• International Journal of Oral Biology
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• 제36권1호
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• pp.1-6
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• 2011

The purpose of this study was to develop species-specific real-time quantitative PCR (RT-qPCR) primers for use in the detection of Aggregatibacter actinomycetemcomitans. These primers were designed based on the nucleotide sequences of the RNA polymerase ${\beta}$-subunit gene (rpoB). We assessed the specificity of the primers against nine strains of A. actinomycetemcomitans, eight strains (three species) of the Haemophilus genus, and 40 strains of 40 other oral bacterial species. Primer sensitivity was determined by testing serial dilutions of the purified genomic DNAs of A. actinomycetemcomitans ATCC $33384^T$. Our data reveal that we had obtained species-specific amplicons for all of the tested A. actinomycetemcomitans strains, and that none of these amplicons occurred in any of the other species. Our PCR protocol proved able to detect as little as 2 fg of A. actinomycetemcomitans chromosomal DNA. Our findings suggest that these qRT-PCR primers are suitable for application in epidemiological studies.

• 한국축산식품학회지
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• 제35권1호
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• pp.1-9
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• 2015

In recent years, the knowledge about bifidobacteria has considerably evolved thanks to recent progress in molecular biology. The analysis of the whole genome sequences of 48 taxa of bifidobacteria offers new perspectives for their classification, especially to set up limit between two species. Indeed, several species are presenting a high homology and should be reclassified. On the other hand, some subspecies are presenting a low homology and should therefore be reclassified into different species. In addition, a better knowledge of the genome of bifidobacteria allows a better understanding of the mechanisms involved in complex carbohydrate metabolism. The genome of some species of bifidobacteria from human but also from animal origin demonstrates high presence in genes involved in the metabolism of complex oligosaccharides. Those species should be further tested to confirm their potential to metabolize complex oligosaccharides in vitro and in vivo.

• 한국미생물학회:학술대회논문집
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• 한국미생물학회 2008년도 International Meeting of the Microbiological Society of Korea
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• pp.106-106
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• 2008

• 한국균학회지
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• 제30권2호
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• pp.78-85
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• 2002

저장배에서 분리한 15그룹 3종을 KCTC, KCCM, 충남대에서 분양받은 16균주를 대상으로 상호 유연관계를 분석하고자 배양적 특성을 관찰하고 RAPD 검정에 의한 진단 가능성을 검토하였다. 병원균의 배양특성을 조사한 결과 배지상 생육형태, 색, 속도 등에 있어서 균주간 차이를 보였으며 배양적 특성과 RAPD에 의한 유연관계 분석 결과를 비교 분석하기 위하여 배에서 분리한 15개 균주와 16개 표준 균주의 종간 다형성을 관찰하였다. RAPD을 이용하여 유전적 다양성을 분석하기 위하여 5개의 URP primer를 이용하여 genomic DNA를 증폭시킨 결과 $0.1{\sim}2.0kb$ 크기의 총 744개 band들이 증폭되었다. cluster 분석결과 전체 유사도는 47.7%로 31 균주들은 4개의 genomic DNA RAPD 집단으로 분류되었다. 배에서 분리한 P. expansum 균주간에는 87.4% 이상의 상동성을 보였고, P. solitum 균주간에는 97.5%, P. crustosum 균주 간에는 95.1% 이상의 상동성을 보였다. 배에서 분리된 동일한 종내에서는 배양적 특성에 의한 동정결과와 RAPD에 의한 분석결과와 일치하였다.

• 한국수산과학회지
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• 제54권4호
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• pp.489-497
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• 2021

Climate change is affecting the evolutionary trajectories of individual species and ecological communities, partly through the creation of new species groups. As population shift geographically and temporally as a result of climate change, reproductive interactions between previously isolated species are inevitable and it could potentially lead to invasion, speciation, or even extinction. Four species of abalone, genus Haliotis are present along the Korean coastline and these species are important for commercial and fisheries resources management. In this study, genetic markers for fisheries resources management were discovered based on genomic information, as part of the management of endemic species in response to climate change. Two thousand one hundred and sixty one single nucleotide polymorphisms (SNPs) were discovered using genotyping-by-sequencing (GBS) method. Forty-one SNPs were selected based on their features for species classification. Machine learning analysis using these SNPs makes it possible to differentiate four Haliotis species and hybrids. In conclusion, the proposed machine learning method has potentials for species classification of the genus Haliotis. Our results will provide valuable data for biodiversity conservation and management of abalone population in Korea.

• 환경생물
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• 제36권3호
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• pp.352-358
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• 2018

본 연구는 PCR 기반 RFLP (Restriction Fragment Length Polymorphism; 제한절편 길이 다형성) 분자기법을 활용하여 난 및 치어 대상 납자루아과 어류 3종의 동정을 좀 더 빠르고 정확하게 파악하고 납자루아과 어류의 종별 산란양상 및 번식생태 이해에 대한 기여가 목적이다. 본 연구를 위해 기존 선행된 문헌자료를 확인하고 납자루아과 어류가 2종 이상 동서하고 있는 지역을 확인하여 현지조사를 수행하였다. 현지조사 결과 확인된 납자루아과 어류는 묵납자루(Acheilognathus signifer), 줄납자루(A. yamatsutae) 및 각시붕어(Rhodeus uyekii)로 총 3종이 확인되었으며, 확인된 납자루아과 어류와 동서하고 있는 숙주조개(작은말조개; Unio douglasiae sinuolatus)를 채집하여 숙주조개 속 납자루아과 어류의 난 및 치어를 확보하였다. 현지조사 결과 확인된 납자루아과 어류 3종을 대상으로 미토콘드리아 DNA COI과 cyt b 유전자 염기서열을 비교하여 각각 종별로 특이성을 지닌 부위(단일염기변이; Single Nucleotide Variation: SNV)에 맞는 제한효소를 선정하였고, 숙주조개 속 난 및 치어를 대상으로 genomic DNA를 추출하여 PCR-RFLP 실험을 수행한 결과 현지조사 시 확인된 납자루아과 어류 3종의 독특한 제한절편 길이 양상을 전기영동을 통하여 확인하였다. 본 연구를 통해 묵납자루, 줄납자루 및 각시붕어의 종을 판별할 수 있는 RFLP 마커를 개발하였으며, 숙주조개 난 및 치어를 대상으로 정확한 종의 동정을 보다 빠르고 효과적으로 수행하여 각각 납자루아과 종별 산란양상을 보다 정확히 규명하고 향후 이들 자연개체군의 효과적인 유지, 관리 및 보전 방법 개발에 유용하게 활용될 수 있을 것으로 판단된다.

• 한국수산과학회지
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• 제44권2호
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• pp.126-140
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• 2011

Genetic determinants of two metallothionein isoforms (MT-A and MT-B) were isolated and characterized from the perciform species, rockbream (Oplegnathus fasciatus). Rockbream MT-A and MT-B shared a high degree of homology at amino acid levels with representative orthologs from other perciform species, especially with respect to the conserved cysteine residues. At the genomic level, both MT-A and MT-B genes represent a tripartite structure typical of vertebrate MT genes. However, rockbream MT-B showed unusually large introns (1.2 kb and 0.8 kb for intron I and II, respectively), a phenomenon that has rarely been seen in other vertebrate MT genes. MT-A and MT-B transcripts were ubiquitously detected in a wide array of tissues, wherein brain and eye showed the highest basal expression levels, and the fin exhibited the lowest expression of both isoforms. The basal expression of MT-A in most tissues was significantly higher (ranging from 4- to 10-fold) than that of MT-B. Upon heavy metal exposures to Cd, Cu or Zn at 25 ppb for 48 h, MT-A and MT-B transcripts in the liver were significantly activated by Cd and moderately by Zn. On the other hand, exposure to Cu did not result in alterations of MT-A, nor in the significant suppression of MT-B. Following bacterial challenges with Escherichia coli, Edwardsiella tarda or Streptococcus iniae, MT isoforms in the liver, kidney and spleen were highly modulated and exhibited a pattern that was dependent on the bacterial species, tissues and isoforms. These results suggest that the two MT isoforms could be taken into account as potential indicators of metal toxicity and immune perturbations of this aquaculture-relevant species.

• Journal of Microbiology and Biotechnology
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• 제27권8호
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• pp.1419-1427
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• 2017

As probiotics play an important role in maintaining a healthy gut flora environment through antitoxin activity and inhibition of pathogen colonization, they have been of interest to the medical research community for quite some time now. Probiotic bacteria such as Lactobacillus plantarum, which can be found in fermented food, are of particular interest given their easy accessibility. We performed whole-genome sequencing and genomic analysis on a GB-LP1 strain of L. plantarum isolated from Korean traditional fermented food; this strain is well known for its functions in immune response, suppression of pathogen growth, and antitoxin effects. The complete genome sequence of GB-LP1 is a single chromosome of 3,040,388 bp with 2,899 predicted open reading frames. Genomic analysis of GB-LP1 revealed two CRISPR regions and genes showing accelerated evolution, which may have antibiotic and antitoxin functions. The aim of the present study was to predict strain specific-genomic characteristics and assess the potential of this new strain as lactic acid bacteria at the genomic level using in silico analysis. These results provide insight into the L. plantarum species as well as confirm the possibility of its utility as a candidate probiotic.

• 대한수의학회지
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• 제38권4호
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• pp.771-776
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• 1998

Synthetic oligonucleotide primers of 20 and 21 bases, respectively, were used in the polymerase chain reaction (PCR) to amplify a sequence of the mrp gene, which encodes the muramidase released protein of Streptococcus suis. Amplification was not recorded when 5 other streptococcal species were tested or when 9 different nonstreptococcal species were tested. A DNA fragment of 517bp was amplified from the genomic DNA of S suis. The lower detection limit was 100pg of the genomic DNA. The primers recognized 34 serotypes of S suis reference strains and 9 isolates from pneumonic lung, brain, nasal discharge, tonsil. This results suggest that the amplification of the mrp gene by PCR method is potential for the identification of S suis isolates.

• Animal cells and systems
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• 제13권3호
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• pp.351-356
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• 2009

Ciliates are undoubtedly one of the most diverse protozoans that play a significant role in ecology. However, molecular examination, based on comparing the DNA sequences, has been done on a limited number of the species. Because most ciliates are uncultivable and their population sizes are often too small, it is usually difficult to obtain sufficient genomic DNA required for PCR based experiments. In the present study, we evaluated the effectiveness of four commercial DNA extraction procedures that extract high quality genomic DNA from a single ciliate cell. It was discovered that RED Extract-N-$Amp^{TM}$ PCR kit is the best method for removing PCR-inhibiting substances and minimizing DNA loss during purification. This method can also amplify more than 25 reactions of PCR. In addition, this technique was applied to single cells of 19 species belonged to 7 orders under 5 classes that isolated from mixed natural populations. Their small subunit ribosomal DNA (SSU rDNA) was successfully amplified. In summary, we developed a simple technique for the high-yield extraction of purified DNA from a single ciliate cell that may be more useful for rare ciliates, such as tiny and uncultivable marine microbes.