• Nuclear Engineering and Technology
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• 제9권4호
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• pp.203-210
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• 1977

디메틸썰폭사이드를 표지보조제로 사용하여 방사면역측정용 $^{125}$/I 표지인슈린을 효과적으로 만들 수 있었다. 디메틸썰폭사이드 조금을 통상의 표지반응 혼합물에 첨가하여 반응시간을 조절하여 보았더니 표지반응속도는 약간 느렸으나 이렇게 하여 얻은 표지인슈린은 항체와의 결합능이 좋았으며 표준곡선을 점시했을 때 큰 구배를 보였다. 표지반응수율도 반응시간을 1분으로 늘림에 따라 약 30%로 늘어났다. 평균 항체결합능(B/F)의 증가는 일정항체 희석비율하에서 표지보조제없이 표지한 인슈린을 쓴 때가 1.5인데 반해 표지보조제를 쓴 경우는 2.5로서 약 70%의 향상을 보였다. 핵자기공명분광법으로 표지혼합물에 첨가한 디메틸썰폭사이드는 클로라민티에 의해 디메틸썰폰으로 산화된다는 것을 확인하였다. Stagg등의 결과와 대략 같은 이 실험결과를 디메틸썰폭사이드와 인슈린분자 중의 유황결합이 클로라민티에 의해 경쟁적 산화를 받는 것으로 보고 이를 토의하였다.

• 한국식물병리학회지
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• 제2권1호
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• pp.48-52
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• 1986

흰토끼풀에 황색모자이크병징을 나타내는 이병주를 채집하여 병즙액의 지표식물반응과 물리적 성질조사, 풍매전염시험, 바이러스입자의 형태관찰 그리고 항혈청반응 등의 방법으로 병원바이러스의 동정을 시도한 결과 강낭콩황반모자이크바이러스(BYMV)로 밝혀졌다. 공시병원바이러스의 지표식물반응은 명아주에 전체국부반응, 잠두의 접종엽에 국부반응, 줄기와 상위엽에 각각 양저증상과 모자이크반응을 보였으며, 강남콩에는 엽맥양저, 황색모자이크 또는 축엽이 되고 순이 고사하였다. 또한 완두, 녹두, 팥과 red clover에는 모자이크 반응을 나타냈다. 바이러스입자의 형태 및 크기는 740nm정도의 사상입자이며, 복숭아혹진딧물에 의하여 전염이 가능하였다. 바이러스병즙액의 생리적 성질은 내열성이 $60\~65^{\circ}C$, 내희석성이 $10^{-3}\;-\;10^{-4}$, 내보존성이 3일이었다. 순화된 공시바이러스는 일본 식물바이러스 연구소로부터 분양받은 BYMV항혈청과의 혼합침강반응결과 양성반응을 보였다.

• 대한미생물학회지
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• 제16권1호
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• pp.39-48
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• 1981

보체결합반응은 감염초기의 항체검출에 대한 민감성이 높기 때문에 사람과 동물의 만성질환, 특히 매독, 브루셀라병. 결핵병, �냑鵝뵉퓌鑿�, 클래미디아감염증, 마이코플라스마감염증, 콕시오이드병, 히스토플라스마병 등의 면역학적진단수기와 예방관리를 위한 전염병학적연구목적에 흔히 사용된다. 그러나 검사혈청이 항보체성 또는 친보체성을 갖는 경우는 특이항체를 검출할 수 없는 단점을 갖고 있다. 그러므로 이 연구에서는 친보체성분을 갖는 돼지혈청을 항보체성 사람, 토끼 및 소혈청에 보강하여 특이 항체검출을 위한 민감성과 특이성에 관한 기초실험을 실시하였다. 결핵에 감염된 사람, 토끼 및 소혈청중에 높은 항보체작용이 있는 혈청에 친보체성 돼지혈청을 보강할 때 결핵균(다당체 및 단백질항원)에 대한 특이항체가 검출되었다.

• Asian-Australasian Journal of Animal Sciences
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• 제15권12호
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• pp.1708-1713
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• 2002

Endometrial explants obtained from does between days 13 and 21 of pregnancy were cultured in a modified minimum essential medium in the presence of [$^35S$]methionine and [$^3H$]-leucine. Proteins synthesized and secreted into medium were analyzed by fluorography of two-dimensional polyacrylamide gel electrophoresis and fluorography. No marked qualitative changes in patterns of protein production by caprine endometrium between days 13-21 of pregnancy. At least 11 proteins showed consistently a clear spot or a grouping of spots with characteristic location on two-dimensional gels. A major low molecular weight protein consisted of two major isoforms (pI 5.3-6.0) of similar molecular mass (21 kDa). Limited N-terminal sequence analysis of these two isoforms showed that the protein had complete homology with bovine placental and plasma retinol-binding protein (RBP) over the first 20 amino acids. Through use of the antiserum raised against bovine placental RBP, immunoreactive RBP was detected in cultures conditioned by uterine explants prepared at days 13, 15 and 21 of pregnancy. In the present study, proteins synthesized and secreted by caprine endometrium during periattachment period of early pregnancy were characterized. The pregnant endometrium secreted a number of neutral-to-acidic proteins which constituted, in part, the histotroph. A vitamin A-transport protein, RBP, was identified in cultures conditioned by endometrium of days 13-21 of pregnancy. The uterine endometrium is the only source of retinol for embryonic tissues. The uterine RBP appears to transport retinol locally toward embryonic tissues. Secretion of RBP by caprine endometrium of days 13, 15 and 21 of pregnancy suggested that retinol played an important role in conceptus development during periattachment period of early pregnancy.

• Asian-Australasian Journal of Animal Sciences
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• 제22권2호
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• pp.289-295
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• 2009

To develop a safer yogurt for immuno-compromised or allergy patients and to extend shelf-life, a plain yogurt was irradiated with doses of 0, 1, 3, 5, and 10 kGy using a gamma ray and the chemical and microbiological quality and allergenicity change were investigated. There was no difference in the content of protein, total solid, and amino acids of the plain yogurt by irradiation treatment and different storage temperatures (4, 20, and $35^{\circ}C$). The lactic acid bacterial counts of irradiated plain yogurt had approximately 3-decimal reduction at 3 kGy, and no viable cell at 10 kGy regardless of storage time and temperature. The binding ability of rabbit antiserum to milk proteins in irradiated plain yogurt showed that 10 kGy of irradiation produced significantly higher binding ability than other treatments. Sensory evaluation indicated that only appearance of the plain yogurt irradiated at 3 kGy or higher had a lower value than the non-irradiated control when stored at $20^{\circ}C$. Results suggest that irradiation of plain yogurt does not significantly affect the chemical and sensory quality of plain yogurt, but can extend the shelf-life, possibly reduce allergenicity, and provide a safer product.

• 한국수산과학회지
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• 제51권3호
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• pp.254-261
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• 2018

From 2014 to 2017, mortalities of seawater-cultured rainbow trout Oncorhynchus mykiss, were observed in the Goheung and Jeju areas of Korea, with Vibrio anguillarum (seven strains: RT1, 2, 3, 4, 5, 6, and 7) identified as the etiological agent. The phenotypic (based on API 20NE, API ZYM, and E-test kits), serotypic (slide agglutination tests with O1, O2, O3, O4, and O7 antisera), and genotypic (16S rRNA and ompU sequencing) characteristics of the seven RT strains were analyzed and compared to those of seven additional V. anguillarum stains (SF, isolated from sweet fish; FM, isolated from flathead mullet; ATCC43305; ATCC43311; ATCC43307; ATCC43308; and KCTC2711). The phenotypes of the RT strains showed variance, while the slide agglutination tests of the RT1-7, SF, and FM strains all showed positive reactions with serotype O1 antiserum. The 16S rRNA and ompU sequences of the RT1-7, SF, and FM strains were affiliated with V. anguillarum ATCC43305 (Serotype O1), but the ompU sequence of the SF strain differed from those of the RT1-7, FM, and ATCC43311 strains, including one amino acid substitution. We thus confirmed that serotype O1 V. anguillarum, with multiple phenotypes, continues to infect seawater-cultured rainbow trout in Korea.

• Journal of Microbiology and Biotechnology
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• 제9권6호
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• pp.778-783
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• 1999

Campylobacter is a pathogen for both humans and animals that can be transferred from animals to humans. Four isolates, which grew under 5-10% $CO_2$ and had small and translucent colonies, were obtained from swine gastric mucosa and characterized using various methods. These bacteria were gram negative, spirally shaped with round ends. One or two non-sheathed polar flagella were observed under electron microscopy. A PCR with species-specific protein (SSP) primers for 16S rRNA gene in Campylobacter produced a typical 462 bp fragment. The isolates had various biochemical and molecular characteristics which differentiated them from other Campylobacters. The isolates were catalase and oxidase positive, urease (rapid) negative, nitrate reduction positive, indoxyl acetate hydrolysis positive, y-glutamyl transpeptidase negative, and alkaline phosphatase negative. All four isolates showed growth at $37^{\circ}C{\;}and{\;}42^{\circ}C{\;}but{\;}not{\;}at{\;}25^{\circ}C$, were resistant to cephalotin and cefoperazone, and susceptible to carbenicillin. The isolates showed various results in the reduction of chloride to triphenyl tetrazolium (TTC) and a susceptibility to nalidixic acid. Western blot analysis of these isolates with antiserum raised against one isolate showed different patterns from those of reference strains. A dendrogram drawn with the RAPD results showed that these isolates belonged to a new Campylobacter spp. group different from those of C. jejuni, C. doylei, C. lari, and C. coli.

• Asian-Australasian Journal of Animal Sciences
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• 제15권6호
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• pp.783-788
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• 2002

Bovine yolk sac at day 24 of pregnancy, and placental membranes (chorion and allantois) from days 70 and 100 of pregnancy were isolated and cultured in a modified minimum essential medium in the presence of $[^{35}S]$methionine. Proteins synthesized and secreted by isolated bovine yolk sac, chorion and allantois were analyzed by fluorography of two-dimensional polyacrylamide gel electrophoresis. Serum-like proteins,transferrin, ${\alpha}$-fetoprotein, ${\alpha}$1-antitrypsin and ${\alpha}$1-acid glycoprotein,were the major protein products of yolk sac. A 21 kDa protein produced by yolk sac was identified immunochemically as retinol-binding protein (RBP). Chorion and allantios from days 70 and 100 of pregnancy were active in protein synthesis and secretion. Both chorion and allantois did not secret serum-like proteins but secreted a number of neutral-to-acidic proteins including RBP. Secretory proteins produced by the yolk sac, chorion and allantois may play important roles in the embryonic development and the successful outcome of pregnancy. Antiserum against bovine placental RBP was employed to the immunocytochemistry by immunoperoxidase method. Immunoreactive RBP was localized in epithelial cells and island-like cell clones of yolk sac. Immunostaining for RBP was detected in simple columnar epithelium of chorion and in simple squamous epithelium of allantois. In the present study, proteins synthesized and secreted by yolk sac at day 24 of pregnancy, chorion and allantois from days 70 and 100 of pregnancy were characterized In addition, RBP was localized in yolk sac, chorion and allantois by immunoperoxidase method. The immunoperoxidase method has been proven to be a very effective technique to identify the cellular source of protein synthesis in extraembryonic membranes.

• Journal of Microbiology and Biotechnology
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• 제19권1호
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• pp.108-112
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• 2009

A simplified method for the purification of cholera toxin was developed. The 569B strain of Vibrio cholerae, a recognized hyper-producer of cholera toxin, was propagated in a bioreactor under conditions that promote the production of the toxin. The toxin was separated from the bacterial cells using 0.2-${\mu}m$ crossflow microfiltration, the clarified toxin was passed through the membrane into the permeate, and the bacterial cells were retained in the retentate. The 0.2-${\mu}m$ permeate was then concentrated 3-fold and diafiltered against 10 mM phosphate buffer, pH 7.6, using 30-kDa crossflow ultrafiltration. The concentrated toxin was loaded onto a cation exchange column, the toxin was bound to the column, and most of the impurities were passed unimpeded through the column. The toxin was eluted with a salt gradient of phosphate buffer, pH 7.0, containing 1.0 M NaCl. The peak containing the toxin was assayed for cholera toxin and protein and the purity was determined to be 92%. The toxin peak had a low endotoxin level of $3.1\;EU/{\mu}g$ of toxin. The purified toxin was used to prepare antiserum against whole toxin, which was used in a $G_{M1}$ ganglioside-binding ELISA to determine residual levels of toxin in an oral inactivated whole-cell cholera vaccine. The $G_{M1}$ ganglioside-binding ELISA was shown to be very sensitive and capable of detecting as little as 1 ng/ml of cholera toxin.

• Parasites, Hosts and Diseases
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• 제31권1호
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• pp.37-42
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• 1993

질트리코모나스(Trichomonasvqqin is)의 항원성 변이를 관찰하기 위해 원충의 표면 항원 (surface antigen)을 N-hydmwsuccinlmide-biotin(N반5-biotin)으로 표지 하고 표지된 표면 단백질과 토끼의 항혈청으로 면역침전(Immunoprecipitation; IP)시켰으며 sodium dodec낀 sulfate-polyacrylamide gel electrophoresis (SDS-PAGE)와 전기영동이적법을 시 행하였다. 살아있는 원충을 NHS-biotin으로 표지하여 표면 단백질을 분리하고 이를 질트리코모나스에 면역된 토끼 항혈청과 면역침전 시켰던 바 46, 60, 68, 90, 130 그리고 220 kDa에서 6개의 단백질이 항원성을 나타내었으며 질토리코모나스의 분리주인 HY-1, HV-15 및 ATCC 50148 주간에 차이는 없어 이들 6개 분획이 표면 항원성 발현에 중요한 역할을 할 것으로 생각된다.