• The Journal of Korean Society of Virology
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• v.27 no.2
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• pp.239-255
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• 1997

Expression of the cDNA of the VP1 gene on the genome RNA B segment of infectious pancreatic necrosis virus (IPNV) DRT strain in E. coli and a recombinant baculovirus were carried out. The VP1 gene in the pMal-pol clone (Lee et al. 1995) was cleaved with XbaI and transferred into baculovirus transfer vector, pBacPAK9 and it was named pBacVP1 clone. The VP1 gene in the pBacVP1 clone was double-digested with SacI and PstI and then inserted just behind T5 phage promoter and the $6{\times}His$ region of the pQE-3D expression vector, and it was called pQEVPl. Again, the $6{\times}$His-tagged VP1 DNA fragment in the pQEVP1 was cleaved with EcoRI and transferred into the VP1 site of the pBacVP1, resulting pBacHis-VP1 recombinant. The pBacHis-VP1 DNA was cotransfected with LacZ-Hyphantria cunea nuclear polyhedrosis virus (LacZ-HcNPV) DNA digested with Bsu361 onto S. frugiperda cells to make a recombinant virus. One VP1-gene inserted recombinant virus was selected by plaque assay. The recombinant virus was named VP1-HcNPV-1. The $6{\times}$His-tagged VP1 protein produced by the pQEVP1 was purified with Ni-NTA resin chromatography and analyzed by SDS-PAGE and Western blot analysis. The molecular weight of the VP1 protein was 94 kDa. The recombinant virus, VP1-HcNPV-1 did not form polyhedral inclusion bodies and expressed VP1 protein with 95 kDa in the infected S. frugiperda cells, which was detected by Western blot. The titer of the VP1-HcNPV-1 in the first infected cells was $2.0{\times}10^5\;pfu/ml$ at 7 days postinfection.

• Journal of Korean Society of Forest Science
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• v.39 no.1
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• pp.47-56
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• 1978

This study aimed to know difference in freezing resistance among different clonal seedlings or different seed source seedlings of Cryptomeria japonica which has been selected where extreme cold prevails in Korea and Japan. The freezing resistance of three 12-50 year old trees was also measured in the experiment. The freezing resistance was measured in different tissue parts: mainly leaf, cambiam and xylem, at three different collection dates in two different collection places during the winter of 1977-1978. The following results and discussions were made: 1. The clonal difference in freezing resistance of Cryptomeria japonica was $9^{\circ}$ to $15^{\circ}C$ in maximum according to the collection place. However, the clonal difference in freezing resistance was not related to the difference in climatic conditions where the parent tree have been growing. This impiled that the natural selection of cold resistant genes in Cryptomeria japonica has not reached its evolutional equilibrium yet since most of the Cryptomeria forest has been established by artificial regeneration. 2. The difference in freezing resistance among leaf, cambium and xylem was not apparent except that leaf of several clones showed higher freezing resistance than cambium or xylem when they collected at mid-winter. The least freezing resistant tissue part, thought its freezing resistance was not measure in all clones and all temperatures were appeared in the apical buds. The new shoot growth was observed in the next spring with being replaced by its dormant or adventitious bud growth when the apical bud was injured dy cold during winter. 3. The freezing resistance of leaf, cambium and xylem was shown high enough so that freezing resistance Cryptomeria clones in this experiment were supposed to be able to survive in cold winter conditions at the middle part of Korea. However, it was reported that the most susceptible tissue part to winter injury was the basal stem, but of which freezing resistance was not-measured in this experiment. Several silvicultural methods for prevention of Cryptomeria seedlings from cold damage were discussed in literature.

• Journal of the Korean Society of Food Science and Nutrition
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• v.41 no.8
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• pp.1066-1071
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• 2012

In this study we investigated the change in antigenicity and allergenicity of Meju, a traditional Korean soybean product, by fermentation via 3 different microorganisms. The steamed soybeans were fermented with Lactococcus lactis subsp. lactis and/or Aspergillus oryzae and/or Bacillus subtilis. Proteins in soybean were degraded after fermentation. Antigenicity or allergenicity were analysed by immunoblotting and ELISA using soybean protein-specific polyclonal antibodies or soybean allergic patient sera. The best degradation was achieved by three step fermentation using nisin-producing Lactococcus lactis subsp. lactis IFO12007, A. oryzae and B. subtilis. Allergenicity and antigenicity were also starkly reduced after three step fermentation. The three-step fermentation method developed in our lab suggests an excellent alternative to reduce the allergenicity of soybeans.

• Horticultural Science & Technology
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• v.31 no.2
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• pp.246-254
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• 2013

Five isolates of Tomato yellow leaf curl virus (TYLCV) collected from various regions of Korea were amplified using PCR and determined the sequences of full-length genome, respectively. The PCR-amplified DNA of each TYLCV isolate was introduced into a binary vector to construct infectious clone containing 1.9 copies of the corresponding viral genome. Various cultivars and breeding lines of tomato were inoculated with Agrobacterium tumefaciens harboring infectious clone of each TYLCV isolate to assess resistance against TYLCV. Susceptible cultivar 'Super-sunread' revealed typical yellowing and narrowing of the upper leaves. In contrast, breeding linesTY12, GC9, GC171, and GC173, which contained the TY-1 and/or TY-3 genes that confer resistance against TYLCV in nature, were completely symptomless, suggesting that the lines were resistant to challenging TYLCV isolates. Symptoms of TYLCV in susceptible tomato cultivars are significantly different from those of TYLCV in the resistant tomato cultivars at 30 days after agroinfiltration. Although genomic DNAs of TYLCV were detected from the breeding lines TY12, GC9, GC171, and GC173 using real-time PCR analysis with specific primers, levels of TYLCV DNA accumulation in the resistant breeding lines were much lower than those of TYLCV DNA accumulation in susceptible tomato cultivars. Similar symptom severity and levels of TYLCV DNA accumulation were observed from TYLCV infections mediated by Bemisia tabaci in the resistant and susceptible tomato cultivars. Concentration of agrobacterium did not affect the response of tomato cultivars against TYLCV inoculation. Taken together, these results suggest that TYLCV inoculation via agroinfiltration is as effective as inoculation through Bemisia tabaci and is useful for breeding programs of TYLCV-resistant tomato.

• Journal of Marine Bioscience and Biotechnology
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• v.2 no.3
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• pp.192-197
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• 2007

Suppression subtractive hybridization (SSH) was performed to construct an cDNA library of olive flounder Paralichthys olivaceus peripheral blood leukocytes (PBL) stimulated with lipopolysaccharide (LPS). Total 470 clones were randomly selected and sequenced, 214 out of 470 sequences showed identities with known genes and 252 sequences were unknown genes. Among the 218 known genes, 34 sequences were found to be homologous to IL-1RII gene, and 14 sequences were identified as IL-$1{\beta}$. RT-PCR analysis showed that IL-$1{\beta}$ mRNA was induced 1h after LPS-stimulation, and IL-1RII was increased from 3h after stimulation, indicating that most of SSH clones are associated with inflammatory responses in fish, and this SSH cDNA library would be useful to identify bio-defense and immuno-related genes in fish.

• Korean Journal of Poultry Science
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• v.38 no.4
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• pp.323-330
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• 2011

The chicken monoclonal antibody (mAb), 13C8, reacts with sporozoite antigens of Eimeria spp. which causes avian coccidiosis. Since this mAb was produced at low amount due to genetic instability of chicken hybridoma, a recombinant 13C8 single-chain Fv (ScFv) antibody was constructed by amplification of the variable domain of heavy (VH) and light chain (VL) genes of antibody derived from chicken hybridoma. The constructed 13C8 ScFv was successfully expressed in E. coli and purified as a soluble form. In ELISA analysis, this recombinant 13C8 ScFv antibody showed antigen binding activity as the original mAb. In addition, nucleotide sequence comparison of 13C8 gene to the germline chicken VL and VH genes suggested that the gene conversion with $V{\lambda}$ and VH pseudogenes might contribute to the diversification of VL and VH genes in chickens.

• Journal of Life Science
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• v.17 no.9 s.89
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• pp.1197-1203
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• 2007

A genomic DNA library of the bacterium Paenibacillus sp. DG-22 was constructed and the ${\beta}-xylosi-dase-positive$ clones were identified using the fluorogenic substrate $4-methylumbelliferyl-{\beta}-D-xylopyr-anoside$ $({\beta}MUX)$. A recombinant plasmid was isolated from the clone and 4.3-kb inserted DNA was sequenced. The ${\beta}-xylosidase$ gene (xylA) was comprised of a 2,106 bp open reading frame (ORF) en-coding 701 amino acids with a molecular weight of 78,710 dalton and a pI of 5.0. The deduced amino acid sequence of the xylA gene product had significant similarity with ${\beta}-xylosidases$ classified into family 52 of glycosyl hydrolases. The xylA gene was subcloned into the pQE60 expression vector to fuse with six histidine-tag. The recombinant ${\beta}-xylosidase$ $(XylA-H_6)$ was purified to homogeneity by heat-treatment and immobilized metal affinity chromatography. The pH and temperature optima of the $XylA-H_6$ enzyme were pH 5.5-6.0 and $60^{\circ}C$, respectively.

• Journal of Animal Science and Technology
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• v.44 no.1
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• pp.39-44
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• 2002

The current study was conducted to investigate the effects of administration of antiserum against adipocyte plasma membrane(APM) proteins into rats on body fat mass. Twenty(20) male adult Sprague-Dawley rats were randomly allocated into either control or antiserum treatment group(10 rats/treatment) and immunized with physiological saline(control group) and polyclonal antiserum (treatment group), respectively, raised in sheep against rat APM proteins(5times, 2day interval). All animals were killed 4weeks after last injection. Intraperitoneal(i.p.) administration of antiserum significantly(P=0.0054 and P=0.0019, respectively) reduced subcutaneous(21.9%) and perirenal + mesentric + epididymic(36.0%) adipose tissue mass in rats of treatment group. Although body weights of antiserum treated rats were decreased during immunization, the rats recovered their body weight after 1 week of treatment. There were no significant changes in the level of blood glucose and in the contents of muscle protein and fat in antiserum treated animals. Current results indicate that polyclonal antibodies against APM proteins could be used to manipulate body fat mass in meat animals as well as laboratory animals. Further studies, however, are necessary for the practical applications of the current results.

• Korean Journal of Food Science and Technology
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• v.35 no.3
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• pp.366-372
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• 2003

Enzyme-linked immunosorbent assay (ELISA) for assaying the 5-enolpyruvyshikimate-3-phosphate synthase from Agribacterium sp. CP4 (CP4 EPSPS) in genetically modified soybeans was developed. Polyclonal and monoclonal antibodies (Pab, Mab) specific to the CP4 EPSPS were produced. When using the Pab, the detection limit of sandwich ELISA toward CP4 EPSPS (0.03 ${\mu}g/mL$) was better than that of competitive indirect ELISA(ciELISA) (1 ${\mu}g/mL$). It was found that 2 of 3 monoclonal antibodies, Mab1 and Mab2, recognized the same antigenic determinant on CP4 EPSPS, but Mab3 recognized different antigenic determinant when competitive ELISA was performed using the Mabs. On the other hand, when the sensitivity of sandwich ELISA using combination of Pab and/or Mabs was determined, the sandiwich ELISA using Mab2 as a capture antibody and Pab-HRP as a secondary antibody showed the lowest detection limit of CP4 EPSPS (0.02 ${\mu}g/mL$). The sandwich ELISA developed in this study could be applied to detect glyphosate-tolerant soybeans.

• Journal of Korean Society of Forest Science
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• v.99 no.3
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• pp.344-352
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• 2010

To investigate the mating system of clones in the seed orchard of Japanese red pine, parameters of mating system, such as outcrossing rates, number of potential pollen contributors, and degree of pollen contamination, were estimated in the seed orchard of Japanese red pines on the basis of DNA data including 4 nSSR and 6 cpSSR markers. Estimates of outcrossing rates were ranged from 94.9 to 100% with an average of 98.9% on the basis of the analysis of cpSSR haplotypes. They were ranged from 90.3% to 100% with an average of 95.9% on the basis of the analysis of nSSR genotypes. However, cross checking of both DNA markers revealed that the seeds presumed to be products of self pollination were turned out to be generated by pollination between mother tree and other tree (i.e., 100% of cumulative outcrossing rate). Estimates of pollen contamination ranged from 43.6% (Gangwon-10) to 56.4% (Gangwon-12) with the average of 48.9%. On the basis of pooled cpSSR haplotype of each seed, maximum number of 21 pollen contributors were verified from the seeds reproduced by Kyungbuk-38. Minimum number of 13 pollen contributors were verified in Gangwon-10. Mean of 16.2 pollen contributors were verified from a total of 5 mother trees. In conclusion, considering pretty high outcrossing rates between clones within a seed orchard, it may be expected that a fairly good genetic potential of the seeds, produced in '77 plot of the seed orchard of Japanese red pines at Anmyeon island, may be guaranteed. Observed results from the analysis of mating system of Japanese red pines in a '77 plot of the seed orchard may also provide useful information for the establishment and management of the seed orchard of the progressive generation.