• 한국양식학회지
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• 제10권4호
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• pp.425-433
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• 1997

황해산 두족류 6종(참갑오징어, Sepia esculenta ; 쇠갑오징어, Sepiella japonica : 한치꼴뚜기, Loligo chinensis : 살오징어, Todarodes pacificus : 참꼴뚜기, Loligo beka : 낙지, Octopus minor)의 안구단백질을 추출하여 항원-항체 교차반응을 실시하였다. 쇠갑오징어, 살오징어와 낙지의 안구단백질로 항혈청(antiserum)을 제작하였으며 갑오징어목, 살오징어목 및 낙지목에 속하는 두족류 6종의 안구단백질을 항원(antigen)으로 사용하였다. 갑오징어목 속하는 쇠갑오징어 및 참갑오징어에서 항원-항체 반응 양상이 매우 뚜렷하였으며 종간 비교에 기준이 될 수 있고 또한 유의성있는 침강선을 볼 수 있었다. 또한 gel filtration chromatography 방법으로 살오징어의 안구단백질을 분획하여 crystallin을 분리하였으며, 주 분획을 SDS-PAGE 전기영동 방법으로 분자량을 추정해 본 결과 약 20-35 KDa 였다.

• Restorative Dentistry and Endodontics
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• 제6권1호
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• pp.93-103
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• 1980

Streptococcus mutans were isolated from dental plaques of carious lesions of 4 patients on mitis-salivarius agar medium. Three patients known to harbor S. mutans in their dental plaques. Identification of the isolated S. mutans was established by colonial morphology on mitis-salivarius agar medium, the fermentation of mannitol and sorbitol, and confirmed by agglutinating reaction with home made anti-S. mutans NCTC 10449 (serotype c) antiserum. Of the isolated S. mutans, one strain (P2-1) showed strong agglutinating reaction with antiserum, another strain (P1-2) showed weak agglutinating reaction. P2-1 strongly adhered to the wall of the test tube containing 5% sucrose broth, while p1-2 weakly colonized on the wall of the test tube. Biotyping of the isolated S. mutans based on the fermentation of mannitol, sorbitol, raffinose and melibiose, and the production of ammonia from L-arginine, and the inhibition of acid production by bacitracin. Biochemical characteristics of P2-1 strain correlated with the recognized biotype c, pl-2 strain resembled biotype d of S. mutans.

• 한국미생물·생명공학회지
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• 제24권5호
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• pp.630-635
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• 1996

For a survey of the occurrence of aflatoxin M$_{1}$ (AFM$_{1}$) in domestic cow's milk, we developed an enzyme-linked immunosorbent assay (ELISA) system, and quantitated the toxin in cow's milk. In order to produce specific antibodies AFM, conjugated to bovine serum albumin (AFM$_{1}$-BSA) and Freund's adjuvant were immunized subcutaneously to rabbits. By use of the antiserum showing the highest titer and AFB$_{1}$-HRP conjugate, we established a competitive direct ELISA (cdELISA) for AFM$_{1}$, whose detection limit was 0.003 ppb. The cross-reactivities of the antiserum against aflatoxin M$_{1}$ M$_{2}$, B$_{1}$, B$_{2}$, G$_{1}$, G$_{2}$, B$_{2a}$, and G$_{2a}$, were 100, 29.9, 25.0, 2.7, 13.0, 0.65, 0, and 0%, respectively. When the cdELISA was applied to the cow's milk spiked with AFM$_{1}$ and followed by cleanup with C$_{18}$ cartridge, the mean recovery of the assay was 104% (mean of CV, 6.4%) in the final concentration of 0.01-1 ppb (10-1, 000 ppt). When cow's milk samples gathered from markets and farms were assayed by the cdELISA, the mean concentration and SD of AFM$_{1}$ was 80.4 $\pm$ 55.0 ppt (n=64; range, 5.6-280 ppt).

• The Plant Pathology Journal
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• 제15권2호
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• pp.119-123
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• 1999

Chrysanthemums showing leaf mottling were collected from three southern locations in Korea in 1998. Two kinds of viruses were isolated from the leaves and were identified as tomato aspermy virus ch-TAV) and chrysanthemum virus B (ch-CVB), according to their host range, morphology, intracellular location, agar gel double diffusion test, and double-stranded RNA (dsRNA) analysis. The purified ch-TAV was spherical particles of approximately 29 nm in diameter and ch-CVB was filamentous particles of 685 nm long. Inclusion bodies were not observed in ch-TAV and/or ch-CVB infected chrysanthemum. ch-TAV showed positive serological reaction with TAV antiserum (ATCC-127) but not with CMV-pepper antiserum. In dsRNA analysis, four kinds of viral dsRNA were observed on ch-TAV and one viral dsRNA was shown on ch-CVB. Rate of co-infection with TAV and CVB in commercial chrysanthemums was 20.9%. On the other hand, infection with CVB alone was 97.2%. However, chrysanthemums naturally infected with TAV alone were not found.

• The Plant Pathology Journal
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• 제36권6호
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• pp.651-657
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• 2020

Lotus is one of the most important aquatic vegetables in China. Previously, we detected sweet potato latent virus from lotus (SPLV-lotus) and found that it has highly significant sequence diversity with SPLV-sweet potato isolates (SPLV-sp). Here, we developed serological methods for the detection of SPLV-lotus in Chinese lotus cultivation areas. Based on the high sensitivity of SPLV-lotus coat protein antiserum, rapid, sensitive and large-scale diagnosis methods of enzyme-linked immunosorbent assay (ELISA) and dot blot in lotus planting area were developed. The established ELISA and dot blot diagnostic methods can be used to detect SPLV-lotus from samples successfully. And our results also showed that the SPLV-lotus and sweet potato isolates appeared clearly distinction in serology. Our study provides a high-throughput, sensitive, and rapid diagnostic method based on serology that can detect SPLV on lotus, which is suggested to be included in viral disease management approach due to its good detection level.

• BMB Reports
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• 제29권2호
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• pp.105-110
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• 1996

Polyclonal antiserum R101 against aflatoxin $B_1$ ($AFB_1$) was raised in New Zealand white rabbits after injection of bovine serum albumin-$AFB_1$ conjugate. Competitive ELISA (enzyme linked immuno-sorbent assay) demonstrated that antiserum R101 has the highest binding for $AFB_1$ (50% inhibition at 170 fmol) and aflatoxicol II (50% inhibition at 112 fmol). It also reacts with other aflatoxins such as $AFB_2$, $AFG_1$, $AFG_2$, and aflatoxin metabolites ($AFM_1$, $AFM_2$, $AFP_1$, and $AFQ_1$), but it does not cross-react with $AFG_2a$. Using this antiserum, aflatoxins were quantitated in 100 urine samples of undergraduate students at the College of Pharmacy, Sung Kyun Kwan University, Republic of Korea. By ELISA, $AFB_1$ and its metabolites were detected in human urine samples (N=100, male=89, female=11, ages=20~31 yrs) with a range of 1.4~200.6 ng/kg/day (mean$\pm$SD=$18.11{\pm}33.01\;ng\;AFB_1/kg/day$ in males, $3.82{\pm}2.65\;ng/kg/day$ in females). Assuming that urinary excretion is about 7.6% of $AFB_1$ intake (Groopman et al., 1992), we estimated that Koreans were daily exposed to a total dietary $AFB_1$ of $240.20{\pm}438.67\;ng/kg/day$ in males and $50.35{\pm}29.88\;ng/kg/day$ in females, respectively. When the human monitoring data was applied to a linear regression model of Y=21.67X-10.04 {Y=liver cancer incidence per 100,000, X=Log $AFB_1$ intake (ng/kg/day), r=0.99} developed from previously reported epidemiological data, calculated liver cancer incidences attributed to $AFB_1$ exposure were 41.56/100,000 in males and 26.84/100,000 in females. The incidences were similarly correlated with liver cancer mortality rates of 43.43/100,000 in males and 11.23/100,000 in females in Korea. These results suggest that aflatoxin exposure may be an important risk factor for the high incidence of liver cancer in Korea.

• 대한수의학회지
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• 제8권2호
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• pp.125-146
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• 1968

Throughout the studies the following experimental results were summarized. 1. It was impossible to infect and kill the mice, weighing 10 to 12 gm, by inoculating 0.2ml of virulent Cl. chauvoei, diluted 1 to 10 with physiological saline, via subcutaneous, intramuscular, intraperitoneal or intraveonus, route. 2. The mice which were inoculated in brain with 0.03ml of Cl. chauvoei diluted 1 : 5120 with physiological saline were resulted in all death after infection, but not in case of attenuated strain even in dilution of one to five. 3. Virulent Cl. chauvoei were diluted with each of those of whole blood, erythrocytes and serum of horse, calf, swine, sheep, rabbit, guinea pig, chicken and duck, human plasma and 2% CaCl solution, and inoculated subcutaneously 0.25 to 0.5ml in mice, weighing 12 to 15gm. It was resulted in significant increase in virulence as comparing with the case of physiological saline solution except when horse and pig sera were used. Such a phenomena were not seen in attenuated strain. 4. Virulence of virulent Cl. Chauvoei could be increased significantly in rat, as the procedures used in mice, by suspending in whole blood, erythrocytes, serum, or plasma of various animals, or 2% $CaCl_2$ solution and by inoculating subcutaneously 0.5 to 10ml in rat, weighing 30 to 60 gm, as compared with those of control group which used physiological saline solutionos diluent. 5. Mice resisted 100 and 80 percent against challenge of $10^3$ and $10^4$ M.L.D.. respectively, 24 hours after inoculation of 0.5ml black leg antiserum. 6. Immune response to the black leg living vaccine in mice could be obtained more favorably in the group of respected vaccination rather than those of single inoculation and the most profitable inoculm size of the vacine was 0.5 to 1.0ml. 7. Challenge for the immunized mice could be carried out effectively 3 weeks after first vaccination. 8. Satisfactory results could be obtained by inoculating subcutaneously for the immunization and intracerebrally or subcutaneously for the challenge. 9. Mice which were inoculated with 0.5ml of black leg living vaccine via subtaneucously two times at seven days interval and 21 days after first inoculation and challenged with 5 and 10 M.L.D. of virulent strain, resited 100 and 70 to 80 percent respectively. Same results were obtainable in black leg killed vaccine as the procedures used in living vaccine. 10. There were significantly different resistances against the definite challenge does between the mice groups which were immnuized with the living vaccine diluted five or 10 times and the undiluted. 11. For the biological assay of black leg living vaccine and antiserum, satisfactory results could be obtained using mice.

• 대한수의학회지
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• 제35권4호
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• pp.743-752
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• 1995

The study was carried out to characterize the properties of Korean isolates of EHV from aborted fetuses and determine envelope protein profiles. The results obtained were summarized as follows; 1. Two strains of EHV was isolated from 2 liver samples among 10 aborted fetuses from which the virus isolation was attempted. 2. Morphological and some enzymatic properties of the Korean isolates of EHV which was designated as $LC_1$ and $LC_2$ was identical to those of a reference strain of Australia-N of EHV-1. The Korean isolates of EHV could be propagated on ED cell culture and they formed typical plaques 1 to 2 days after infection in the ED cells from which typical cuboidal particles of 150~170 nm diameter herpesvirus were observed. The virus could be detected specifically from neucleus and cytoplasm of infected cells by flourescent antibody technique using FITC labelled anti-Aust IV(EHV-1) antiserum. The Korean isolates, $LC_1$ and $LC_2$ were specifically neutralized by anti Aust IV antiserum and reacted positively to CELISA. 3. The structural polypeptides of purified enveloped virions of $LC_1$ and $LC_2$ isolates of EHV were determined by SDS-polyacrylamide gel electrophoresis to identify the envelope glycoproteins. $LC_1$ and $LC_2$ strains revealed 14 glycoproteins ranging in molecular weight from 190 kD to 31 kD while 17 structural proteins of Aust IV(EHV-1), of which 14 were identical to those of $LC_1$ and $LC_2$, were identified. Upon immunoblotting by rabbit antiserum against EHV isolates and EHV-1(Aust IV), 4 immunogenic proteins of $LC_1$ and $LC_2$ were 135 kD, 88 kD, 64 kD and 59 kD, of which 135 kD, 88 kD and 64 kD proteins were also found in Aust IV(EHV-1).

• Clinical and Experimental Reproductive Medicine
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• 제24권1호
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• pp.21-26
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• 1997

본 연구는 체외생산된 생쥐 배반포기배의 ICM 세포에서 EGF-R 발현유무를 immunosurgery와 indirect immunofluorescence (간접 면역 형광방법) 을 이용하여 조사하고자 실시하였다. 본 실험에 사용된 ICM 세포는 체외수정 후 96시간째에 회수된 생쥐 배반포기배를 immunosurgery 방법을 이용하여 얻어졌으며, 회수된 ICM세포는 생사유무와 EGF-R 발현유무 조사에 공시 되어졌다. 그 결과를 요약하면 다음과 같다. ICM세포의 회수율은 rabbit anti-mouse serum (antiserum) 과 guinea pig serum (complement) 에 각각 15-30 분과 15-60분 동안 처리 했을 경우 8.0-84.2% 였으며, 또한 처리시간이 각각 30분과 60분일 때 가장 높은 회수율 (84.2%) 을 얻었다. Immunosurgery 후 얻어진 ICM세포의 생존 유무를 조사하기 위해 live/dead염색 방법을 이용하였던 바, 처리된 ICM세포중 93.8-100%의 생존율을 나타내어 회수된 ICM세포는 유해한 영향을 받지 않았다는 것을 알 수 있었다. 또한, 간접면역 형광방법을 이용하여 ICM세포에서 EGF-R가 발현되는 것을 확인 하였다. 따라서, ICM세포에서의 EGF-R의 발현은 인위적으로 첨가된 EGF의 이용가능성을 높임으로서 체외에서의 착상전 배 발달을 증진시킬 수 있을 것으로 사료된다.

• 한국식물병리학회:학술대회논문집
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• 한국식물병리학회 1995년도 Proceedings of special lectures on Molecular Biological Approaches to Plant Disease National Agricultural Science and Technology Institute Suwon, Korea
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• pp.27-53
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• 1995

In plant pathology there is an increasing necessity for improved cytological techniques as basis for the localization of cellular substances within the dynamic fine structure of the host-(plant)-pathogen-interaction. Low temperature (LT) preparation techniques (shock freezing, freeze substitution, LT embedding) are now successfully applied in plant pathology. They are regarded as important tools to stabilize the dynamic plant-pathogen-interaction as it exists under physiological conditions. - The main advantage of LT techniques versus conventional chemical fixation is seen in the maintenance of the hydration shell of molecules and macromolecular structures. This results in an improved fine structural preservation and in a superior retention of the antigenicity of proteins. - A well defined ultrastructure of small, fungal organisms and large biological samples such as plant material and as well as the plant-pathogen (fungus) infection sites are presented. The mesophyll tissue of Arabidopsis thaliana is characterized by homogeneously structured cytoplasm closely attached to the cell wall. From analyses of the compatible interaction between Erysiphe graminis f. sp. hordei on barley (Hordeum vulgare), various steps in the infection sequence can be identified. Infection sites of powdery mildew on primary leaves of barley are analysed with regard to the fine structural preservation of the haustoria. The presentation s focussed on the ultrastructure of the extrahaustorial matrix and the extrahaustorial membrane. - The integration of improved cellular preservation with a molecular analysis of the infected host cell is achieved by the application of secondary probing techniques, i.e. immunocytochemistry. Recent data on the characterization of freeze substituted powdery mildew and urst infected plant tissue by immunogold methodology are described with special emphasis on the localization of THRGP-like (threonine-hydrxyproline-rich glycoprotein) epitopes. Infection sites of powdery mildew on barley, stem rust as well as leaf rust (Puccinia recondita) on primary leaves of wheat were probed with a polyclonal antiserum to maize THRGP. Cross-reactivity with the anti-THRGP antiserum was observed over the extrahaustorial matrix of the both compatible and incompatible plant-pathogen interactions. The highly localized accumulation of THRGP-like epitopes at the extrahaustorial host-pathogen interface suggests the involvement of structural, interfacial proteins during the infection of monocotyledonous plants by obligate, biotrophic fungi.