• Proceedings of the Korea Society of Poultry Science Conference
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• 2005.11a
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• pp.44-52
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• 2005

The study, using sequence analysis and phylogenetic relationship of the fusion protein gene, divided the Korean epizootic isolates of Newcastle disease virus (NDV) into several lineages to determine the molecular epidemiology of the virus. A 695 base pair fragment was amplified by polymerase chain reaction between matrix protein gene and fusion protein gene of 30 Korean NDV isolates, which were isolated from field outbreaks of Newcastle disease between 1949 and 2002. All isolates showed the amino acid sequence 112 R-R-Q/R-K-R116 at the C-terminus of the F2 protein and phenylalanine (F) at the N-terminus of the F1 protein, residue 117. These amino acid sequences were identical to a known virulent motif. The region of the F gene between nucleotides 47 and 435 was compared by phylogenetic analysis. Based on nucleotide sequence, the Korean NDV isolates belonged to genotype III, V, VI and VII corresponding to isolates in 1949, 1982 to 1984, 1988 to 1997, and 1995 to 2002, respectively. These data showed that genotypes of five Korean Newcastle disease epizootics had replaced each other serially (III, V, VI and VII) in chronological order. Further, the five Korean Newcastle disease epizootics were closely related with the Necastle disease panzootics or Newcastle disease epizootics in other countries. Present study showed that the Korean genotype V isolated before 1984 was related with European Newcastle disease epizootics in the 1970s, whereas the Korean genotype VI and VII isolated after 1988 were more closely related with Far East Newcastle disease epizootics, especially Newcastle disease3 epizootics in Japan, Taiwan and China. Since 1988, the genotype VI and VII of Far East origin were dominant in South Korea. That might be due to the increased trade of agricultural products including poultry among Far East Asian countries.

• Journal of Microbiology and Biotechnology
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• v.26 no.1
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• pp.160-170
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• 2016

The increasing spread of antibiotic-resistant pathogens has raised the interest in alternative antimicrobial treatments. In our study, the functionally active gram-negative bacterium bacteriophage CP933 endolysin was produced in Nicotiana benthamiana plants by a combination of transient expression and vacuole targeting strategies, and its antimicrobial activity was investigated. Expression of the cp933 gene in E. coli led to growth inhibition and lysis of the host cells or production of trace amounts of CP933. Cytoplasmic expression of the cp933 gene in plants using Potato virus X-based transient expression vectors (pP2C2S and pGR107) resulted in death of the apical portion of experimental plants. To protect plants against the toxic effects of the CP933 protein, the cp933 coding region was fused at its Nterminus to an N-terminal signal peptide from the potato proteinase inhibitor I to direct CP933 to the delta-type vacuoles. Plants producing the CP933 fusion protein did not exhibit the severe toxic effects seen with the unfused protein and the level of expression was 0.16 mg/g of plant tissue. Antimicrobial assays revealed that, in contrast to gram-negative bacterium E. coli (BL21(DE3)), the gram-positive plant pathogenic bacterium Clavibacter michiganensis was more susceptible to the plant-produced CP933, showing 18% growth inhibition. The results of our experiments demonstrate that the combination of transient expression and protein targeting to the delta vacuoles is a promising approach to produce functionally active proteins that exhibit toxicity when expressed in plant cells.

• The Korean Journal of Mycology
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• v.25 no.2 s.81
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• pp.143-151
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• 1997

Interspecific hybrids between Aspergillus niger and Penicillium notatum (Tyr-), hyperlipolytic enzyme-producing fungi, were obtained by nuclear transfer technique. Optimal conditions for formation of intergeneric hybrids were investigated. Maximum production of protoplasts was obtained by 1% Novozyme 234 at $30^{\circ}C$ for 3 hrs and the most effective osmotic stabilizers for the isolation of protoplasts were 0.6 M KCl. Frequencies of hybrid formation by nuclear transfer were $3.8{\times}10^{-3}{\sim}1.3{\times}10^{-3}$. From the observation of genetic stability, conidial size, DNA content, and nuclear stain, it was suggested that their karyotypes are aneuploid. The hybrids showed $1.2{\sim}1.7$ fold higher lipase activities than parental strains. It was strongly supported by results of this study that nuclear transfer technique is much more efficient in the formation of intergeneric hybrids than protoplast fusion and is very useful for the improvement of strains.

• Parasites, Hosts and Diseases
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• v.29 no.3
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• pp.293-310
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• 1991

Clonorchis sinensis is a common parasite of man in Korea. Researches on the specific antigens of C. sinensis would be valuable not only because those elucidate the molecular characteristics of this fluke but also because it is applicable to immunodiagnosis. Although many monoclonal antibodies have been used in the field of parasite immunology, few articles on monoclonal antibodies against C. sinensis have been published so far. The aim of this study was to analyse C. sinensis antigens recognized by monoclonal antibodies, and to set up ELISA-inhibition test using C. sinensis specific monoclonal antibodies for improved specificity of immunodiagnostic tests. By fusion between spleen cells of the mice immunized with C. sinensis water-soluble crude adult worm antigens and plasmacytoma cells of mouse origin, 29 hybridoma clones secreting anti-C. sinensis monoclonal antibodies were made, and 8 clones among those were found specific. After cell cloning, isotypes of 6 selected specific monoclonal anti- bodies were determined to be IgGl, IgG2b and IgA. Four exposed antigenic determinants of natural infection were recognized by different specific monoclonal antibodies. By enzyme-immunoelectrotransfer blot, 10 KD, 34 KD antigenic determinants were found to be reacted with CsHyb 0714-20, CsHyb 0605-10 monoclonal antibodies, respectively, The antigenic determinant recognized by CsHyb 0714-20 monoclonal antibody was revealed to be located at the surface and parenchyme of a parasite by indirect immunoauorescent antibody technique, and those reacted with CsHyb 0605-10, CsHyb 0714-25 monoclonal antibodies were found at the parenchyme and intestine. The antigenic determinant reacted with CsHyb 0605-23 monoclonal antibody was found mainly around the uterine eggs. Four antigenic determinants recognized by specific monoclonal antibodies were all found to be present in the early eluted fractions of C. sinensis antigens separated by Sephadex G-200 gel filtration. By conventional ELISA, 75% of clonorchiasis cases were found positive, but 7.1% of normal controls and 37.5% of paragonimiasis cases showed false positives. However, by ELISA-inhibition test using C. sinensis specific monoclonal antibody (CsHyb 0605-23), 77.1% of clonorchiasis cases were found positive, and there were no false positives in normal controls or paragonimiasis cases, indicating 100% specificity. The ELISA- inhibition test using monoclonal antibodies was found to have same sensitivity and definitely high specificity in comparison with conventional ELISA for serodiagnosis of human clonorchiasis.

• The Journal of Korean Society for Radiation Therapy
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• v.31 no.1
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• pp.25-32
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• 2019

Purpose: The size, shape, and volume of prosthetic appliance depend on the metal artifacts resulting from dental implant during head and neck treatment with radiation. This reduced the accuracy of contouring targets and surrounding normal tissues in radiation treatment plan. Therefore, the purpose of this study is to obtain the images of metal representing the size of tooth through MVCT, SMART-MAR CT and KVCT, evaluate the volumes, apply them into the proton therapy plan, and analyze the difference of dose distribution. Materials and Methods : Metal A ($0.5{\times}0.5{\times}0.5cm$), Metal B ($1{\times}1{\times}1cm$), and Metal C ($1{\times}2{\times}1cm$) similar in size to inlay, crown, and bridge taking the treatments used at the dentist's into account were made with Cerrobend ($9.64g/cm^3$). Metal was placed into the In House Head & Neck Phantom and by using CT Simulator (Discovery CT 590RT, GE, USA) the images of KVCT and SMART-MAR were obtained with slice thickness 1.25 mm. The images of MVCT were obtained in the same way with $RADIXACT^{(R)}$ Series (Accuracy $Precision^{(R)}$, USA). The images of metal obtained through MVCT, SMART-MAR CT, and KVCT were compared in both size of axis X, Y, and Z and volume based on the Autocontour Thresholds Raw Values from the computerized treatment planning equipment Pinnacle (Ver 9.10, Philips, Palo Alto, USA). The proton treatment plan (Ray station 5.1, RaySearch, USA) was set by fusing the contour of metal B ($1{\times}1{\times}1cm$) obtained from the above experiment by each CT into KVCT in order to compare the difference of dose distribution. Result: Referencing the actual sizes, it was appeared: Metal A (MVCT: 1.0 times, SMART-MAR CT: 1.84 times, and KVCT: 1.92 times), Metal B (MVCT: 1.02 times, SMART-MAR CT: 1.47 times, and KVCT: 1.82 times), and Metal C (MVCT: 1.0 times, SMART-MAR CT: 1.46 times, and KVCT: 1.66 times). MVCT was measured most similarly to the actual metal volume. As a result of measurement by applying the volume of metal B into proton treatment plan, the dose of $D_{99%}$ volume was measured as: MVCT: 3094 CcGE, SMART-MAR CT: 2902 CcGE, and KVCT: 2880 CcGE, against the reference 3082 CcGE Conclusion: Overall volume and axes X and Z were most identical to the actual sizes in MVCT and axis Y, which is in the superior-Inferior direction, was regular in length without differences in CT. The best dose distribution was shown in MVCT having similar size, shape, and volume of metal when treating head and neck protons. Thus it is thought that it would be very useful if the contour of prosthetic appliance using MVCT is applied into KVCT for proton treatment plan.

• Journal of Life Science
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• v.17 no.7 s.87
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• pp.889-895
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• 2007

A conventional kinesin, KIF5/kinesin-I, is composed of two kinesin heavy chains (KHCs) and two kinesin light chains (KLCs) and binds directly to microtubules. KIF5 motor mediates the transport of various membranous organelles, but the mechanism how they recognize and bind to a specific cargo has not yet been completely elucidated. Here, we used the yeast two-hybrid system to identify the neuronal protein(s) that interacts with the tetratricopeptide repeats (TRP) of KLCI and found a specific interaction with JNK/stress-activated protein kinase-associated protein 1 (JSAP1/JIPP3). The yeast two-hybrid assay demonstrated that the TRP 1,2 domain-containing region of KLCI mediated binding to the leucine zipper domain of JSAP1. JSAP1 also bound to the TRP region of lac2 but not to neuronal KIF5A, KIF5C and ubiquitous KIF5B in the yeast two-hybrid assay. In addition, these proteins showed specific interactions in the GST pull-down assay and by co-immunoprecipitation. KLCI and KIF5B interacted with GST-ISAP1 fusion proteins, but not with GST alone. An antibody to JSAPI specifically co-immunoprecipitated KIF5s associated with JSAP1 from mouse brain extracts. These results suggest that JSAP1, as KLC1 receptor, is involved in the KIF5 mediated transport.

• Journal of Food Hygiene and Safety
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• v.32 no.1
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• pp.75-81
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• 2017

For people who have a food allergy the only way to manage the allergy is to avoid the food allergen. The mackerel is one of the major food allergens, but no immunoassay for the rapid and simple detection of mackerel has been reported. The objectives of this study are to develop and characterize monoclonal antibodies (MAbs) specific to mackerel using thermal stable-soluble proteins (TSSP) as an immunogen and to characterize the MAbs by indirect enzyme-linked immunosorbent assay (iELISA). The mice immunized with mackerel TSSP and showing high titer were used for cell fusion and cloning. The characterization of MAbs produced from hybridoma cells obtained was confirmed by indirect ELISA and western blot. Four MAbs were confirmed to be specific to mackerel without cross-reaction to other marine products and livestock products in the both methods. The iELISA and western blot based on the MAbs can sensitively detect 1% mackerel protein in other marine products. These results support that immunochemical methods based on the MAb produced could be used as rapid means to detect low levels of mackerel and to identify mackerel adulterated in food.

• IMMUNE NETWORK
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• v.2 no.3
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• pp.175-181
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• 2002

Background: S100A6 is a calcium-binding protein overexpressed in several tumor cell lines including melanoma with high metastatic activity and involved in various cellular processes such as cell division and differentiation. To detect S100A6 protein in patient' samples (ex, blood or tissue), it is essential to produce a monoclonal antibody specific to the protein. Methods: First, cDNA coding for ORF region of human S100A6 gene was amplified and cloned into the expression vector for GST fusion protein. We have produced recombinant S100A6 protein and subsequently, monoclonal antibodies to the protein. The specificity of anti-S100A6 monoclonal antibody was confirmed using recombinant S100A recombinant proteins of other S100A family (GST-S100A1, GST-S100A2 and GST-S100A4) and the cell lysates of several human cell lines. Also, to identify the specific recognition site of the monoclonal antibody, we have performed the immunoblot analysis with serially deleted S100A6 recombinant proteins. Results: GST-S100A6 recombinant protein was induced and purified. And then S100A6 protein excluding GST protein was obtained and monoclonal antibody to the protein was produced. Monoclonal antibody (K02C12-1; patent number, 330311) has no cross-reaction to several other S100 family proteins. It appears that anti-S100A6 monoclonal antibody reacts with the region containing the amino acid sequence from 46 to 61 of S100A6 protein. Conclusion: These data suggest that anti-S100A6 monoclonal antibody produced can be very useful in development of diagnostic system for S100A6 protein.

• Journal of Korean Ophthalmic Optics Society
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• v.18 no.2
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• pp.203-211
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• 2013

Purpose: When we look at the object, we used the dominant eye mainly. For this reason, a prescription of the dominant eye is an important factor for glasses and contact lenses. This study evaluated visual acuity differences between dominant and nondominant eyes through analyzing refractive power changes in both eyes by the ages. Methods: This study was performed to investigate the relationship between refractive error and dominant eye which had the superiority in the function of binocular. 186 subjects without ocular disease were examined on the dominant eye. The dominant eye was examined by the Hole-in-the-card test. For the consistency of the measurements, we tested refractive power in three times by the same person. Results: Using SPSS, the relationship between vision and the dominant eye was analyzed. 135 people of the whole subjects have the dominant eye on right. The Number of the non-dominant eye is 51. We were divided into 3 types, the group under the age of 10 that begins to expose environment factor affect on vision (the average age $8.8{\pm}1.18$) and the age group of 10 to 20 that begins to change refractive power in earnest (the average age $14.1{\pm}2.58$) and the group after the age 20 that began to stabilize vision (the average age $51.8{\pm}17.51$). The visual acuity of dominant eye was higher than non-dominant eye in all age groups. Nevertheless, these results were not statistically significant. Mean astigmatism of dominant eye was smaller than the non-dominant eye, and this is significant, statistically (p=0.017<0.05). Conclusions: It is expected that the balanced eye with a lower level of astigmatism has a more possibility become a dominant eye.

• Nuclear Engineering and Technology
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• v.42 no.6
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• pp.662-669
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• 2010

To develop the manufacturing methods for the blanket first wall (FW) of the International Thermonuclear Experimental Reactor (ITER) and to verify the integrity of the joint, Be/Cu mockups were fabricated and tested at the KoHLT-1 (Korea Heat Load Test facility), a graphite heater facility located at the Korea Atomic Energy Research Institute (KAERI). Since Be and Cu joining is the focus of the present study, the fabricated mockups had a CuCrZr heat sink joined with three Be tiles as an armor material, unlike the original ITER blanket FW, which has a stainless steel structure and coolant tubes. Hot isostatic pressing (HIP) was carried out at $580^{\circ}C$ and 100 MPa for 2 hours as the method for Be/Cu joining. Three interlayers, namely, $1{\mu}mCr/10{\mu}mCu$, $1{\mu}mTi/0.5{\mu}mCr/10{\mu}mCu$, and $5{\mu}mTi/10{\mu}mCu$ were applied as a coating to the Be tiles by a physical vapor deposition (PVD) method. A shear test was performed with the specimens, which were fabricated by the same methods as those used to fabricate the mockups. The average values were 125 MPa to 180 MPa, and the samples with the $1{\mu}mCr/10{\mu}mCu$ interlayer showed the lowest value. No defect or delamination was found in the joints of the mockups by the developed ultrasonic test using a flat-type probe with a 10 MHz frequency and a 0.25 inch diameter. High heat flux (HHF) tests were performed at $1.0\;MW/m^2$ heat flux for each mockup using the given conditions, and the results were analyzed by ANSYS-CFX code. For the test criteria, an expected fatigue lifetime about 1,000 cycles was obtained by analysis with ANSYS-mechanical code. Mockups using the interlayers of $1{\mu}mTi/0.5{\mu}mCr/10{\mu}mCu$ and $5{\mu}mTi/10{\mu}mCu$ survived up to 1,100 cycles over the required number of cycles. However, one of the Be tiles in the other two mockups using the $1{\mu}mCr/10{\mu}mCu$ interlayer was detached during the screening test, and others were detached by discharge after 862 cycles. The integrity of the joints using the proposed interlayers was proven by the HHF test, but the other interlayer requires more study before it can be used for the joining of Be to Cu. Moreover, it was confirmed that the measured temperatures agreed well with the analysis temperatures, which were used to estimate the lifetime and that the developed facility showed its capability of the long time operation.