• 대한수의학회지
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• 제33권3호
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• pp.443-454
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• 1993

Shiga-like toxin-II(SLT-II)-producing Escherichia coli 0157 : H7 strain B2387이 분비하는 SLT-II가 gnotobiotic자돈에서의 뇌혈관 병변을 일으키는 pathogenesis에 관해서 실험을 했다. 제왕절개 수술로 태어난 자돈들을 두 그룹으로 나누어서, 한 그룹에는 SLT-II 중화항체를 포함한 혈청을 구강을 통해서 수동면역을 시키고, 또다른 한 그룹에는 SLT-II 중화항체가 포함되어 있지 않은 혈청을 구강을 통해서 수동면역시켰다. 24시간후 두 그룹 모두에게 SLT-II producing Escherichia coli O157 : H7 strain B2387를 구강으로 접종했다. SLT-II 중화항체가 포함되어 있지 않은 혈청으로 수동면역시킨 그룹의 자돈들은 설사와 맹결장염, 신경증상, 뇌혈관병변을 일으키고, plasma의 prostacyclin의 level이 증가했다. 하지만 SLT-II 중화항체가 포함되어 있는 혈청으로 수동면역시킨 그룹의 자돈들은 설사와 맹결장염은 유발했지만, 신경증상과 뇌혈관병변은 관찰되지 않았고, prostacyclin의 level도 증가하지 않았다. 이런 실험결과는 SLT-II 중화항체는 뇌혈관병변은 방어하지만 맹결장염은 방어하지 못한다는 의미를 나타내며, prostacylin의 증가는 뇌혈관의 endothelium의 병변을 의미한다.

• Journal of Microbiology
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• 제34권2호
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• pp.151-157
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• 1996

By PCR amplification using the sequence of the previously cloned shiga-like toxin II DNA, a gene encoding it has been cloned from an isolate of healthy Korean native bovine feces, Escherichia coli KSC109. The nucleotide sequence s included tow open reading frames coding for 319 and 89 amino acids corresponding to A and B subunits, respectively. Comparison of the nucleotide and predicted amino acid sequences of newly cloned gene (slt-II) with those of others in the SLT-II family revealed completely identical homology with SLT-II cloned previously from bacteriophabe DNA of E. coli 933 derived from a patient with hemorrhagic colities. In addition, the sequence homology of SLT-II with SLT-II variant form bovine was more than 95% at both the nucleotide and protein levels. Overexpression of SLT-II recombinant gene by induction with IPTG using an E, coli hostvector, system was conducted and the correctly processed products with active mature form exhibited 1000-fold higher cytotoxycity for Vero cells than that form original strain.

• Journal of Microbiology
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• 제37권3호
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• pp.168-174
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• 1999

Escherichia coli O157 is an important pathogenic organism which causes diarrhea, haemorrhagic colitis, and haemolytic ureamic syndrome (HUS) in human. E. coli O157 KNIH317 was isolated form patients suffering with HUS in Korea. We designed a primer set for cloning shiga-like toxin (slt) gene. The amplified PCR product was used to Southern and colony hybridization as a probe. As a result, we cloned 4.5-kb KpnI fragment containing the slt gene encoding shiga-like toxin from chromosomal DNA of E. coli O157 KNIH317. This recombinant plasmid was named pOVT45. E. coli XL1-Blue harboring pOVT45 showed cytotoxicity in Vero cells. We sequenced the slt gene of this strain. The A-subunit gene of the slt was composed of 960 base pairs with ATG initiation codon and TAA terminationcodon. The B-subunit was composed of 270 base paris with ATG initiation codon and TGA termination codon. Nucleotide sequence comparison of the slt gene exhibited 100%, 98.4%, 93.7%, and 93.7% identity with that of shiga-like toxin type II (sltII) of E. coli bacteriophage 933W, variant slt of E. coli, slt of E. coli, and variant sltII of E. coli, respectively. From these results, it was concluded that the cloned slt gene belongs to SltII family and that the strain used in this study may be a lysogeny of E. coli bcteriphage 933W.

• 대한수의학회지
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• 제38권1호
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• pp.173-181
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• 1998

Esherichia coli O157 : H7의 slt I, slt II, uid A 및 eaeA 4종 유전자를 동시에 검출하기 위한 multiplex PCR 기법을 확립하고 쇠고기중 직접 E coli O157 : H7 검출시험을 실시하였다. 4 set의 primers를 이용한 multiplex PCR 기법으로 31종의 장내세균에 대한 특이성을 조사한 결과 E coli O157 : H7 에서 1,087bp (eae A), 584bp (slt II), 348bp (slt I) 또는 252bp (uid A)크기의 DNA를 동시에 특이적으로 검출할 수 있었다. E coli O157 : H7 15주는 모두 uid A 및 eae A 유전자가 동시에 검출되었고, 다른 장내세균에서는 검출되지 않았다. slt I 또는 slt II 유전자를 가지고 있는 E coli 표준균주 24종을 이용하여 multiplex PCR 기법과 Vero cell cytotoxicity assay을 비교검사한 결과 베로톡신 산생능과 PCR법의 결과는 일치하였다. mutiplex PCR 기법의 쇠고기중 검출한계는 modified EC(mEC)에서 증균없이는 E coli O157 : H7균 $10^4cells/g$ 이상에서 검출이 가능하였으나 mEC에 1차 증균후 modified TSB 증균하였을 경우에는 10cells/g이하까지도 검출이 가능하였다. 개발된 multiplex PCR 기법을 쇠고기 40종에 직접 적용한 결과 E coli O157 : H7은 검출되지 않았으나 slt I 및 slt II유전자를 가지고 있는 E coli 4종이 검출되었으며, 이들의 혈청형은 O6, O112, O115 및 O139 였다. 이 연구에서 개발된 multiplex PCR은 쇠고기중 E coli O157 : H7을 신속하고 특이적으로 검출하는데 사용할 수 있을 것으로 사료된다.

• 대한의생명과학회지
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• 제4권1호
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• pp.43-56
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• 1998

최근 전세계적으로 문제가 되고 있는 장출형성 대장균 O157:H7을 분리배양 및 동정 없이 바로 시료를 분석하여 신속하게 검출하기 위한 다중 중합효소 연쇄반응 (multiplex PCR) 기법을 확립하고, 이 기법을 이용하여 국내 분리 균주 중에서 SLT-I.II, eaeA, 60-MDa plasmid gene을 가지고 있는 대장균을 유전자 수준에서 검출하고자 하였다. 장출혈성 대장균 O157:H7이 가진 SLT-I.II, 60-MDa plasmid 유전자들에 대한 특이 oligonucleotide primers (MK1'-MK2', NAE19-NAE20, MFSIF-MFSIR)를 함께 동시에 반응 완충액에 넣어 다중 중합효소 연쇄반응을 시행한 결과 317bp (eaeA), 228bp (SLT-I.II), 167bp (60-MDa plasmid)의 PCR 증폭 DNA생성물을 표준균주 (E. coli ATCC 35150)에서는 확인할 수 있었지만, 기타 다른 병원성 장내세균 13세균 13균주에서는 band를 확인할 수 없었다. 한편 다중 중합효소 연쇄반응의 template DNA 추출 방법에 따른 PCR 결과를 비교하였다. 각각의 DNA 추출 방법 중 boiling lysis 방법이 신속하고 간편하여 장출혈성 대장균 O157:H7에 의한 식중독의 임상진단에 다중 중합효소 연쇄반응 (multiplex PCR) 적용하는 데에는 boiling lysis법을 이용하는 것이 가장 적합한 방법으로 확인되었다.

• 대한수의학회지
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• 제39권2호
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• pp.338-342
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• 1999

Twenty three strains of Escherichia (E) coli O157 : H7 isolated from Korea, Japan, USA were analyzed by pulsed-field gel electrophoresis (PFGE) of XbaI-digested chromosomal DNA and multiplex polymerase chain reaction. Various PFGE patterns of E. coli O157 : H7 were found on the same farm. Most of the E, coli O157 : H7 strains had shiga-like toxin (slt) II gene only (43.5%) or both slt I and slt II genes(30.4%). eaeA gene was highly conserved in the E. coli O157 : H7. There was no correlation between PFGE and slt gene patterns. The results indicate that various genotypes of E. coli O157 : H7 have spread throughout the country and genomic DNA patterns generated by PFGE are highly specific for different strains and have significant value in epidemiologic investigations of infectious disease outbreaks.

• Weed & Turfgrass Science
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• 제1권4호
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• pp.57-63
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• 2012

본 연구는 무작위 분자마커(RAPD)를 이용한 우리나라 골프장에서 이용되고 있는 잔디 13개 잔디품종의 유전적 다형성을 조사하여 보다 효과적인 골프장 관리를 위한 유전적 정보를 제공하고자 조사하였다. 본 연구에서 사용한 54개의 random hexamer primer를 이용하여 RAPD분석을 실시한 결과 13~54개의 다형성 밴드를 형성하였으며 primer당 평균 30.7개의 다형성 밴드를 확인할 수 있었다. RAPD분석 결과 13개의 잔디품종은 크게 3개의 그룹으로 나눌 수 있었다. Group I은 Shadow II, Aurora Gold, Little Big Horn Blue, PennA-1, PennA-4, Group II는 Midnight II, Prosperity, Moonlight SLT, Bright Star SLT, Silver Dollar, Group III은 Olympic Gold Turf-Type, Silver Star Turf-Type, Tar Heel II Turf-Type을 포함하였다. 13개 잔디 품종의 유전적 근연 정도는 0.039~1.0으로 나타났으며, Group III이 유전적 근연 정도가 가장 높게 나타났다. 이상의 결과를 통해 향후, 잔디 종 또는 이종간의 유전적 다양성의 상호관계나 차이점을 규명하기 위해서는 형태적인 특성과 SCARs 마커와 같은 특이적인 분자마커에 대한 연구가 추가적으로 필요할 것으로 사료된다.

• 동의생리병리학회지
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• 제21권5호
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• pp.1278-1284
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• 2007

Solanum Iyratum Thunb (Solanaceae) has multiple applications in korean traditional medicine because of its cytotoxic, immunological and anti-inflammatory activities. However, no study on the anti-arthritic activity of Solanum Iyratum Thunb has been reported in vivo. Rheumatoid arthritis (RA) is a systemic autoimmune disease with chronic inflammation characterized by hyperplasia of synovial cells in affected joints, which ultimately leads to the destruction of cartilage and bone. Cytokine production were assessed during CIA(collagen-induced arthritis) model mice in lymph node (LN), in knee joint and spleen, using ELISA. DBA/1j mice were immunized with bovine type II collagen. After a second collagen immunization, mice were treated with Solanum Iyratum Thunb (SLT) orally at 400, 200 mg/kg once a day for 4 weeks. The severity of arthritis within the knee joints was evaluated by histological assessment of cartilage destruction and pannus formation. SLT significantly suppressed the progression of CIA and inhibited the production of TNF-alpha and IFN-gamma in serum and spleen cell culture supernatant. The erosion of cartilage was dramatically reduced in mouse knees after treatment with SLT. In conclusion, our results demonstrates that SLT significantly suppressed the progression of CIA. This action was characterized by suppression of arthritis index, cartilage erosion and synovial cell infiltration and the decreased production of $TNF-{\alpha}$, $IFN-{\gamma}$, CD4+, CD19+, CD3+/CD69+ cells (in lymph node), CD11b+/Gr-1 + (in knee joint).

• Mycobiology
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• 제45권4호
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• pp.362-369
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• 2017

We assessed the regulation of cryparin, a class II hydrophobin, using three representative mitogen-activated protein kinase (MAPK) pathways in Cryphonectria parasitica. Mutation of the CpSlt2 gene, an ortholog of yeast SLT2 in the cell wall integrity (CWI) pathway, resulted in a dramatic decrease in cryparin production. Similarly, a mutant of the CpBck1 gene, a MAP kinase kinase kinase gene in the CWI pathway, showed decreased cryparin production. Additionally, mutation of the cpmk1 gene, an ortholog of yeast HOG1, showed decreased cryparin production. However, mutation of the cpmk2 gene, an ortholog of yeast Kss1/Fus3, showed increased cryparin production. The easy-wet phenotype and accumulation of the cryparin transcript in corresponding mutants were consistent with the cryparin production results. In silico analysis of the promoter region of the cryparin gene revealed the presence of binding motifs related to downstream transcription factors of CWI, HOG1, and pheromone responsive pathways including MADS-box- and Ste12-binding domains. Real-time reverse transcriptase PCR analyses indicated that both CpRlm1, an ortholog of yeast RLM1 in the CWI pathway, and cpst12, an ortholog of yeast STE12 in the mating pathway, showed significantly reduced transcription levels in the mutant strains showing lower cryparin production in C. prasitica. However, the transcription of CpMcm1, an ortholog of yeast MCM1, did not correlate with that of the mutant strains showing downregulation of cryparin. These results indicate that three representative MAPK pathways played a role in regulating cryparin production. However, regulation varied depending on the MAPK pathways: the CWI and HOG1 pathways were stimulatory, whereas the pheromone-responsive MAPK was repressive.

• 한국환경복원기술학회지
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• 제12권3호
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• pp.72-82
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• 2009

Research was initiated to investigate early establislment characteristics and germination pattern of perennial ryegrass (PR, Lolium perenne L.). Eight varieties of PR were evaluated in the study. An alternative environmental condition for a PR germination test required by International Seed Testing Association (ISTA) was applied in the experiment, consisting of 8-hr light at $25^{\circ}C$ and 16-hr dark at $15^{\circ}C$ (ISTA conditions). Daily and cumulative germination pattern were measured and analyzed daily for 30 days. Significant differences were observed in germination pattern, days to the first germination, days to 50% germination, days to 75% germination, and final germination rate. Germination rates were variable among the varieties, being 66.00 to 93.75% at the end of study. There were considerable variations in early germination characteristics and germination pattern among PR varieties. The first germination in all entries tested was initiated from 3 to 5 DAS (days after seeding) under ISTA conditions, being 2 days in differences among the varieties. It was 4 to 7 DAS in days to the 50% germination, being 1 to 3 days later from the first germination and three days were variable among them. Days to the 75% germination were 5.93 to 10.50 DAS under ISTA conditions, being 4.57 days in variety differences. Pennant II and Brightstar II were the fastest varieties. The slowest ones were Revenge GLX and Brightstar SLT. Considering days to the first germination, days to 75% germination, and germination pattern etc, Pennant II and Brightstar II were regarded as excellent varieties under ISTA conditions. From tills study, information on differences in germination characteristics and pattern would be usefully applied for golf course design and construction, when established with PR.