2002

To develop a yeast strain of Pichia pastoris producing an antibacterial peptide, we have attempted the expression and secretion of an insect defensin. The nucleotide sequences corresponding to mature defensin were chemically synthesized by 6 oligomers, assembled in vitro and the synthesized gene was identified by nucleotide sequencing. The prepro sequence of yeast mating factor $\alpha$1 and the defensin gene were recombined into a Pichia expression vector, pPIC9K. The resulting plasmid, pPIDE, was transformed into P. pastoris GSl15 and transformants selected on histidine-deficient minimal plates were tested for antibacterial activity against Micrococcus luteus. Four strains with different antibacterial activity were selected for further analysis. Southern hybridization and RT-PCR verified the defensin gene was maintained and transcribed in a host. Four strains were cultivated in YPD broth for 96 hours to compare cell growth and antibacterial activity, They showed no difference in cell growth, however, each strain showed different antibacterial activity pattern with culture time. The maximal activity was about 550 AU/ $m\ell$.

• Environmental Analysis Health and Toxicology
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• v.15 no.4
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• pp.139-145
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• 2000

The Flavin-containing monooxygenase (FMOs) (EC1.14.13.8) are NADPH-dependent flavoenzymes that catalyze oxidation of soft nucleophilic heteroatom centers in a range of structurally diverse compounds, including foods, drugs, pesticides, and other xenobiotics. In humans, FMO3 is quantitatively a major human liver monooxygenase. In the present study, the baculovirus expression vector system was used to overexpress human FMO3 in insect cells for catalytic studies. Six commercially available organic selenium compounds were examined for substrate activity with microsomes isolated from Spodoptera frugiperda (Sf)9 cells infected with human FMO3 recombinant baculovirus. While none of the aromatic heterocyclic selenides tested showed detectable activity, all dialkyl- and alkylaryl-selenides free from ionic groups catalyzed the NADPH- and O$_2$-dependent oxidation. Kinetic constants demonstrate that (based on Km) dialkyl-and alkylaryl- selenides are better substrates for human FMO3 than analogous nitrogen or sulfur compounds .

• Microbiology and Biotechnology Letters
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• v.25 no.4
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• pp.359-366
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• 1997

The usefulness of host range expanded recombinant viruses for economical viral insecticide and expression vector system has been studied. Host range expanded recombinant viruses, RecS-B6 and RecB-8, constructed by cotransfection of Bombyx mori nuclear polyhedrosis virus (BmNPV) and Autographa californica NPV (AcNPV), and a host range expanded AcNPV recombinant, Ac-BH, constructed by substitution of the 0.6Kb fragment of the BmNPV helicase gene were compared. The restriction enzyme digestion patterns showed that RecS-B6 and RecB-8 had expanded host ranges by genomic recombination and were more similar to genome of AcNPV than that of BmNPV. SDS-PAGE and PCR analysis showed that the polyhedrin gene of RecS-B6 and RecB-8 was derived from BmNPV genomic DNA. The morphology of polyhedra of recombinant viruses showed a slight difference between the two host cells, Sf and BmN cells, indicating that the morphology of polyhedra was influenced by host cells. The bioassay data for insect larvae showed that Ac-BH, compared to wild type viruses, had superior pathogenicity against Bombyx mori larvae but inferior pathogenicity against Spodoptera exigua larvae. Although the pathogenicity was lower than that of wild type viruses in both larvae, RecS-B6 showed the pathogenicity in both larvae. These results suggested that Ac-BH was a less useful economical insecticide than random genomic recombinant virus RecS-B6.

• Journal of Microbiology and Biotechnology
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• v.19 no.2
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• pp.194-203
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• 2009

$HpaG_{Xooc}$, from rice pathogenic bacterium Xanthomonas oryzae pv. oryzicola, is a member of the harpin group of proteins, eliciting hypersensitive cell death in non-host plants, inducing disease and insect resistance in plants, and enhancing plant growth. To express and secret the $HpaG_{Xooc}$ protein in Bacillus subtilis, we constructed a recombinant expression vector pM43HF with stronger promoter P43 and signal peptide element nprB. The SDS-PAGE and Western blot analysis demonstrated the expression of the protein $HpaG_{Xooc}$ in B. subtilis. The ELISA analysis determined the optimum condition for $HpaG_{Xooc}$ expression in B. subtilis WBHF. The biological function analysis indicated that the protein $HpaG_{Xooc}$ from B. subtilis WBHF elicits hypersensitive response(HR) and enhances the growth of tobacco. The results of RT-PCR analysis revealed that $HpaG_{Xooc}$ induces expression of the pathogenesis-related genes PR-1a and PR-1b in plant defense response.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1995.10a
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• pp.107-113
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• 1995

Basement membrane laminin is a multidomain glycoprotein that interacts with itself, heparin and cells. The distal long arm plays major cell and heparin interactive roles. The long arm consists of three subunits (A, B1, B2) joined in a coiled-coil rod attached to a terminal A chain globule (G). The globule is in turn subdivided into five subdomains (Gl-5). In order to analyze the functions of this region, recombinant G domains (rG, rAiG, rG5, rGΔ2980-3028) were expressed in Sf9 insect cells using a baculovirus expression vector. A hybrid molecule (B-rAiG), consisting of recombinant A chain(rAiG) and the authentic B chains (E8-B)was assembled in vitro. The intercalation of rAiG into E8-B chains suppressed a heparin binding activity identified in subdomain Gl-2. By the peptide napping and ligand blotting, the relative affinity of each subeomain to heparin was assigned as Gl> G2= G4> G5> G3, such that G1 bound strongly and G3 not at all. The active heparin binding site of G domain in intact laminin appears to be located in G4 and proximal G5. Cell binding was examined using fibrosarcoma Cells. Cells adhered to E8, B-rAiG, rAiG and rG, did not bind on denatured substrates, poorly bound to the mixture of E8-B and rG. Anti-${\alpha}$6 and anti-${\beta}$1 integrin subunit separately blocked cell adhesion on E8 and B-rAiG, but not on rAiG. Heparin inhibited cell adhesion on rAiG, partially on B-rAiG, and not on E8. In conclusion, 1) There are active and cryptic cell and heparin binding activities in G domain. 2) Triple-helix assembly inactivates cell and heparin binding activities and restores u6131 dependent cell binding activities.

• International Journal of Industrial Entomology and Biomaterials
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• v.7 no.2
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• pp.127-131
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• 2003

Protein disulfide isomerase (PDI) found in the endoplasmic reticulum (ER) catalyzes disulfide bond exchange and assists in protein folding of newly synthesized proteins. PDI also functions as a molecular chaperone and has been found to be associated with proteins in the ER. In addition, PDI functions as a subunit of two more complex enzyme systems: the prolyl-4-hydroxylase and the triacylglycerol transfer proteins. A cDNA that encodes protein disulfide isomerase was previously isolated from Bombyx mori (bPDI), in which open reading frame of 494 amino acids contained two PDI-typical thioredoxin active site of WCGHCK and an ER retention signal of the KDEL motif at its C-terminal, and we report its functional characterization here. This putative bPDI cDNA is expressed in insect Sf9 cells as a recombinant proteins using baculovirus expression vector system. The bPDI recombinant proteins are successfully recognized by antirat PDI antibody, and shown to be biologically active in vitro by mediating the oxidative refolding of reduced and scrambled RNase. This suggests that bPDI may play an important role in protein folding mechanism of insects.

• Journal of Sericultural and Entomological Science
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• v.13 no.1
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• pp.17-22
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• 1971

A series of experiments were conducted from 1968 to 1970 to find the transmission effects of the dwarf disease of mulberry tree in connection with inlay grafting and insect vector with the following conclusions: 1. The inlay-grafting of sound scions on diseased roots showed a 30 per cent transmission rate among the successful grafts. The inlay-grafting of diseased scions on sound roots showed a 14 per cent transmission rate among the successful grafts ir the case of scions cut and stored before wintering but no transmission effect in the case of scions prepared after wintering, There were some stocks, among the unsuccesful grasfts of diseased scions upon sound roots, that sprouted out new buds, no transmission effects were found in them, 2. Leafhoppers (Hishimonus disciguttus Walker) collected from areas with no dwarf disease infection were grouped into three plots and let to feed on mulberry tree severely infected by dwarf disease for 7 days, 14 days and 21 days respectively. After these preliminary treatment, the leafhoppers were recollected to be fed on the sound seedling divided into three plots for 10 days, 15 days and 20 days respectively to find the transmission effects. The results are that the 7 day plot showed no transmission effect whereas the 14 day plot showed a 22 per cent disease rate and the 21 day plot 61 per cent rate indicating that the longer the feeding period is the higher the transmission rate.

• Korean journal of applied entomology
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• v.29 no.1
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• pp.14-19
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• 1990

Nematodes and insects associated with dead trees were surveyed on the 238 dead trees belonging to 19 tree species in Pusan, Gyeongnam, Gyeongbuk, Chunnam, and Chonbuk provinces from April to September of 1989. Pine wood nematode, Bursaphelenchus xylophilus was found only in Pusan but B. mucronatus was collected at Chinju and Chinhae. The 13 nematode species in 9 genera were identified. Out of them, Diplogasteroides dimidius, Rhabdontolaimus adephagus, R. janae, Mikoletzkya diluta, M. ruminis, M. langcaudaa, Parasitorhabditis hylurgi, Panagrolaimus concolor, Panagrodontus dentatus, Prothalonema intermedium, and marcrolaimus canadensis were recorded for the first time in Korea. Insects collected from dead trees were 5 orders, 9 families, 25 genera, and 27 species. of them, the Coleoptera were the most collected insects by the 3 families, 19 genera and 22 species. The Scolytidae were 12 species in 10 genera. Hypothenemus eruditus was firstly collected from Campylotropis macrocarpa, Lespedeza maximowizi, Forsythia ovata, Meliosma oldhami, Securinega suffruticosa, Broussonetia kazinoki, and Cornus walteri. The maximum number of pine wood nematode was separated from the abdomen of Monchamus alternatus, the pine woodnematode vector. The maximum number of nematodes per an adult of M. alternatus was 127,535, minimum 2,616, and average 42,817.

• Journal of Plant Biology
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• v.3 no.1
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• pp.32-38
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• 1960

Since the peculiar virus disease of chinese date tree (Zizyphus jujuba Mill. var. inermis Rehd.) has been noted in South Korea around 1950, 70% to 80% of the economically important trees have been either completely destroyed or infected with the virus, severe damage has been noted, particularly, across the area ranged from middle east to the middle part of Korea, including Seoul area. Yoon-Koock-Byung in 1958 first reported the disease and descirbed it might be caused by a kinds of yellows. But he did not conform in his paper that the disease is pecisely caused by yellows virus. The authors, hereby intend to identify the true cause of the desease of the chinese data tree by studying the external symptoms of the disease and the internal morphological characteristics of the diseaset plant which shows various abnormalities in contrast to the healthy checks. In view of fact that leaves of the infected plants become yellowish in color similar to the peach yellows, aster yellows, it is likely to be identifiable as the common yellows. Furthermore, the abnormal characteristics observed by the authors are as follow: The floral organs such as petals, sepals, stamens, and pistil turn into vegetative leaves, the leaves on heavily infected plant appear as small sized one and also showing as a common witch's broom like symptom. There are also an occuring of numerous advantitious shoots developed from both of stems and roots. The amount of photosynthetic starch grains increases in parenchymatous cells, necrosis takes place in mesophyll, Particularly, Palisade Parenchyma in the leaves of infected plants are distinguished in contrast to the healthy checks. From the symptoms and the present experimetns described above, the authors are believed that the disease of chinese data tree is not caused by the yellows. It appears the disease is rather similar to the symptoms of sandal spike virus which was noted in India early in this centry. But the host plant of standal disease, Santalum albun L. and the insect vector, Jassus indicus Wal., have never been reported in Korean flora and the founa. The termperature and the otehr environmental factors is quite different Korea and India. Thus the authors believe that the peculiar disease must be an endemic new virus origin in Korea and must be called as "shoot cluster disease of chinese date tree."