• 한국식물병리학회지
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• 제3권2호
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• pp.108-113
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• 1987

효소결합항체법에 의한 벼검은줄오갈병 바이러스의 검정에서, 항원의 희석은 벼잎 $320\~2,450$배, 옥수수잎 $320\~5,120$배, 그리고 매개충인 애멸구는 $160\~2,560$배로 하는 것이 효과적이었다. 이상의 검정기준에 따른 성역별 월동약충(애멸구)의 보독충율 조사에서는 밀양 $3.0\%$, 칠곡 $2.3\%$ 그리고 선산은 $3.7\%$였다. 본 방법에 의하면 유묘접종법으로는 불가능한 채집과정에서 죽은 충에서도 바이러스 감염여부를 결정할 수 있었다. 포장에서 임의로 채집한 벼와 옥수수 각 100주씩을 공시하여 그중 외부병징이 없는 98주와 92주를 검정한 결과, 벼에서는 98주중 4주가 이병주로 나타났으며, 옥수수에서는 92주중 3주가 본 방법에 의해 감염된 것으로 나타나 포장에서의 잠복감염율은 벼에서는 $4.11\%$, 옥수수에서는 $3.25\%$로 나타났다. 따라서 효소결합항체법을 사용하면 병징이 잘 나타나지 않는 감염초기에도 검정이 가능하였다.

• 한국미생물·생명공학회지
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• 제13권3호
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• pp.265-271
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• 1985

Bacteriophage의 분류는 여러 가지 기준에 의해 수행될 수 있으나 본 연구에서는 L. casei bacterio-phage를 분류하기 위하여 3가지의 분류 기준이 사용되었다. 혈청학적 분류에서는 토끼에서 제조된 항혈청이 사용되었으며 항혈청에 의한 phage의 중화 효과는 대수적이었다. L. casei phage는 항혈청에 의한 중화률에 따라 3가지로 분류되었다. 여기서 Lac Y group은 새로운 종류임이 밝혀졌으며 Lac J 및 Lac S group은 지금까지의 보고와 동일한 결과를 보여주었다. 제한효소에 의한 phage DNA 절편의 비교에 의하여 Lac J group은 4종류의 subgroup으로 나누어졌으며, 숙주역의 차이에 의해서 Lac J-II group은 다시 3group으로 세분화되었다. 이러한 결과를 종합하면 L. casei S-1주의 bacte-riophage는 총 8종류로 나누어 질 수 있었다. 새로 분류된 Lac Y group의 YK phage는 직경이 95nm 정도의 icosahedral head와 길이 150nm, 넓이 20nm정도의 수축성 꼬리로 구성되어 있고 끝에 hexagonal baseplate가 있었다. YK phage DNA의 분자량은 약 86 Mdalton 이었다.

• Journal of Microbiology and Biotechnology
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• 제27권11호
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• pp.1971-1982
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• 2017

Piliated Lactobacillus rhamnosus (pLR) strains possess higher adherent capacity than non-piliated strains. The objective of this study was to isolate and characterize probiotic pLR strains in human fecal samples. To this end, mouse polyclonal antiserum (anti-SpaA) against the recombinant pilus protein (SpaA) of L. rhamnosus strain GG (LGG) was prepared and tested for its reactivity and specificity. With the anti-SpaA, a method combining the de Man, Rogosa, and Sharpe (MRS) agar plating separation and colony immunoblotting (CIB) was developed to isolate pLR from 124 human fecal samples. The genetic and phenotypic characteristics of the resultant pLR isolates were compared by randomly amplified polymorphic DNA (RAPD) fingerprinting, and examination of adhesion to Caco-2 cells, hydrophobicity, autoaggregation, and in vitro gastrointestinal tolerance. Anti-SpaA specifically reacted with three pLR strains of 25 test strains, as assessed by western blotting, immunofluorescence flow cytometry, and immunoelectron microscopy (IEM) assays. The optimized MRS agar separation plus anti-SpaA-based CIB procedure could quantitatively detect $2.5{\times}10^3CFU/ml$ of pLR colonies spiked in $10^6CFU/ml$ of background bacteria. Eight pLR strains were identified in 124 human fecal samples, and were confirmed by 16S RNA gene sequencing and IEM identification. RAPD fingerprinting of the pLR strains revealed seven different patterns, of which only two isolates from infants showed the same RAPD profiles with LGG. Strain PLR06 was obtained with high adhesion and autoaggregation activities, hydrophobicity, and gastrointestinal tolerance. Anti-SpaA-based CIB is a rapid and inexpensive method for the preliminary screening of novel adherent L. rhamnosus strains for commercial purposes.

• 한국응용곤충학회지
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• 제23권1호
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• pp.15-21
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• 1984

강낭콩에서 종자전염된 바이러스병을 동정하기 위하여 각지역 농가 및 시장에서 종자를 수집파종하여 종자전염을 조사한 결과 수원, 장수, 진주에서 채집한 종자가 $2.0\~3.5\%$ 이병되어 있었으며 종자전염된 바이러스를 지표식물에 접종한 결과 강낭콩에 상엽에 모자익, Chenopodium quinoa의 접종엽에 대형의 국부병반이 형성되었다. Dip법에 의하여 시료를 제작하여 입자를 관찰한 결과 약 750nm의 계상입자가 관찰되었다. 이병 강낭콩의 조직에서 Cylinder와 Pinwheel형 및 bundle형의 봉입체(Inclusion hody)가 관찰되었다. 복숭아진딧물에 의하여 충매전염 되었으며 즙액전염된 이병주로부터 채집한 종자의 전염율은 $18.2\%$였다. 물리적성질은 내희석성이 $1,000\~5,000$배 였으며 내보존성은 실온에서 $2\~3$일, 내열성은 $55\~60^{\circ}C$였다. 항혈청검정 결과 혼합침강반응에서 양성의 반응을 나타냈으며 SDS처리에 의한 한천내이중확산법에서 BCMV 항혈청과 band를 형성하였으며 AzMV와는 spur가 형성되지 않았으며 CAMV와는 spur를 형성하였다. 이상의 결과로 강낭콩에서 종자전염된 바이러스는 Bean common mosaic virus로 동정되었다.

• Journal of Microbiology and Biotechnology
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• 제16권10호
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• pp.1529-1536
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• 2006

We cloned a gene of ompH(D:4) from pigs infected with P. multocida D:4 in Korea [16]. The gene is composed of 1,026 nucleotides coding 342 amino acids (aa) with a signal peptide of 20 aa (GenBank accession number AY603962). In this study, we analyzed the ability of the ompH(D:4) to induce protective immunity against a wild-type challenge in mice. To determine appropriate epitope(s) of the gene, one full and three different types of truncated genes of the ompH(D:4) were constructed by PCR using pET32a or pRSET B as vectors. They were named ompH(D:4)-F (1,026 bp [1-1026] encoding 342 aa), ompH(D:4)-t1 (693 bp [55-747] encoding 231 aa), ompH(D:4)-t2 (561 bp [187-747] encoding 187 aa), and ompH(D:4)-t3 (540 bp [487-1026] encoding 180 aa), respectively. The genes were successfully expressed in Escherichia coli BL21(DE3). Their gene products, polypeptides, OmpH(D:4)-F, -t1, -t2, and -t3, were purified individually using nickel-nitrilotriacetic acid (Ni-NTA) affinity column chromatography. Their $M_rs$ were determined to be 54.6, 29, 24, and 23.2 kDa, respectively, using SDS-PAGE. Antisera against the four kinds of polypeptides were generated in mice for protective immunity analyses. Some $50{\mu}g$ of the four kinds of polypeptides were individually provided intraperitoneally with mice (n=20) as immunogens. The titer of post-immunized antiserum revealed that it grew remarkably compared with pre-antiserum. The lethal dose of the wild-type pathogen was determined at $10{\mu}l$ of live P. multocida D:4 through direct intraperitoneal (IP) injection, into post-immune mice (n=5, three times). Some thirty days later, the lethal dose ($10{\mu}l$) of live pathogen was challenged into the immunized mouse groups [OmpH(D:4)-F, -t1, -t2, and -t3; n=20 each, two times] as well as positive and negative control groups. As compared within samples, the OmpH(D:4)-F-immunized groups showed lower immune ability than the OmpH(D:4)-t1, -t2, and -t3. The results show that the truncated-OmpH(D:4)-t1, -t2, and -t3 can be used for an effective vaccine candidate against swine atrophic rhinitis caused by pathogenic P. multocida (D:4) isolated in Korea.

• Animal cells and systems
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• 제2권1호
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• pp.139-144
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• 1998

I examined the projection of glutamatergic neurons in the lateral reticular nucleus into ansiform (crus l and ll) and paramedian lobules in the rat cerebellum using immunocytochemical methods with antiserum against glutamate combined with WGA-HRP histochemistry. The projections of glutamatergic neurons from the lateral reticular nucleus to crus l were most extensive in number among the three injection cases and the majority of projections originated at the dorsal to dorsomedial region of the ipsilateral magnocellular nucleus. Glutamate-immunoreactive cells projecting to crus ll were less extensive in number than those projecting to crus l and were mainly localized at the dorsomedial portion of the ipsilateral magnocellular nucleus. Double-labelled neurons projecting to crus l or crux ll were also located at ipsilateral subtrigeminal as well as contralateral magnocellular nuclei. Glutamatergic neurons projecting to paramedian lobules were moderate in number and mainly located at the dorsal area of the ipsilateral magnocellular nucleus. A few double-labelled cells were also found at ipsilateral subtrigeminal or contralateral magnocellular nuclei. The present study suggests that glutamate-immunoreactive neurons at the dorsal to dorsomedial magnocellular division of the lateral reticular nucleus may participate in the excitatory control of target neuronal activities at ipsilateral, posterior hemispheric lobules of the rat cerebellum.

• International Journal of Industrial Entomology and Biomaterials
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• 제1권2호
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• pp.131-135
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• 2000

The vitellin of the fireflies, Luciola unmunsana and L. lateralis was characterized. The vitellin of L. unmunfon is composed of two subunits, designated Vnl (195 kDa) and Vn2 (185 kDa) in SDS-polyacryamide gel electrophoresis. These two subunits of vitellin of L. unmunsana gradually decreased during embryogenesis. As expected, these protein bands were presented in female adult hemolymph and egg extracts, but not in male. The vitellin of L. lateralis is also composed of two subunits, designated Vnl (195 kDa) and Vn2 (180 kDa) in SDS-PAGEi and these two protein bands gradually decreased during embryogenesis. Western blot analysis using each of polyclonal antiserum against vitellins of L. unmunsana and L. lateralis showed that two antisera strongly crossereacted with vitellin subunits of L. unmunsana and L. lateralis, suggesting that vitellins of L. unmunsana and L. lateralis have similarity with each other.

• 대한수의학회지
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• 제31권2호
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• pp.223-228
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• 1991

Most progesterone enzyme immunoassays(EIA) are used liquid-phase double-antibody or single-antibody seperation. These methods consume considerable time and reagents because of the requirements for several washing and centrifugation steps involving the reactants. Because of these several problems, we were prompted to develop an effective enzyme-linked immunosorbent assay(ELISA) system that would be equal or superior to RIA for assay of progesterone. The results were obtained as follows. 1. Cross reaction of the progesterone antiserum with other steroids determined was shown with progesterone(100%), $11{\alpha}$-deoxycorti-costerone(2.271%), but the other steroids were shown below 0.9%. 2. Standard curve for progesterone ELISA was shown available difference according to progesterone concentration from 0 to 1,000pg/ml. 3. The lower limit of sensitivity was 0.2pg/well 4. Progesterone concentration was 1.6ng/ml for before parturition, and that was below 0.5ng/ml for after parturition. This development enzyme-linked immunosorbent assay for progesterone can be detected pregnancy diagnosis in cattle, and also applicable 10 research on physiological function including such as reproductive disorders.

• 대한수의학회지
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• 제5권1호
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• pp.43-57
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• 1965

It has been-reported that rabbit serum exhibit an inhibitory action on avian infectious bronchitis virus in embryonating chicken embryo. In this thesis, the biological, serological, physical and chemical properties of normal rabbit serum on the effect of the virus propagation were studied. Throughout the studies, the following experimental results 'were obtained and summarized here. 1. An inhibitory action of rabbit serum on avian infectious bronchitis vrius is due to the normal serum constituents. 2. The nature of the neutralization between normal rabbit serum and the virus is similar to that of the specific antiserum and the virus. 3. Rabbit serum, heat inactivated at $56^{\circ}C$, for 30 minutes, showed its average $log_{10}El,D_{50}Nl$ of 3.7. 4. The inhibitory compound present in the normal rabbit serum is inactivated by means of 5 per cent trypsin, 0.01 M potassium periodate, and absorbed to zymosan. 5. The inhibitory compound was not affected by 0.05 M trichloroacetic acid and 0.005M $KH_2PO_4$. 6. The higher the temperature of heat inactivation of rabbit serum caused the lesser the neutralizing effect on the virus. Heating the serum at $66^{\circ}C$, for 30 minutes brought about a complete loss of the neutralizing index of the serum. 7. No ions, as a cofactor, was incorporated to the inhibitory action of rabbit serum on the virus. 8. The inhibitory compound amays be found in a fraction of serum globulin.

• 한국식품위생안전성학회지
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• 제9권3호
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• pp.141-149
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• 1994

To investigate the distribution of Vibrio parahaemopyticus in the southern 4 coastal areas, Vibrio parahaemolyticus was isolated from seawater, shellfish and sediment from May to October in 1991, and antimicrobial effect of grapefruit seed extract (GFSE) on the growth of isolated strains were examined. In the 120 sample from 4 coastal areas, 16 strains of Vibrio parahaemolyticus were isolated and identified. The distribution serotype of isolated strains was 10 types of monovalent k-antiserum. Especially k-5 and k-28 were highly distribyted with 3 and 4 strains. 31.3% of isolated strains showed positive on Kanagawa phenomenon test. All isolated Vibrio parahaemolyticus were sensitive to chloramphenicol and gentamycin, 5 and 6 strains were resistant to streptomycin and colistin, respectively. Isolated strains were compared with geographical, month and sample. The distribution of 16 isolated Vibrio parahamolyticus was high at Hadong with 50%(8 strains), on July with 43.8%(7 strains) and from seawater with 37.5%(6 strains) respectively. Minimal inhibitory level of GFSE to Vibrio parahaemolyticus was 50 ppm. With 100 ppm treatment of GFSE, the destroy of cell membrane function, outflow of cell ingredients and ghost morphology of cell were investigated.