• 생명과학회지
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• 제20권9호
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• pp.1294-1298
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• 2010

흰반점바이러스(WSSV)는 새우양식산업에 막대한 경제적 손실을 일으키는 가장 치명적인 바이러스성 질병 원인체 중 하나이다. 본 연구는 인위적인 실험모델로서 WSSV에 대한 자주새우(Crangon affinis)의 감수성을 확인하고자 실험을 수행하였다. WSSV가 희석된 수조 내에서 침지법으로 감염된 새우는 감염 후 7일째에 100% 누적폐사율을 보여 자주새우가 WSSV에 대해 매우 높은 감수성을 갖고 있음을 확인하였다. 또한 재조합단백질인 rVP466에 대한 항혈청의 중화효과를 확인하기 위해 항혈청과 반응시킨 바이러스액($1{\times}10^4$ 배로 희석된 WSSV)을 이용하여 침지법으로 자주새우에 대해 공격실험(challenge test)을 수행하였다. 실험 결과, WSSV로 challenge한 감염대조구(positive control)의 새우들은 감염 후 5일째에 100% 폐사하였으며, WSSV와 rVP466 항혈청을 1:0.01, 1:0.1, 1:1로 혼합한 액으로 challenge한 새우들은 감염 후 14일째에 각각 100%, 68.8%, 68.8%의 누적폐사율을 보였다. 따라서 본 연구 결과는 연안서식종인 자주새우가 양식장으로부터 배출된 WSSV에 의해 자연상태에서 감염 될 수 있는 가능성과 함께 항혈청에 대한 중화효과를 나타냄으로써 겨울철 저수온기에 WSSV 감염을 위한 대체 실험생물로서의 유용성을 확인하였다.

• 대한수의학회지
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• 제16권1호
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• pp.7-10
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• 1976

An agglutinability between naturally infected positive chicken serum of pullorum disease and hyperimmumized rabbit antiserum was compared. And the following results were obtained and summarized. 1. On the agglutinability, Salmonella pullorum antigen which irradiated gamma-ray was more excellent than another both formalized and heated antigen. 2. Time of judgemented as positive titer in the tube agglutination test to the naturally infected positive chicken serum was it most suitable for 12 hours at $37^{\circ}C$. 3. Agglutination titer of positive immune chicken serum against gamma-ray irradiate Salmonella pullorum were as 320~640x.

• Journal of Plant Biotechnology
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• 제4권1호
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• pp.39-44
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• 2002

The fast-migrating cationic peroxidase isozyme, named RC3, was purified from rice (Oryza sativa cv. Nak-Dong) callus. Purification of the enzyme was accomplished by ammonium sulfate fractionation, CM-cellulose ionexchange chromatography, and Sephacryl S-100 gel filtration. The molecular mass of the enzyme was about 34 KDa as determined by SDS-PACE and 38 KDa by Sephacryl-100 gel filtration. The pI value of the enzyme was 8.9. Antiserum against RC3 was raised in rabbits, and anti RC3 antiserum reacted with RC3 isozyme by Ouchterlony double immunodiffusion. The optimum pHs and Km values of the enzyme for various substrates were determined. Kinetic studies with various substrates showed that RC3 had very low Km value of 0.01 mM for ferulic acid and ascorbic acid. However, the enzyme did not use esculetin as a substrate.

• 한국가축번식학회지
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• 제10권1호
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• pp.36-41
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• 1986

These experiments were carried out to obtain basic information necessary for sexing embryos by chromosomal analysis. To observe metaphase chromosomes, all embryos developed to blastocysts were cultured in Ho, pp. & Pitts' medium containing 0.001% Colcemid under the gas phase of 5% CO2 in air at 37$^{\circ}C$ for 2 hours. The sex chromosome of mouse embryos shown normal development after culture in medium containing H-Y antiserum (10%, v/v) and complement (20%, v/v) also was confimed by chromosomal analysis. The results obtained in these experiments were summarized as follows: 1. Among 89 mouse blastocysts, the number of embryos identified to have XX and XY chromosome was 22(25%) and 25(28%), respectively and 42(47%) embryos were not identified. 2. Of total 40 mouse balstocysts cultured in medium containing H-Y antiserum and complement, 23(58%) embryos which were able to be discriminated their sex chromosomes were identified to be XX bearing embryos. 3. Sex chromosomes of a number of embryos subjected to chromosomal analysis were not identified. This result may be due to absence or poor quality of metaphase spreads.

• KSBB Journal
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• 제5권4호
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• pp.315-321
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• 1990

Rabbit anti-bovine heart PDH antiserum was raised against El(a, b) isolated from PDC, and then applied to detect Ela and Elb. Appropriate amounis of El were fractionated by SDS-PAGE and electrophoretically transferred to nitrocellulose membrane. The Ela and Elb on the membrane were incubated with anti-El antiserum and identified by GAR-HRP system. It has been found that the immunodetection sensitivity of Ela and Elb were directly proportional to the amount of antigen and transfer time. The lengthy transfer times increased the immunodetection sensitivity of Ela and Elb. The maximal detection sensitivity of Western blotting of Ela and Elb was achieved at 3.5 V/cm for 16-hour transfer under these experimental conditions.

• International Journal of Industrial Entomology and Biomaterials
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• 제29권2호
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• pp.214-219
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• 2014

We prepared antibodies from insect glycosaminoglycans (GAGs) and assayed the titer. Nine polyclonal antibodies against insect GAGs were raised for development of an ELISA in biological fluids (mice serum). The 3th booster collection of antiserum of BALB/c mice as a primarily antibody was assayed for titer determination by ELISA method. In sandwich ELISA of GAGs derived from Isaria sinclairii or other insects, antiserum from insect GAGs gave satisfactory results for so potent antibody(100: 1~1000:1) raising (manufacturing) agent in range of 10 ng/ml.

• 한국식물병리학회:학술대회논문집
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• 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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• pp.150.2-150
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• 2003

Carnation mottle carmovirus(CarMV) was isolated from Lilium spp. in Korea. This isolate, CarMV, was done bioassay, which plants were Dianthus caryophyllus, Gomphrena globosa, Chenopodium amaranticolor, Dianths chinensis. CarMV was propagated on the leaves of Chenopodium amaranticolor with the crude-sap inoculation method and purified by Mossops method(1976). We produced antiserum against CarMV and analyzed the antiserum specificity with ELISA, Gel diffusion method, and Rapid Immunofilter Paper Assay (RIPA). From these results of the assay, RIPA method was simple and rapid for CarMV detection. We have established successfully the CarMV detection system. CarMV coat protein gene was amplified by RT-PCR with specific primers and sequencing analysis was done.

• 한국식물병리학회:학술대회논문집
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• 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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• pp.127.2-127
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• 2003

YMV-K strains were purified from D. oppostita Thunb. tv. Dung-Gun-Ma showing mosaic symptom on their leaves. YMV-K strains were filamentous particles of 780nm in length and induced cytoplasmic disorder such as inclusion body formation. Nucleotide and amino acid sequences of 5'-UTR, Pl and CP of YMV-K strains shared 80.8, 64.7 and 98.3％ identity respectively to JYMV J1 in the mean value. Purification of YMV-K strains according to JYMV purification method(S. Fuji) was conducted to product antiserum. With antiserum against YMV-K strains, the Various diagnosis methods such as IC-RT-PCR, DIBA, RIPA and indirect-ELISA were used to detect YMV-K strains in Chinese yam plant.

• 한국생명과학회:학술대회논문집
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• 한국생명과학회 2001년도 제34회 학술심포지움
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• pp.34-34
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• 2001

• 한국식품영양과학회지
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• 제27권6호
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• pp.1152-1159
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• 1998

In order to develop more rapid and reproducible analysis of biotin known as vitamin H, attempts were made to establish the condition for enzyme linked immunosorbent assay(ELISA) compared with traditional microbiological assay(MBA). Antibiotin and antiserum were obtained from the immunized rabbits injected with emulsion of biotin KLH conjugate and Freund's adjuvant. The antiserum showed cross reactivity on biocytin, a derivative of biotin, which is converted to biotin in intestine, at the rate of 177%(median inhibitory concentration(IC50)=12.58ppb), but not on other derivatives such as desthiobiotin, diaminobiotin and 2 imino biotin. Specific antibody for biotin was purified from the antiserum through protein A column and desalting column. The conditions of competitive direct ELISA (cdELISA) were established. Detection range of biotin concentration by cdELISA was 0.01∼300ng/ ml(ppb). In the spike test with milk, fruit flake and pine carrot juice, the correlation coefficient between two methods of MBA and ELISA was reliably consistent at the value of r=0.992. But detection of biotin by microbiological assay(MBA) was rather restricted in range and nonspecific. Detection range of biotin by MBA was 0.1∼0.5ng/ml(ppb). It showed cross reactivities on biocytin and desthiobiotin at the rate of 80.1% and 66.7%, respectively. In conclusion, ELISA revealed a significant improvement compared with MBA for the biotin detection in terms of sensitivity, detection range and cross reactivity. In addition, a variety of samples could be analyzed rapidly and conveniently at one time by using ELISA. These results strongly suggest that the ELISA is very promising for the practical application to detect biotin contents in a wide range of food stuffs.