• 생명과학회지
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• 제28권9호
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• pp.1118-1126
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• 2018

본 논문은 양식 어류에게 많은 경제적 피해를 입히고 있는 다양한 어류질병세균 중에서 연쇄구균증에 대한 일반적인 특성과 종류, 진단 방법에 대하여 기존에 연구 보고된 논문을 기반으로 알아보고자 한다. 대표적인 어류연쇄구균증의 원인체로는 Lactococcus garvieae, Streptococcus iniae, S. parauberis가 있다. 연쇄구균증에 감염 시 나타나는 증상으로는 체색변화, 안구이상, 아가미 퇴색, 출혈, 복부팽만, 신장과 비장의 종대 등의 보이다 폐사가 일어난다. 또한 수온이 상승하는 6월부터 10월까지 주로 일어나며 집단적으로 발병하여 폐사가 일어난다. 현재 연쇄구균증을 진단하기 위한 기술로는 16S rRNA, 16S-23S rRNA intergenic spacer region (ISR), Random Amplified polymorphic DNA (RAPD), Ribotyion (RT), Loop-mediated isothermal amplification (LAMP) 등이 있다. 이중에서 현장에서 적용 가능성이 높은 LAMP법이 각광을 받고 있지만 결과 확인 등의 문제로 인하여 현재는 어려움을 겪고 있는 실정이다. 이에 현장에서 진단부터 결과 확인까지 손쉽게 사용할 수 있도록 문제점을 보완하면 연쇄구균증에 대한 경제적 손실을 최소화 할 것으로 사료된다.

• 대한의생명과학회지
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• 제11권4호
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• pp.455-463
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• 2005

Resistance to isoniazid (INH), which is one of the most important drugs in Mycobacterium tuberculosis chemotherapy, has been associated with mutations in genes encoding the mycobacterial catalse-peroxidase (katG), the enoyl acyl carrier protein (ACP) reductase (inhA), alkyl hydroperoxide reductase (ahpC), beta-ketoacyl acyl carrier protein synthase (kasA), and NADH dehydrogenase (ndh). In this study, we examined INH-resistance related genes in 50 INH-resistant and 24 INH-susceptible isolates by PCR-sequence analysis. In brief, mutations at the katG gene were found at codon 315 alone (2/50), at codon 463 alone (19/50), and both at 315 and 463 (29/50). However, while mutations at codon 315 were only detected in INH-resistant isolates, mutations at codon 463 were also detected in INH-susceptible isolates indicating mutations at 463 alone do not seem to confer resistance to INH. Similar to the case of katG 463, some of inhA mutations were also found among INH-susceptible isolates. For example, whereas mutations at 8 upstream of the start codon (UPS) and 15 UPS of the inhA gene were detected only in INH-resistant isolates, mutations at 101, 115, and 125 UPS were detected only in INH-susceptible isolates. Many different kinds of mutations were detected in INH­resistant isolates at ahpC, oxyR gene, and intergenic region of the oxyR-ahpC genes. Howerver, the mutations were not found oxyR and the intergenic regions in INH-susceptible isolates. No mutations were found at either kasA or at ndh gene among INH-resistant isolates. In conclusion, some of mutations such as katG 315, inhA promotor region, and oxyR-ahpC seem to be strongly related to INH-resistance. Currently we are developing a molecular diagnostic method based on these results.

• 한국미생물·생명공학회지
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• 제30권4호
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• pp.312-319
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• 2002

저온성 균인 Sporosarcina psychrophilia의 염색체 DNA를 추출하여 Sau3AI으로 부분 절단하고 pUC19 vector에 ligation 시킨 후, Escherichia coli pyrB mutant 균주에 형질전환하여 uracil이 없는 AB배지에서 생존하는 균주를 선택한 후, 그 plasmid를 분리하여 pSMI과 pSM2라고 명명하였다. 두 plasmid의 염기배열을 결정한 결과 pSM2 insert DNA는 pSMl insert DNA 부분을 포함하는 2,606 nucleotide 단편이었다. 염기서열을 분석하였을 때 이것은 1개의 완전한 open reading frame(ORF)과 2개의 부분 ORFs를 포함하고 있었다. 두 번째 위치한 완전한 ORF는 Bacillus caldolyticus aspartate transcarbamylase(pyrB)와 아미노산 서열 수준에서 59% 상동성을 보였고, 첫 번째와 세 번째 위치한 부분적 ORFs는 각각 Bacillus속의 uracil permease(pyrP)와 dihydoorotase(pyrC)와 높은 상동성을 보였다. 그리고 pyrB와 pyrP사이에 intergenic 부분에는 잠재적인 terminator, antiterminator, anti-antiterminator 구조를 포함하고 있었다. 이러한 결과는 S. psychrophilia pyrimidine 생합성에 관련된 유전자들은 다른 Bacillus속에서 알려진 바와 같이 유전자군을 형성하고 있을 것으로 추정했다. S. psychrophilia pyrB 유전자의 생성물을 과다발현 시키고 정제해서 그 단백질을 SDS-PAGE로 확인한 결과 27 kDa 부근에서 band를 확인할 수 있었으며, 정제한 단백질도 ATCase 효소활성을 지니고 있었다.

• 식물병연구
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• 제26권2호
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• pp.79-87
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• 2020

Fusarium oxysporum f. sp. fragariae (Fof) 에 의한 딸기 시들음병은 국내 딸기재배에서 가장 중요한 병해 중 하나이다. 국내 발생하는 Fof의 특성을 분석하고자 시들음병균의 유전적 다양성, 병원성과 살균제 반응을 조사하였다. 분리균은 Fo080701를 제외한 모든 균주에서 Fof 특이적 primer에 증폭되었다. 분리균의 nuclear ribosomal intergenic spacer region과 EF-1α sequences 분석 결과 3개의 lineage를 형성하였다. 대부분의 분리균은 lineage 1에 속하였으며 lineage 3에 3개 균주와 lineage 2에 1개 균주가 포함되었다. 분리된 모든 균주는 설향품종에 병원성을 보였다. Prochloraz는 DNA lineage 2에 속하는 Fo080701균주를 제외하곤 시들음병균의 EC₅₀값이 0.02-0.1 ㎍/ml로 낮은 농도에서 효과적으로 균사 생장을 억제하였다. Metconazole의 EC₅₀값도 0.04-0.22 ㎍/ml로 prochloraz와 비슷한 억제 효과를 보였다. Pyraclostrobin의 EC₅₀값은 0.23-168.01 ㎍/ml로 균주에 따라 차이가 컸다. 딸기 재배포장에서 boscalid+fludioxonil, fluxapyroxad+pyraclostrobin, prochloraz manganese이 딸기 시들음병 방제에 효과적이었다.

• 한국어병학회지
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• 제19권2호
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• pp.127-139
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• 2006

2002년에서 2004년에 걸쳐, 우리 나라의 넙치 양식장에서 질병의 증상이 보이는 중간 육성 단계의 넙치를 대상으로 다수의 Vibrio속 세균을 분리 동정하였으며 그 중에서 V. ichthyoenteri균이 높은 비율을 차지하고 있었다. 일반적으로 V. ichthyoenteri는 넙치의 자어기 장관백탁증의 원인균으로 잘 알려져 있으며, 미성어의 병어로부터 분리되었다는 점에서 여러 크기의 넙치 및 종묘 생산에 미치는 영향을 알고자 하였다. 본 연구는 미성어 넙치에서 분리된 균주그룹과 장관백탁증에 걸린 넙치 자어에서 분리한 V. ichthyoenteri 참조 균주 두 그룹간의 생화학적 및 생리학적 성상, 유전학적 특성을 비교 분석함으로서 분리 균주를 V. ichthyoenteri로 동정하고, 이들의 어병학적 특성을 조사하였다.실험 결과 시험 균주와 참조 균주간의 생화학적 특성과 생리학적 성상이 거의 동일한 것으로 나타났다.본 연구에서 분리 균주와 참조 균주의 16S-23S rRNA intergenic space (IGS) region을 cloning하여 염기 서열을 분석한결과 3개의 참조 균주와 19개의 분리 균주들은 V. ichthyoenteri 참조 균주와 99.1 ~ 100% 동일하였으며 V. ichthyoenteri는 3개 이상의 PCR products를 보였다. 이들 각각에 대한 tRNA type을 분석한 결과, no-tRNA, tRNAGlu(TTC) type 그리고 tRNAIle(GAT)와 tRNAAla(TGC) 를 coding하는 3 가지 tRNA type을 밝혔다.

• The Plant Pathology Journal
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• 제33권2호
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• pp.140-147
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• 2017

Fusarium wilts of strawberry, caused by Fusarium oxysporum f. sp. fragariae, is a serious soil-borne disease. Fusarium wilt causes dramatic yield losses in commercial strawberry production and it is a very stubborn disease to control. Reliable chemical control of strawberry Fusarium wilt disease is not yet available. Moreover, other well-known F. oxysporum have different genetic information from F. oxysporum f. sp. fragariae. This analysis investigates the genetic diversity of strawberry Fusairum wilt pathogen. In total, 110 pathogens were isolated from three major strawberry production regions, namely Sukok, Hadong, Sancheong in Gyeongnam province in South Korea. The isolates were confirmed using F. oxysporum f. sp. fragariae species-specific primer sets. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analyses were executed using the internal transcribed spacer, intergenic spacer, translation elongation factor1-${\alpha}$, and ${\beta}$-tubulin genes of the pathogens and four restriction enzymes: AluI, HhaI, HinP1I and HpyCH4V. Regarding results, there were diverse patterns in the three gene regions except for the ${\beta}$-tubulin gene region. Correlation analysis of strawberry cultivation region, cultivation method, variety, and phenotype of isolated pathogen, confirmed that genetic diversity depended on the classification of the cultivated region.

• 한국버섯학회지
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• 제12권3호
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• pp.145-153
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• 2014

Pleurotus has increased rapidly production and consumption because of highly nutritional value, natural healthy food and so on. The basic studies for Pleurotus need for development of mushroom industry. This study was to investigate the cultural characteristics among 15 strains of 6 species and to analyze their phylogenetic relationships. The cultural characteristics were investigated by mycelial growth activity at different media, temperature and pH. The optimum media for mycelial growth were YM and MCM in most species. The optimum temperature for mycelial growth were $25^{\circ}C$ and $30^{\circ}C$. The optimum pH for mycelial growth were widly range from pH 5.1 to 7.4. Through the RFLP (restriction fragment length polymorphism) of IGS (intergenic spacer) I region in ribosomal DNA, it was analyzed phylogeny of interspecies and intraspecies. Each species was discriminated well as isolates within each species formed clade to be distinguished other species. P. florida was highly similar to P. floridanus, and P. flabellatus was P. cornucopiae. P. fuscus var. ferulae was highly similar to P. eryngii but discriminated different species in analysis of RFLP of IGS I region and showed different characteristics in mycelial culture. RFLP of IGS I region was useful of studying phylogenetic relationships of species and population.

• 대한수의학회지
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• 제35권1호
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• pp.95-105
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• 1995

The human homolog of position specific element of mouse Hoxa-7 was studied using transgene. It contains a 1.1 kb human DNA (HCR)- a homolog to the intergenic region between Hoxa-7 and -9, which directs the position specific expression of Hoxa-7-, tk promoter, LacZ (${\beta}$-galactosidase) gene as a reporter, and polyadenylation signal of SV40 large T antigen. It was injected into the mice embryos, and the resulting transgenic embryos were analysed through PCR as well as genomic Southern blotting with placenta DNA. Out of 20 embryos analysed, two were transgenic. Among them, one transgenic embryo expressed transgene when stained with X-gal. The expression pattern was in analogy to that of the mouse Hoxa-7, showing spatially restricted expression pattern, Since the expression of ${\beta}$-galactosidase is regulated by the upstream human HCR sequence, it implies that the HCR is the plausible position specific regulatory element of human.

• Journal of Crop Science and Biotechnology
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• 제11권4호
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• pp.237-242
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• 2008

A liguleless mutant, which showed complete loss of lamina joint region at the junction between leaf blade and leaf sheath, was isolated from a Ds insertional mutants derived from the source cultivar, Dongjin. This mutant could not affect other developmental patterns like phyllotaxis. Southern blot analysis, using GUS as a probe, revealed that the liguleless mutant contained three Ds copies transposed in the rice genome. Among the four genomic sequences flanking the Ds, one was mapped in the intergenic region (31661640 - 31661759), and the other two predicted a protein kinase domain (12098980 - 12098667) as an original insertion site within a starter line used for massive production of Ds insertional mutant lines. Another predicted and inserted in first exon of liguleless 1 protein (OsLG1) that was mapped in coding region (LOC_Os04g56170) of chromosome 4. RT-PCR revealed that the OsLG1 gene was not expressed liguleless mutants. Structure analysis of OsLG1 protein revealed that it predicted transcription factor with a highly conserved SBP domain consisting of 79 amino acids that overlapped a nuclear localization signal (NLS). RT-PCR revealed that OsLG1 is mainly expressed in vegetative organs.

• 한국균학회소식:학술대회논문집
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• 한국균학회 2018년도 춘계학술대회 및 임시총회
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• pp.37-37
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• 2018

Mating type of Lentinula edodes is determined by two unlinked genetic loci, A and B. To better understand mating behavior of L. edodes, we investigated variations in mating type genes in129 dikaryotic strains collected from East Asia. Through sequence analysis of A locus, we discovered that hypervariable region spanning N-term of HD2-intergenic region-N-term of HD1 could represent A mating type. Mating and hypervariable region analyses revealed 70 unique A mating types: 27 from 98 cultivated strains, 53 from 31 wild strains, and 10 commonly found. It was also revealed that only a few A mating type alleles such as A1, A4, A5, and A7 were prevalent in cultivated strains. Contrarily, A mating type in wild strains was highly diverse: 23 unique A alleles were discovered in small mountainous area in Korean peninsula, suggesting rapid evolution of A mating type in nature. The B locus was assessed by allelic variations in pheromone (PHB) and pheromone receptor (RCB) pairs which constituted subloci Ba and Bb. Sequence analyses and mating assay revealed 5 alleles of RCB1 with 9 associated PHBs in Ba sublocus and 3 alleles of RCB2 with 5 associated PHBs in Bb sublocus. Each RCB was primarily associated with two PHBs. Each PHB-RCB pair was always discovered as a distinct unit. This allowed us to propose 15 B mating types via combinations of five Ba and three Bb subloci. Further investigation on 129 strains confirmed that the B locus, unlike the A locus, was indeed restricted to 15 mating types. Thus, the total number of mating types became 1,050 in L. edodes through a combination of 70 A and 15 B. This number will further increase because of rapid diversification of A mating type. Our findings provide a comprehensive and practical knowledge on mating behaviors of L. edodes.