• 한국식물병리학회:학술대회논문집
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• 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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• pp.133.1-133
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• 2003

Gummy stem blight, caused by Didymella bryoniae occurs exclusively on cucurbits. This fungus has been known not to produce its pycnidium in vitro unless irradiated. Through this study, we optimized cultural conditions for mass-production of pycnidiospore by Metal Halide Lamp irradiation. In brief, the mycelial was cultured at $26^{\circ}C$ on PDA, for 2 days under the darkness, and then the plate was illuminated with MH lamp continuously for 3-4 days at $26^{\circ}C$, a great number of pycnidia was simultaneously formed. Thus produced pycnidiospores were used as immunogen. From fusions of myeloma cell (v-653) with splenocytes from immunifed mice were car ried out. And, two hybridoma cell lines that recognized the immunogen Didymella bryoniae were obtained. One Monoclonal Antibody, Db1, recognized the supernatant and the other monoclonal antibody, Db15, recognized the spore. Two clones were selected which were used to produce ascite fluid two MAb Db1 and Db15, were immunotyped and identified as IgG1 and IgG2b, respectively. Titer of MAb Db1 and MAb Db15 was measured absorbance exceeded 0.5 even at a $10^{-5}$ dilution. The MAbs reacted positively with Didymella bryoniae but none reacted with other of fungi and CMV, CGMMV Sensitivity of MAb was precise enough to detect spore concentration as low as $10^{3}$ well by indirect ELISA characterization of the MAb Db1, Db15 antigen by heat and protease treatments show that the epitope recognized by the MAb Bb1, Db15 were a glycoprotein.

• The Plant Pathology Journal
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• 제19권5호
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• pp.260-265
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• 2003

Gummy stem blight caused by Didymella bryoniae occurs exclusively in cucurbits. This fungus has been known not to produce its pycnidium in vitro unless irradiated. In this study, cultural conditions for the mass-production of pycnidiospore by Metal Halide (MH) lamp irradiation were maximized. The mycelia were cultured at $26^{\circ}C$ on PDA for 2 days under dark condition, and then the plate was illuminated with MH lamp continuously for 3-4 days at $26^{\circ}C$. Results show that a great number of pycnidia were simultaneously formed. The pycnidiospores produced were then used as immunogen. Fusions of myeloma cell (v-653) with splenocytes from immunized mice were carried out. Two hybridoma cell lines that recognized the immunogen D. bryoniae were obtained. One monoclonal antibody (MAb), Dbl, recognized the supernatant while another MAb, Db15, recognized the spore. Two clones were selected which were used to produce ascite fluid of the two MAb, Dbl and Db15, the immunotypes of which were identified as IgG1 and IgG2b, respectively. Titers of MAb Dbl and MAb Db15 were measured and the absorbance exceeded 0.5 even at a $10^{-5}$ dilution. The MAbs reacted positively with D. bryoniae but none reacted with other viral isolates, Cucumber mosaic virus and Cucumber green mottle mosaic virus. Sensitivity of MAb was precise enough to detect spore concentration as low as $10^{-3}$/well by indirect ELISA. Characterization of the MAbs Dbl, Db15 antigen by heat and protease treatments, which suggests that the epitope recognized by these two MAbs was glycoprotein.

• 한국식품위생안전성학회지
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• 제6권3호
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• pp.111-117
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• 1991

가축사료중 zearalenone 분석을 위한 enzyme linked immunosorbent assay(ELISA) 법의 개발을 위해 우선 zearalenone의 항원성을 증폭시키기 위해 zearalenone oxime 유도체를 합성한 다음 bovine serum albumin(BSA)와 conjugate를 만들고, 이를 항원으로 토끼에 면역시켜 11주에 zearalenone에 특이한 항체를 얻어냈다. 생성된 항체를 zearalenone외에 ${\alpha}-zearalenol$과는 강한 cross reactivity를 나타내었고 ${\beta}-zearalenol,\;{\alpha}-zearalenol\;및\;{\beta}-zearalenol$과는 약간의 반응을 보였으며 확립된 ELISA 조건은 당므과 같다. 먼저 시료를 methanol-phosphate buffered saline-dimethyl formate(70 : 29: 1)을 4배 첨가하여 blending 한 다음 Whatman No. 4를 통한 여액을 ELISA시료로 사용하였다. 효소 반응시간과 발색시간은 각각 $37^{\circ}C$에서 30분과 15분이었고, 흡광도는 410nm에서 ELISA reader로서 측정하였으며, 측정한계는 1~100 ppb로 매우 낮았다. 확립된 ELISA 조건으로 실제시료의 zearalenone오염도는 측정결과 24개 시료 중 4개의 시료가 양성반응을 보였고 그 함량범위는 $3.93~7.43\;\mu\textrm{g}/kg$이었다.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2003년도 생물공학의 동향(XIII)
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• pp.186-189
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• 2003

Helicobacter pylori is the etiologic agent of human gastritis and peptic ulceration and produces urease as the major protein component on its surface. H. pylori urease is known to serve as a potent immunogen as well as major virulence factor. In order to express the recombinant urease in tobacco plants, a DNA fragment containing the minimal H. pylori urease gene cluster was subcloned into a plant expression vector. The recombinant vector was transformed to tobacco plants. The integration of the recombinant plasmids into tobacco chromosomal genome was verified by genomic PCR. Expression to mRNA was confirmed by Northern blot analysis, and expression to recombinant urease protein was observed by Western blot analysis. These results showed that the recombinant urease can be produced in tobacco plants and will be tested for immune response to use as an edible vaccine.

• BMB Reports
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• 제31권3호
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• pp.240-244
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• 1998

Helicobacter pylori is the etiologic agent of human gastritis and peptic ulceration and produces urease as the major protein component on its surface. H. pylori urease is known to serve as a major virulence factor and in a potent immunogen. In order to express the recombinant urease at a higher level, a DNA fragment containing the minimal H. pylori urease gene cluster was subcloned into a high copy-number vector. The recombinant H. pylori urease expressed in an E. coli strain that was grown in a rich medium supplemented with added nickel was purified to near homogeneity by using DEAE-Sepharose, Superdex HR200, and Mono-Q (FPLC) columns and the purified enzyme possessed the specific activity of 1255 U/mg. Polyclonal antibodies raised against the purified recombinant H. pylori urease were shown to be very specific when subjected to Western blot analysis, in which crude extracts from the H. pylori ATCC strain and the recombinant E. coli strains expressing various bacterial ureases were exnmined for cross-reactivity.

• 대한약학회:학술대회논문집
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• 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.1
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• pp.290.3-291
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• 2003

To increase detection sensitivity for multi-DDT residues (o,p-/p,p-DDT, o,p-/p,p-DDE, o,o-/o,p-DDD) analysis, a highly selective sample clean-up method was introduced prior to GC/MS analysis using immunoaffinity column. The immunoaffinity matrix was prepared by coupling IgG fraction of DDT antiserum to cyanogens bromide activated Sepharose 4B. Three DDT antisera (DDA-1, DDHP-2, DDCP-3) were test for affinity column ligand that obtained by imunizing respective DDT immunogen to rabbits, and IgG was purified using protein A affinity purification. (omitted)

• Biomolecules & Therapeutics
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• 제1권1호
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• pp.93-97
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• 1993

We have previously reported that an NADPH-dependent succinic semialdehyde reductase was purified homogeneously from bovine brain by several chromatographic procedures, and was found to be a monomeric protein with a molecular mass of 28 kDa (Cho et al., Eur. J. Biochem. 1993). Since succinic semialdehyde is an important intermediate in the ${\gamma}$-aminobutyrate(GABA) shunt and GABA level is associated with various forms of human neurological disorders, we have investigated the effects of anticonvulsant drugs on the succinic semialdehrde reductase. Among the drugs tested, sodium valproate and diphenylhydantoin inhibited the enzyme activity, while some other drugs, barbiturate and chlorpromazine, had no inhibitory effects on the enzyme activity. The purified enzyme was also injected as an immunogen into Balb/c mice to obtain monoclonal antibodies (mob) and several mobs to the protein were produced from the fusion experiments.

• Journal of Life Science
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• 제9권1호
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• pp.5-8
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• 1999

Helicobacter pylori is the etiologic agent of human gastritis and peptic ulceration and produces urease as the major protein component on its surface. H. pylori urease is known to serve as a major virulence factor and a potent immunogen. Deletion mutageneses were performed in the H. pylori urease accessory genes by using combinations of restriction enzymes and other DNA modifying enzymes in order to assess the function of these accessory gene products in the expression of the active urease. Selective disruptions in the accessory gene regions resulted in complete abolishment of the urease activity, which is consistent with other bacterial ureases. Interestingly, deletions in ureE-containing regions caused reduced expression of the structural enzyme subunits.

• 한국생활과학회지
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• 제2권1호
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• pp.17-24
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• 1993

Insulin (P)를 면역원으로 이용하여 insulin에 대한 균일한 항체집단의 대량유도 가능성 및 유도된 항체의 특성을 검토하였다. Insulin (P)에 의해 유도된 IgY는 insulin (M)에 의해 유도된 IgY 보다 조금 낮은 친화성을 나타내었으나 큰 차이는 없었다. 두 종류의 면역원에 의해 유도된 IgY는 대부분 insulin (M)을 인식하였으며, 같은 항원 특이성을 나타내었다.

• 한국동물위생학회지
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• 제33권2호
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• pp.173-176
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• 2010

Here, we report the suitability of using a resin-type chitosan formulae as a sustained-releasing form adjuvant in comparison with commercially well-known Freund's adjuvants. To induce the immunological responses, N-terminal region of Pasteurella multocida toxin was used as an antigen, which was found to be protective immunogen against P. multocida infection. Mice immunized with chitosan resin formulae showed statistically significant antibody induction (P<0.001) as much as that of Freund’s adjuvants. As a result, the resin-type sustained-releasing form chitosan formulae is thought to be a good candidate for a new type adjuvant.