• Korean Journal of Animal Reproduction
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• v.17 no.1
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• pp.43-48
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• 1993

These experiments were carried out to construct mouse testis cDNA library and to to seen H-Y Ag gene. Mouse testis was obtained from BALB/c inbreed mouse that was after-born 1 week. Isolation of mouse testis total RNA was carried out by guanidum/cesium choloride, poly(A+) mRNAs were purified by oligo d(T)-cellulose chromatography method. To investigate protein synthesis activity, in-vitro translation carried out by total RNA and poly(A+) mRNA. The products of in-vitro translation were identified in 12.5% PAGE. Single strand DNA and double strand DNA were synthesized from poly(A+) mRNA and purified using phenol/chloroform/isoamylalcohol. Synthesized cDNA was combined with cohesive Eco RI polylinker, its recombination efficiencies were identified by X-gal and IPTG. In the cDNA library, 1$\times$107 phagemids were screened with 32P labelled probe. Hybridization were carried on $65^{\circ}C$ for 16~20hours. And 1$\times$106 phagemids were screened with rabbit-anti-H-Y. In former, select 5 positive clones, and later, 1 positive clone. Its southern blot analysis showed various size of insert cDNA from 0.7kb to 3kb.

• Applied Microscopy
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• v.19 no.2
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• pp.74-84
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• 1989

Buffer soluble storage proteins of ginseng seed have been localized by electron microscopy using post-embedding immunocytochemical gold labelling technique. Major components of the storage proteins were revealed to be storage protein-1($SP_{1}$, MW 160,000) and storage protein-2($SP_{2}$, MW 70,000). Both of the storage proteins are glycoproteins. Anti-$SP_{1}$ and anti-$SP_{2}$ from rabbit, against $SP_1$ and $SP_2$, respectively, reacted on sections of ginseng endosperm tissue embedded in Spurr's epoxy resin. The rabbit antibodies were visualized indirectly by reaction with protein A labelled with colloidal gold. Both storage proteins were found to be accumulated together in the same protein bodies, but their relative contents are not equal.

• Korean Journal of Veterinary Service
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• v.13 no.1
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• pp.32-43
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• 1990

In order to investigate the clinico-pathological and immunohistochemical changes in the rats infected with Aujeszky’s disease virus(ADV), 100 heads of 4 weeks-old rats were inoculated intraperitoneally and intranasally, with the domestically isolated ADV, NYJ-1-87 strain, at $10^{3.0}$ or $10^{5.0}$$TCID_ {50}$/0.2ml. Results obtained through the experiments were summarized as follows : 1. Clinical signs such as dulness, anorexia, pruritus, fascial edema, dyspnea and ataxia were observed from the 2nd day and died at the 3rd to 5th day after ADV inoculation. By necropsy, congestion and hemorrhage were observed in the abdominal organs, while no specific changes were detected in the other organs. 2. In histopathological observation, degeneration and necrosis of the nervous cells, non-suppurative meningoencephalitis, microgliosis and perivascular cuffing were manifested in central nerve system but no specific changes were observed in the other organs. 3. By immunohistochemical staining using peroxidase antiperoxidase, the positive cells were detected in the tissues of kideny, spleen, urinary bladder and lung.

• Korean Journal of Veterinary Service
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• v.13 no.1
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• pp.54-63
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• 1990

We have studied about the safety, immunity and protective potency in swine and experimental am mais of two inactivated vaccine produced with NYJ-1-87 strain of ADV that was isolated in Korea. Result obtained through the experiments were summarized as follows. 1. The safe potency of ADV antigens inactivated with BEI and formaline to mouse & guinea pig was on the whole good, but protective potency rates of those to challenge with ADV were 60-75％ without the differences to two antigens. 2. Safety, immunity & protective potency of ADV antigens inactivated with BEI and formaline to swine were on the whole excellent, except for a mild increase of rectal temperature in some pigs after challenge with ADV. 3. When virus excretion of the experimental groups after challenge with ADV was examed by swabbing of nasal, all pigs of control gorup excreted virus from 2 days p.c., partially to 10 days p.c.. But in vaccinated groups, only 25-50% of all pigs of each group excreted virus during experimental periods. 4. Titers of antibodies in swine & quinea pig vaccinated with inactivated ADV antigens become increased after the 1 weeth p.i. showing the highest liters on the 4-5 weeths p.i.

• Korean Journal of Veterinary Research
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• v.36 no.3
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• pp.693-698
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• 1996

Haemaphysalis longiscornis is the common cattle tick of great economic importance in Korea. Chemical control using dips or sprays has been the traditional method of attempting to kill these ticks during the infestation period. However, the presence of resistant forms to chemical, the rising costs of acaricides and environmental problems have made it almost impossible to use these chemicals on a regular basis according to the pest problem. For this reason, vaccination against ticks and breeding for host resistance against ticks are being studied. In order to determine the common proteins and antigens according to developmental stages, SDS-PAGE and western blotting were performed. In SDS-PAGE 103.3kD and 98.3kD proteins were observed as common proteins, and these proteins were observed as common antigens in western blotting. Unimmunized rabbits were infestated three times with H longicornis. The weight of the second and the third engorged ticks were 0.153g and 0.104g respectively. This weight is 69% and 47% of the first engorged ticks weight respectively. Immunized rabbits by adult ticks antigen and control were infested with H longicornis. The control taked 3-4 days to fully engorge, but the immunized rabbits taked about 7 days. So adult tick antigen may be effective to render the immunity to host.

• Research in Plant Disease
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• v.16 no.2
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• pp.121-124
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• 2010

Immunocapture reverse transcription polymerase chain reaction (IC-RT-PCR) was applied to the detection of Cucumber green mottle mosaic virus (CGMMV), Kyuri green mottle mosaic virus (KGMMV), and Zucchini green mottle mosaic virus (ZGMMV) on Cucurbitaceae crops. These seed-borne tobamoviruses were accurately detected from the infected leaves and seeds by IC-RT-PCR. This method was estimated to be about 100 times more sensitive than ELISA, and also it allowed the direct confirmation of ELISA results by using the captured antigens from a completed ELISA microwell. This convenient and reliable method could be used routinely for large-scale field surveys or seed tests of Cucurbitaceae crops.

• The Journal of Korean Society of Virology
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• v.27 no.1
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• pp.9-17
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• 1997

Hepatitis C virus (HCV) E2 protein is known to be one of putative envelope proteins. To develop a sensitive detection method for HCV infected tissues and cells, monoclonal antibodys (MAbs) to the E2 protein of HCV were prepared from mice immunized with recombinant baculovirus-expressing E2 protein (Bac-E2). Several hybridoma clones secreting various levels of MAb were isolated and isotypes of these MAb were determined. One clone (L.2.3.3) was used for ascites production and the E2-MAb was purified and characterized. The L.2.3.3 reacted well with both Bac-E2 and E. coli expressed glutathione-S-transferase-E2 (GST-E2) fusion proteins. Using HCV patient sera, E2 envelope protein was found to be localized in the cell membrane boundary both in CHO cells and insect cells which express HCV E2 protein. Similar result was obtained when same cells were treated with the MAb L.2.3.3. These results demonstrated that Bac-E2 protein is capable of eliciting high titer antibody production in mice.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1995.04a
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• pp.57-57
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• 1995

1) 효소원의 조제 : 류마티스 관절염환자의 관절액 및 rat platelet PLA$_2$는 장등의 방법으로 조재하여 사용하였으며, 기질은 1-pal- (1-$^{14}$ C) linoleoyl PE로 하여 Dole 변법으로 효소활성을 측정하였다. 2) Histamine 유리반응 ; Rat복강으로부터 정제한 비만세포를 항원-항체 복합체로 자극하거나, $A_{23187}$등의 처리로 유리되는 histamine 을 methylation 시킨 후 유기용매법으로 추출한 후 scintillation counter로 측정하였다. 결과 : \circled1 천연물로부터 분리한 5종의 biflavonoid (amentoflavone 및 그 유도체)의 PLA$_2$ 저해 활성을 검토한 결과 거의 유사한 IC50 (약 3$\mu\textrm{m}$)을 나타내었다. \circled2 이들 중 amentoflavone은 염증성 PLA$_2$(Group II형 PLA$_2$)에 비교적 특이성을 나타내었다. 또한 제해양식은 비경쟁적 이면서 비가역적이였다. \circled3 비만세포에서 histamine 유리 억제반응은 자극제의 종류에 관계없이 억제작용을 나타내었으며, 기존에 임상적으로 사용되고 있는 Tranilast나 DSCG(disodium chromoglycate)에 비하여 10배 이상 강한 histamine 유리 억제작용을 나타내었다.다.

• Korean Journal of Veterinary Research
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• v.33 no.2
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• pp.241-247
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• 1993

Forty day old piglets were intranasally inoculated with 2ml of Aujeszky's disease virus (NYJ-1-87 strain, $10^{7.0}$ $TCID_{50/0.2}ml$), and the viral antigens were detected in nasal and circulating white blood cells for 20 days after inoculation by immunocytochemical method. Antibody titers in the blood were also detected by neutralizing test and Aujeszky's disease serodiagnostic kit(Choong Ang) in this periods. 1. Viral antigens were detected by the immunocytochemical technigue, and positive reactions were observated in nasal cells from the 2nd to the l0th days after inoculation and circulated white blood cells from the 4th to the 12th days after inoculation. 2. In neutralization test antibodies levels showed titers of 2 on the 8th day, 8 on the l5th day, 16 on the 18th day and 32 on the 20th day after inoculation. In serodiagnostic kit test positive reactions were observed after the 15th day after inoculation.

• Korean Journal of Veterinary Research
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• v.33 no.2
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• pp.301-308
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• 1993

Three or 7day old piglets were infected experimentally with different encephalomyocarditis virus isolates to detect the viral antigen by the immunoperoxidase technique and to observe strain difference in their pathogenecity in newborn pigs by comparing clinical signs and pathologic lesions. Clinical signs of the infected pigs were different depending on the virus strain, pig age and infection route. Encephalomyocarditis virus(EMCV) NVSL-PR isolate was more pathogenic than MN-25 and MN-30 isolate. Three day old piglets showed more severe illness than 7 day old piglets. Predominant clinical signs were sudden death without noticeable clinical signs and dyspnea manifested as heavy abdominal breathing. Contact-infection from infected piglets to controls was observed in the oro-nasally infected group but not the intramuscular group. Common necropsy findings of dead piglets in both age groups infected with MN-25 and NVSL-PR were accumulation of excessive fluid in the body cavities and mild to diffuse necrotic areas observed in the hearts and occasionally in the livers. Microscopically, myocarditis with inflammatory cell infiltration, necrosis of the myocardial muscle fibers and occasional mineralization were observed along with interstitial pneumonia and centrolobular necrosis in the liver. Using an immunoperoxidase technique, viral antigen was detected in myocardial muscle fibers of piglets infected with EMCV.