• Journal of Microbiology and Biotechnology
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• 제15권2호
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• pp.388-394
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• 2005

In order to investigate the distribution of dominant Bifidobacterium species in intestinal microflora of Korean adults and seniors, SDS-PAGE profiles of whole cell proteins were used for the identification of bifidobacteria. To confirm the reliability of SDS-PAGE, the Bifidobacterium species identified by SDS-PAGE of whole cell proteins were validated by using 16S rDNA sequencing analysis. The results of SDS­PAGE corresponded well with those determined by the analysis of 16S rDNA sequencing. Based on the analysis of SDS-PAGE patterns on unidentified fecal strains which showed positive in fructose-6-phosphate phosphoketolase activity, B. adolescentis, B. longum, and B. bifidum were identified in the feces of adults, and B. adolescentis, B. longum, B. bifidum, B. breve, and B. dentium were identified in those of seniors. In most of the fecal samples tested, the predominant Bifidobacterium species consisted of only a few species, and differences in the distribution and numbers of Bifidobacterium species were observed between adults and seniors. B. adolescentis and B. longum were found to be the most common species in feces of adults, but not in seniors. Accordingly, the distribution and abundance of bifidobacteria in the human intestinal microflora varied depending on the age of hosts.

• 대한한의학회지
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• 제32권5호
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• pp.41-49
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• 2011

Objectives: To optimize the preservation method of herbal decoction, we investigated the content of principle components of Pyungwi-san, liquiritin, glycyrrhizin, and hesperidin according to preservation temperature and period. We also investigated the changing patterns of pH and microbial population in Pyungwi-san decoction as a model case. Methods: With samples preserved at different temperatures, the content of liquiritin, glycyrrhizin, and hesperidin was determined using HPLC and microbial population was determined as viable counting method up to 8 times every month. Identification of isolated bacteria was performed by 16S rDNA analysis. Results: The content of liquiritin and glycyrrhizin did not change according to the preservation temperature and period, but that of hesperidin was severely decreased at room temperature. The isolate from the decoction was identified as Bacillus licheniformis by 16S rDNA sequence analysis. Microbial population appeared after 3 months' preservation and reached maximum value at 4 months; at all tested temperatures, the pH showed the lowest value (4.4-4.5) simultaneously. Conclusion: From the results, it seems to be that the microbial growth affects the pH of preserved decoction but not the change of liquiritin and glycyrrhizin content.

• KSBB Journal
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• 제26권5호
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• pp.407-411
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• 2011

Microbial fuel cell (MFC) wes enriched using sludge in wastewater treatment. The microbial community of activated sludge and enriched MFC were analyzed by FISH (fluorescent in situ hybridization) and 16S rDNA sequencing. Bacteroidetes group were pre-dominant in activated sludge by FISH. ${\alpha}$ group, ${\gamma}$ group and Acintobacter group were dominant and they were similar to distribution. The average value of 10 peak of MFC is 0.44C. When MFC wase enriched by sludge, ${\gamma}$-Proteobacteria, Plantomycetes group increased 70% and 60%, respectively. In results of 16S rDNA sequencing, Sphiringomonas sp. was comprised in ${\alpha}$ proteobacteria and Enterobacter sp., Klebsiella sp., Acinetobacter sp., Bacillus sp. were comprised in ${\gamma}$ proteobacteria and Chryseobacterium sp. was comprised in Flavobacteria were isolated from sludge.

• Journal of Animal Science and Technology
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• 제47권3호
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• pp.465-474
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• 2005

A two-step broad-range PCR method detecting gram-negative bacteria at the level as low as 2 CFU was developed by using primers of GNFI and GNRI and then semi-nested primer of GNF2 and GNRI. The nucleotide sequences of the primers were determined based on l6S rRNA gene. The DNA fragments of 1173 bp and 169 bp were amplified in one-step PCRs with primer sets of GNFI-GNRI and GNF2-GNRl, respectively, using template DNA from seven strains of gram-negative bacteria including Escherichia coli, Enterobacter aerogenes, Klebsiella pneumoniae, Pseudomonas spp., and Acinetobacter baumaii but not from Achromobacter lyticus, Alca/igens faecalis, and five strains of gram-positive bacteria. DNA fragments of 180 bp were amplified from LTLT-pasteurized milk and UHf-pasteurized milk in the two-step PCR. The DNA fragments were amplified from LTLT-pasteurized milk which was added with Pseudomonas j/uorescens and subsequently heated at 65 $^{\circ}C$, 80 $^{\circ}C$, and 100 $^{\circ}C$ for 30 min but they were not amplified from the milk autoclaved at 121$^{\circ}C$ for 15 min. It was suggested in PCR that Pseudomonas fluorescens heated at 65 $^{\circ}C$ for 30 min in milk was more sensitive to DNase treatment than viable bacteria.

• 한국식품저장유통학회지
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• 제17권3호
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• pp.410-417
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• 2010

우리나라 전통발효식품의 하나인 청국장으로부터 고 비활성 세포외 protease 생산능이 우수한 균주를 분리하고 그 특성을 조사하였다. 분리된 균주 중 D14 균주 배양 상징액의 protease 활성이 15.2 U/mL로서 가장 강하였으며 비활성 또한 40.0 U/mg protein으로서 가장 강하게 나타났다. 이 균주의 특성을 조사한 후 Berge's Manual of Systematic Bacteriology에 준하여 Bacillus subtilis로 동정하였다. 균주 D14의 16S rDNA 염기서열을 분석하여 B. subtilis subsp. subtilis NCIB 3610 균주와 99.9%의 상동성을 가짐을 확인 하였다. 또한 16S rDNA의 염기서열을 토대로 계통분석을 통하여 다른 Bacillus sp.의 균주들보다 NCIB 3610 균주와 유전적으로 매우 가까운 관계에 있음을 확인하였다. Soluble starch는 사용한 탄소원 중 가장 높은 수준의 protease 생산능을 보였으나 단당류, 이당류 등은 모두 효소 생산능에 대하여 저해효과를 나타내었다. 특히 fructose를 첨가한 경우에는 12.7%, glucose를 첨가한 경우에는 35.9% 수준의 효소 생산능을 나타내었다. 질소원의 경우는 soytone의 첨가구에서 대조구에 비하여 약 61.4% 수준의 효소활성을 나타내어 저해효과가 가장 높았으며 다른 질소원은 효소 생산에 크게 영향을 미치지 않았다.

• 생태와환경
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• 제47권4호
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• pp.342-349
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• 2014

본 연구는 최근 북한강 수계에서 번성하고 있는 Anabaena strain의 16S rDNA 염기서열을 이용한 종 수준의 동정과 geosmin 합성 유전자의 탐침을 통해 이취미 물질의 잠재 생산능력을 분석하였다. 현장(경기도 양평군 조암면 삼봉리 수역)에서 분리 배양한 Anabaena는 직선형과 나선형 두 가지의 형태적 변이를 보였다. 이들은 세포의 크기와 사상체에서 형태적 차이를 나타냈으며, A. circinalis 및 A. crassa와 유사한 형태적 특징들을 보여주었다. 그러나 16S rDNA 계통수 및 유연관계를 분석한 결과, 직선형과 나선형 모두 동일한 A. circinalis 종으로 확인되었다(98%~100%의 유전적 유사도). 또한 직선형과 나선형 strain 모두에서 geosmin을 합성하는 유전자 구간이 발견되어, 북한강 수계에 존재하는 Anabaena circinalis는 종의 형태적 변이에 관계없이 geosmin을 생산할 수 있음을 보여주었다. 본 연구의 결과는 유전자 수준에서 A. circinalis의 geosmin 생산에 대한 직접적인 증거를 제공하는 국내 최초의 보고로서 북한강 수계에서 geosmin 증가의 원인종 확인 및 관리에 중요한 자료를 제공한다.

• Molecules and Cells
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• 제25권4호
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• pp.510-522
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• 2008

To construct the molecular systematics of the genus Megoura (Hemiptera: Aphididae), DNA based-identification was performed using four mitochondrial and three nuclear DNA regions: partial cytochrome c oxidase I (COI), partial tRNA-leucine + cytochrome c oxidase II (tRNA/COII), cytochrome b (CytB), partial 12S rRNA + tRNA-valine + 16S rRNA (12S/16S), elongation factor-1 alpha ($EF1{\alpha}$), and the internal transcribed spacers 1 and 2 (ITS1, ITS2). Pairwise sequence divergences between taxa were compared, and phylogenetic analyses were performed based on each DNA region separately, and the combined datasets. COI, CytB, $EF1{\alpha}$, ITS1, and ITS2 were relatively effective in determining species and resolving their relationships. By contrast, the sequences of tRNA/COII and 12S/16S were not able to separate the closely related species. CytB and $EF1{\alpha}$ gave better resolution with higher average sequence divergences (4.7% for CytB, 5.2% for $EF1{\alpha}$). The sequence divergence of COI (3.0%) was moderate, and those of the two ITS regions (1.8% for ITS1, 2.0% for ITS2) were very low. Phylogenetic trees were constructed by minimum evolution, maximum parsimony, maximum likelihood, and Bayesian phylogenetic analyses. The results indicated that the phylogenetic relationships between Megoura species were associated with their host preferences. Megoura brevipilosa and M. lespedezae living on Lespedeza were closely related, and M. nigra, monophagous on Vicia venosa, was rather different from M. crassicauda, M. litoralis, and M. viciae, which are oligophagous on Lathyrus and Vicia. The three populations of M. crassicauda formed a clade separated from M. litoralis and M. viciae. Nevertheless M. litoralis and M. viciae, which are morphologically similar, were not separated due to negligible sequence divergence. We discuss the phylogenetic relationships of the Megoura, and the usefulness of the seven DNA regions for determining the species level phylogeny of aphids.

• Animal cells and systems
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• 제16권5호
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• pp.415-424
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• 2012

Four species of ophichthid leptocephali were identified using 12S rDNA sequences, and their morphological descriptions were first provided based on six individuals (S1-S3,M1, and E1-E2) collected from the East Sea and the Korea Strait between September 2008 and October 2010. Mitochondrial 12S rDNA 859-861 base pairs of ophichthid leptocephali were compared with those of 16 ophichthids adult and 2 outgroups (Anguilla japonica and Conger myriaster). Leptocephali of S1 and E1 were very closely clustered with adult of Scolecenchelys borealis (D=0.002) and Echelus uropterus (D=0.000), respectively. However, leptocephali of S2-S3 andM1 were slightly far clustered with leptocephalus of S1 (D=0.006) and adult of Muraenichthys gymnopterus (0.034), respectively. We believe that S1 and E1 are S. borealis and E. uropterus, respectively, in which the former is unrecorded species in Korea. However, S2-S3 and M1 may be undescribed species belonging to genus Scolecenchelys and Muraenichthys, respectively, because total numbers of myomeres for S2-S3 (148-158) and M1 (151) were not consistent with total numbers of vertebrae or distribution for any adult of Scolecenchelys spp. and Muraenichthys spp. in the world. We propose the new Korean name 'Dong-hae-mul-baem' for S. borealis.

• Parasites, Hosts and Diseases
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• 제43권1호
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• pp.7-13
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• 2005

The taxonomy of Acanthamoeba spp., an amphizoic amoeba which causes granulomatous amoebic encephalitis and chronic amoebic keratitis, has been revised many times. The taxonomic validity of some species has yet to be assessed. In this paper, we analyzed the morphological characteristics, nuclear 18s rDNA and mitochondrial 16s rDNA sequences and the Mt DNA RFLP of the type strains of four Acanthamoeba species, which had been previously designated as A. divionensis, A. parasidionensis, A. mauritaniensis, and A. rhysodes. The four isolates revealed characteristic group II morphology. They exhibited 18S rDNA sequence differences of $0.2-1.1\%$ with each other, but more than $2\%$ difference from the other compared reference strains. Four isolates formed a different clade from that of A. castellanii Castellani and the other strains in morphological group lion the phylogenetic tree. In light of these results, A. paradivionensis, A. divionensis, and A. mauritaniensis should be regarded as synonyms for A. rhysodes.

• 한국미생물·생명공학회지
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• 제35권4호
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• pp.347-351
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• 2007

감자 더뎅이병은 제주도 전 지역에서 발생하는 병해로서 더뎅이병 발생 시 경제적인 손실이 막대한 것으로 알려져 있다. 본 연구에서는 제주도 감자 더뎅이병징이 있는 부위에서 Streptomyces spp.를 분리, 배양한 후, 16S rRNA 유전자를 이용하여 계통분석을 실시하였다. 계통분석 결과 제주도 감자 더뎅이병징이 있는 부위에서 분리된 균은 모두 Streptomyces spp.에 속하였으며, 대부분이 기존에 더뎅이병을 일으키는 Streptomyces spp.로 확인되었다. 그러나 일부의 분리균은 기존에 알려진 감자 더뎅이병원균과는 다르다. 따라서 이들 병원균에 대해서는 지방산과 단백질 분석, 그리고 DNA-DNA 혼성화 등과 같은 보다 많은 연구의 수행을 통하여 새로운 더뎅이병을 일으키는 Streptomyces spp.인지, 또는 아직까지 명명되지 않은 Streptomyces spp.인지를 확인해야 할 것으로 사료된다.