• 식물병연구
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• 제11권1호
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• pp.56-65
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• 2005

이 연구는 Alternaria속 균류 중에서 분생포자의 형태가 유사하여 분류, 동정이 매우 어려운 소형의 포자를 형성하는 11종의 Alternaria의 종간 유연관계와 분류체계를 확립하기 위하여 공시균들의 ribosomal DNA의 ITS 및 미토콘드리아 small submit 영역의 염기서열 분석, 그리고 URP primer에 의한 핵산지문분석을 실시하였다. 소형의 포자를 형성하는 11종 Alternaria속의 rDNA internal transcribed spacer(ITS) 영역과 미토콘드리아 small submit rDNA 의 염기서열을 분석하였던 바 A. infectoria를 제외한 10종의 Alternaria에서 100%의 상동성을 나타내었다. 이는 공시한 10종의 Alternaria가 유전적으로 매우 근연의 관계에 있음을 나타내며, 이 marker는 공시균들의 종구분에 이용할 수 없음을 나타내는 것이다. URP primer 10종을 공시하여 소형의 포자를 형성하는 Alternaria 균류의 핵산지문분석을 실시한 결과, 개개의 primer 로는 종간의 구분이 불가능하였으나 여러개의 primer를 종합하여 UPGMA분석을 실시할 경우 비록 종간 유사도는 높았디만 각각의 종은 독립된 cluster를 형성하여 종간 구별이 가능하였다. 자리공에서 분리한 Alternaria sp.는 URP-PCR 핵산지문 분석 결과 다른 Alternaria 종들과 차이가 인정됨으로 이 균은 미기록인 신종의 Alternaria로 사료되었다. A.infectoria는 다른 Alternaria 종들과 ITS 분석 및 URP-PCR 핵산지문분석에서 큰 차이를 보임으로서 뚜렷이 구별되는 종으로 판단되었다.

• 미생물학회지
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• 제35권3호
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• pp.180-184
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• 1999

Agaricus 속의 계통분류학적 유연관계를 알아보기 위하여 Internal transcribed spacers 1,2, 와 5.8S rDNA 에 해당하는 부위를 PCR 기법으로 증폭하여 염기서열을 비교 조사하였다. 본 실험에 사용되어진 aGARICUS 속 5균주에 대한 염기서열을 분석한 결과, 이들 속은 크게 두 개의 군으로 분류되었다. 첫 번째 분류군은 aGARICUS BLAZEI atcc 76739, 현재 우리나라에서 배양중인 Agaricus blazei, agaricus arvensis IMSNU 32049, 그리고 Agaricus campestris VPI-OKM 25665로 이루어 졌다. 특히, Agaricus blazei ATCC 76739와 현재 우리나라 농가에서 배양중인 Agaricus blazeisms ITS 부위에서 5개의 염기서열에서 변이가 발견되었다. 이러한 변이는 지리적 또는 배양상의 변이로 인하여 특정염기서열에서 변이가 발생한 것으로 간주되므로 이들은 상호 동일종일 것으로 추정된다. 또한, Agaricus arvensis IMSNU 32049 와 Agaricus ampestris VPI-LKM 25665는 Agaricus blazei 하위 분류군을 형성하였다. 두 번째 분류군은 Agaricus bisporus CH 3004와 Agaricus pocillator DUKE-J 173 으로 이루어졌다.

• Journal of Microbiology and Biotechnology
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• 제5권4호
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• pp.194-199
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• 1995

A portion (7.4 kb) of ribosomal DNA tandem repeat unit from a genome of the mushroom T. matsutake has been cloned. A 1.75 kb EcoRI fragment was cloned first using S. cerevisiae 255 rRNA gene as a probe, and this was then used for further cloning. A chromosomal walking experiment was carried out and the upstream region of the 1.75 kb fragment was cloned using SmaI/BamHI enzyme, the size was estimated to be 5.2 kb in length. Part of the downstream region of the 1.75 kb fragment was also cloned using XbaI/BamHI enzymes. Restriction enzyme maps of three cloned DNA fragments were constructed. Northern hybridization, using total RNA of T. matsutake, and the restriction fragments of three cloned DNAs as probes, revealed that all four ribosomal RNA genes (large subunit[LSU], small subunit [SSU], 5.85 and 5S rRNA genes) are present in the cloned region. The gene organization of the rDNA are regarded as an intergenic spacer [IGS]2 (partial) - SSU rRNA - internal transcribed spacer [ITS]1 - 5.8S rRNA - ITS2 - LSU rRNA - IGS1 -5S rRNA - IG52 (partial).

• Bulletin of the Korean Chemical Society
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• 제35권10호
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• pp.3005-3008
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• 2014

Human ChlR1 protein (hChlR1), a member of the cohesion establishment factor family, plays an important role in the segregation of sister chromatids for maintenance of genome integrity. We previously reported that hChlR1 interacts with hFen1 and stimulates its nuclease activity on the flap-structured DNA substrate covered with RPA. To elucidate the relationship between hChlR1 and Okazaki fragment processing, the effect of hChlR1 on in vitro nuclease activities of hFen1 and hDna2 was examined. Independent of ATPase activity, hChlR1 stimulated endonuclease activity of hFen1 but not that of hDna2. Our findings suggest that the acceleration of Okazaki fragment processing near cohesions may aid in reducing the size of the replication machinery, thereby facilitating its entry through the cohesin ring.

• Mycobiology
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• 제38권3호
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• pp.166-170
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• 2010

Gummy stem blight is a major foliar disease of muskmelon (Cucumis melo L.). In this study, morphological characteristics and rDNA internal transcribed spacer (ITS) sequences were analyzed to identify the causal organism of this disease. Morphological examination of the Jeonbuk isolate revealed that the percentage of monoseptal conidia ranged from 0% to 10%, and the average length $\times$ width of the conidia was 70 ($\pm$ 0.96) $\times$ 32.0 ($\pm$ 0.15) ${\mu}m$ on potato dextrose agar. The BLAST analysis showed nucleotide gaps of 1/494, 2/492, and 1/478 with identities of 485/492 (98%), 492/494 (99%), 491/494 (99%), and 476/478 (99%). The similarity in sequence identity between the rDNA ITS region of the Jeonbuk isolate and other Didymella bryoniae from BLAST searches of GenBank was 100% and was 95.0% within the group. Nucleotide sequences of the rDNA ITS region from pure culture ranged from 98.2% to 99.8%. Phylogenetic analysis with related species of D. bryoniae revealed that D. bryoniae is a monophyletic group distinguishable from other Didymella spp., including Ascochyta pinodes, Mycosphaerella pinodes, M. zeae-maydis, D. pinodes, D. applanata, D. exigua, D. rabiei, D. lentis, D. fabae, and D. vitalbina. Phylogenetic analysis, based on rDNA ITS sequence, clearly distinguished D. bryoniae and Didymella spp. from the 10 other species studied. This study identified the Jeonbuk isolate to be D. bryoniae.

• ALGAE
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• 제38권4호
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• pp.241-252
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• 2023

The genera Carolibrandtia and Chlorella have been described as small green algae with spherical cell shapes that inhabit various environments. Species of these genera are often difficult to identify because of their simple morphology and high phenotypic plasticity. We investigated two small coccoid strains from Antarctica based on morphology, molecular phylogeny by two alignment methods which have been applied to previous phylogenetic studies of the genus Chlorella, and comparison of the secondary structures of nuclear small subunit (SSU) and internal transcribed spacer (ITS) rDNA sequences. Light microscopy of two strains revealed spherical cells containing chloroplasts with pyrenoids, and the morphological characteristics of the strains were nearly identical to those of other Chlorella species. However, based on the phylogenetic analyses of nuclear SSU and ITS rDNA sequences, it was determined that the Antarctic microalgal strains belonged to two genera, as the Chlorella and Carolibrandtia. In addition, the secondary structures of the SSU and ITS2 sequences were analyzed to detect compensatory base changes (CBCs) that were used to identify and describe the two strains. A unique CBC in the SSU rDNA gene was decisive for distinguishing strain CCAP 211/45. The ITS2 rDNA sequences for each strain were compared to those obtained previously from other closely related species. Following the comparison of morphological and molecular characteristics, we propose KSF0092 as a new species, Chlorella terrestris sp. nov., and the reassignment of the strain Chlorella antarctica CCAP 211/45 into Carolibrandtia antarctica comb. nov.

• 한국한의학연구원논문집
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• 제14권1호
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• pp.129-135
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• 2008

Phylogenetic relationship and DNA polymorphism among local populations of the Asparagus cochinchinensis have been investigated based on nuclear ribosomal DNA ITS sequences and RAPD analysis in Korea. In result, two genetically distinct groups of local populations except Geoje were recognized by the phylogenetic tree both in rDNA-ITS and RAPD. One was called 'western coast group' that includes the Buan 1, 2 and Taean and the other was 'southern coast group' that includes Haenam, Yeosu and Namhae. Thus, the geographical relationship of Asparagus cochinchinensis was two well-typified clades. These results suggest that the geographical genetic variation of Asparagus cochinchinensis is closely connected with the slow and long period of propagation via the coast in Korean Peninsula.

• 생명과학회지
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• 제13권1호
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• pp.99-104
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• 2003

Aromatic compounds are of major concern among environmental pollutants due to their toxicity and persistence. To monitor aromatic compounds in a real time with a better sensitivity, a new method of SPR (surface plasmon resonance) based on DNA chip (Biacore 3000) was developed here. It is thought that CapR regulatory protein as a complex with phenol, could bind to their corresponding promoter, Po. Biotinylated DNA oligomers for the promoter was synthesized by PCR and coupled onto streptoavidin-linked CM5-chip. CapR regulatory proteins were purified after cloning their genes in pET21a (+) vector and expressing proteins. The interaction was assessed by the system where the regulatory proteins flowed with or without phenol through the cells of DNA chip. CapR regulatory protein even in the presence of phenol had no response to its promoter, Po, suggesting that other factor(s) might be required for the activation of Po promoter. The present work reveals a promising possibility of the SPR-based DNA chip in monitoring specific environmental pollutants in a real time.

• The Plant Pathology Journal
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• 제26권1호
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• pp.83-89
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• 2010

Taxonomic position of 46 Korean isolates of Rhizoctonia solani which were classified into nine intraspecific groups by anastomosis and cultural characteristics was analyzed by randomly amplified polymorphic DNA (RAPD) and sequence analyses of the internal transcribed spacer (ITS) regions of ribosomal DNA. All the isolates within each group showed highly similar band patterns in RAPD. The ITS regions of the isolates within the same groups showed a high level of sequence similarity above 96.0% whereas similarities among different groups were below 94.4%. When compared with several reference strains of R. solani from foreign countries, all the Korean isolates were clustered with the foreign isolates belonging to the same groups in the phylogenetic tree. All six Korean strains of AG-4 were identified as HG-1 out of 3 subgroup of AG-4. We discussed taxonomic position of Korean isolates of R. solani and showed that sequence analysis with ITS regions could be a rapid and useful method for identification of intraspecific group of R. solani.

• 생명과학회지
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• 제13권4호
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• pp.400-411
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• 2003

5.8S rDNA를 포함한 ITS부위에 대한 염기서열 분석결과, 종에 따라 다양한 염기서열을 가지고 있어 분류에 이용될 수 있었으며, 특히 ITS2부위보다 ITS1부위에서 종에 대한 변이율이 높은 것으로 나타났다. 아울러 균종에 따라 정도의 차이는 있으나 사용된 모든 종들이 서로 계통분류학적 거리가 멀어서 종간의 구분이 명확하게 나타났다. P. tenuipes, I. japonica, P. japonicus는 multiple alignment분석에서 매우 유사한 염기서열을 가지고 있어, 이들 세종은 같은 종이지만 다른 이름으로 불리고 있는 것으로 나타났으며, 아울러 Paecilomyces sp. KACC 40220과 KACC 40656도 동일한 염기서열을 가지고 있어 p. tenuipes로 판단된다. 국내에서 유통되는 동충하초제품 35건과 중국산 1건에 대해 실험한 결과 23건은 P. tenuipes / japonica로, 11건은 C. militaris로, 1건은 B. bassiana로 분류되었으며, 중국산 제품 1건은 C. multiaxialis로 분류되었다.