• 한국균학회지
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• 제26권2호통권85호
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• pp.135-143
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• 1998

CHEF (Contour-Clamped homogeneous electric field) gel electrophoresis를 이용하여 Fusarium section Sporotrichiella, Liseola, Gibbosum, Discolor와 Martiella에 속하는 10종의 electrophoretic karyotype을 비교하였다. Intact chromosomal DNA는 균류의 원형질체로부터 추출하였으며, 크기에 따라 다양한 조건을 주어 DNA 분자를 분리시켰다. Fusarium속에 속하는 종의 염색체는 0.78Mb에서 7.20Mb의 크기를 가진 염색체가 종에 따라 $5{\sim}13$개였다. 각 종의 total genome 크기는 18.32Mb에서 48.20Mb였다. Electrophoretic karyotype을 비교한 후 F. oxysporum formae speciales lilii로부터 무작위로 선택하여 만든 genomic DNA를 probe로 하여 Southern hybridization 분석을 수행하였다.

• KSBB Journal
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• 제25권2호
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• pp.199-204
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• 2010

Drag-tag으로 사용될 반복단위 단백질을 생물학적인 방법을 통해 생산함으로써 수용액 내에서 DNA 분리가 가능함을 확인하였다. 서로 다른 크기를 갖는 두 종류의 반복단위 단백질을 디자인하였고, 이를 발현시킨 뒤 정제하였다. 정제된 반복단위 단백질에 형광 dye를 포함하고 있는 100 base의 DNA를 연결하였고, 이 연결 물질을 모세관 내부가 수용액으로 충진된 microchip 상에서 전기영동 하였다. 그 결과 생물학적으로 생산된 반복단위 단백질이 SNP 분석과 같은 빠르고 효율적인 DNA 분석에 적합한 후보물질로 사용될 수 있음을 확인하였다.

• BioChip Journal
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• 제12권4호
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• pp.340-347
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• 2018

$F{\ddot{o}}rster$ resonance energy transfer (FRET) is extremely sensitive to the separation distance between the donor and the acceptor which is ideal for probing such biological phenomena. Also, FRET-based probes have been developing for detecting an unamplified, low-abundance of target DNA. Here we describe the development of FRET based DNA sensor based on an accumulated QD system for detecting KRAS G12D mutation which is the most common mutation in cancer. The accumulated QD system consists of the polystyrene beads which surface is modified with carboxyl modified QDs. The QDs are sandwich-hybridized with DNA of a capture probe, a reporter probe with Texas-red, and a target DNA by EDC-NHS coupling. Because the carboxyl modified QDs are located closely to each other in the accumulated QDs, these neighboring QDs are enough to transfer the energy to the acceptor dyes. Therefore the FRET factor in the bead system is enhancing by the additional increase of 29.2% as compared to that in a single QD system. These results suggest that the accumulated nanobead probe with conjugated QDs can be used as ultrasensitive DNA nanosensors detecting the mutation in the various cancers.

• 대한의생명과학회지
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• 제16권3호
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• pp.193-196
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• 2010

Multiplex PCR-based short tandem repeat (STR) analysis is considered as a good tool for monitoring bone marrow engraftment after sex-mismatched allogeneic transplantation and provides a sensitive and accurate assessment of the contribution of both donor and/or recipient cells in post-transplantation specimens. Forensic STR analysis and quantitative real time PCR are used to determine the proportion of donor versus recipient each contained within the total DNA. The STR markers were co-amplified in a single reaction by using commercial $PowerPlex^{(R)}$ 16 system and $AmpFISTR^{(R)}$ $Identifiler^{(R)}$ / $Yfiler^{(R)}$ PCR amplification kits. Separation of the PCR products and fluorescence detection were performed by ABI $PRIS^{(R)}$ 3100 Genetic Analyzer with capillary electrophoresis. The $GeneMapper^{TM}$ ID software were used for size calling and analysis of STR profiles. Extracted DNA was quantified by the $Quantifiler^{TM}$ Human DNA / Y Human Male DNA Quantification Kit The intent of this study was to analyze the ratio of donor versus recipient cells in the post-transplant peripheral blood, spleen, lung and kidney specimens. Specimens were taken from the traffic accident male victim who had been engrafted from bone marrow female donor. Blood and spleen specimens displayed female donor DNA profile. Kidney specimen showed male recipient DNA profile. Interestingly, lung tissue showed mixed profiles. The findings of this study indicate that the forensic STR analysis using fluorescence labeling PCR combined with capillary electrophoresis is quick and reliable enough to assess the ratio of donor versus recipient cells and to monitor the mixed chimeric patterns.

• The Plant Pathology Journal
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• 제15권1호
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• pp.57-61
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• 1999

Xanthomonas campestris pv. glycines causes bacterial pustule disease on susceptible soybean leaves and produces a bacteriocin, named glycinecinA, against most xanthomonads including Xanthomonas campestris pv. vesicatoria. One of the 5 isolated DNA regions responsible for bacteriocin production, a 1.7 kb DNA region for the glycinecinA gene, was used as a probe to detect the presence of the homolog DNA in other bacterial strains. Among 55 bacterial strains tested, only X. campestris pv. glycines showed the positive signal with glycinecinA DNA. Two oligomers, heu2 and heu4, derived from a glycinecinA DNA were used to carry out the polymerase chain reaction (PCR) analysis with chromosomal DNA from 55 different bacterial strains including 24 different strains of X. campestris pv. glycines, 9 different pathovars of xanthomonads, and other 22 bacterial strains of different genus and species. By separation of the PCR products on agarose gel, a 0.86 kb DNA fragment was specifically detected when X. campestris pv. glycines was present in the amplification assay. The 0.86 kb fragment was not amplified when DNA from other bacteria was used for the assay. Southern analysis with glycinecinA DNA showed that the PCR signal was obtained with X. campestris pv. glycines isolates from various geographic regions and soybean cultivars. Therefore, the 1.7 kb DNA region for the glycinecinA gene can be used for the pathovar-specific probe for the DNA hybridization and the primers heu2 and heu4 can be used for the pathovar-specific primers for the PCR analysis to detect X. campestris pv. glycines.

• 식물분류학회지
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• 제45권2호
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• pp.158-168
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• 2015

한국산 물통이(Pilea)속 식물의 분자계통학적 연구를 통해서 총 1속 5분류군으로 정리하였다. 물통이속은 모두 1년생 초본으로, 그늘지고 습기가 있는 지역에서 서식하며, 여름에 꽃이 피고, 가을에 열매를 맺는다. nrDNA의 ITS regions과 cpDNA의 psbA-trnH regions의 DNA 염기서열의 분석 결과, 산물통이, 물통이는 분계조를 각각 형성하였다. 하지만 제주 산방산의 제주큰물통이는 내륙지역의 지리산에서 자생하는 제주큰물통이와 같은 분계조를 형성하지 못하고 큰물통이, 모시물통이와 섞여 분계조를 형성하였다. ITS1, 4 regions에서만 DNA 염기서열이 분석된 지리산의 제주큰물통이 역시 완전히 다른 분계조를 형성하였다. 단순히 지리적인 차이로 인해 형성되었다고 보기에 무리가 있을 것으로 생각되어지며, 후에 좀 더 많은 연구가 이루어져야 할 것으로 생각되어진다.

• Journal of Microbiology
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• 제41권2호
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• pp.157-160
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• 2003

To estimate genetic relationships within the genus Rhizopus, genetic variations in 20 strains were investigated by DGGE and PCR-RFLP of rDNA ITS region (ITSI, ITS2,5.8S). The size of the amplified products showed the interspecific polymorphisms, 650 bp,700 bp, and 900 bp. The DGGE approach allowed the separation of PCR amplicons of the same length according to their sequence variations. When the rDNA ITS region was digested with six restriction enzymes, 20 strains were classified into five RFLP haplotypes. The range of similarity between the 20 strains by PCR-RFLP was 42.3-100%. Based on the results of DGGE aud PCR-RFLP, the 20 strains were divided into four groups, R. oryzae, R. stolonifer, R. microsporus and R. homothallicus. Further study of R. homothallicus is required.

• Asian-Australasian Journal of Animal Sciences
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• 제27권10호
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• pp.1493-1498
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• 2014

Epigenetic factors, such as DNA methylation status, may regulate adipogenesis and lipogenesis, thus affecting intramuscular fat (IMF) deposition in longissimus dorsi muscle (LM) of beef cattle. In Korean cattle steers, the LM consists mainly of muscle tissue. However, the LM tissue also contains IMF. We compared the gene expression levels between the IMF and muscle portions of the LM after tissue separation. Real-time polymerase chain reaction analysis showed that the mRNA levels of both adipogenic peroxisome proliferator-activated receptor gamma isoform 1 (PPARG1) and lipogenic fatty acid binding protein 4 (FABP4) were higher (p<0.01) in the IMF than in the muscle portion of the LM. We determined DNA methylation levels of regulatory regions of the PPARG1 and FABP4 genes by pyrosequencing of genomic DNA. DNA methylation levels of two of three CpG sites in the PPARG1 gene promoter region were lower (p<0.05) in the IMF than in the muscle portion of the LM. DNA methylation levels of all five CpG sites from the FABP4 gene promoter region were also lower (p<0.001) in the IMF than in the muscle portion. Thus, mRNA levels of both PPARG1 and FABP4 genes were inversely correlated with DNA methylation levels in regulatory regions of CpG sites of the corresponding gene. Our findings suggest that DNA methylation status regulates tissue-specific expression of adipogenic and lipogenic genes in the IMF and muscle portions of LM tissue in Korean cattle.

• 미생물학회지
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• 제45권2호
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• pp.170-176
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• 2009

제주도에 서식하는 2종의 해양 해면, Dictyonella sp.와 Spirastrella abata의 공생세균 군집구조를 16S rDNA-DGGE(denaturing gradient gel electrophoresis) 방법에 의해 분석하였다. 해면으로부터 total genomic DNA를 추출하여 GC clamp가 추가된 세균에 특이적인 341f primer와 518r primer를 이용하여 16S rRNA gene의 V3 부위를 증폭한 후 DGGE 전기 영동하고 재증폭하여 염기서열을 분석하였다. 그 결과 Dictyonella sp.에서 8개, Spirastrella abata에서 7개의 band를 확인할 수 있었다. 공통된 주요 band가 없는 패턴을 나타내었으며, DGGE band로부터 DNA를 추출하여 부분 염기서열을 분석한 결과, NCBI에 등록된 서열들과 93%~98%의 유사도를 나타내었다. Dictyonella sp.의 주요 해면 공생세균은 uncultured Gammaproteobacteria, Spirastrella abata의 경우 uncultured Alphaproteobacteria, Firmicutes에 각각 포함되어 해면 종에 따른 숙주 특이적 분포를 보이는 것으로 나타났다.

• 한국수산과학회지
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• 제40권6호
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• pp.356-361
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• 2007

Recently, the development of various culture-independent identification techniques for environmental microbes has greatly enhanced our knowledge of microbial diversity. In particular, denaturing gradient gel electrophoresis (DGGE) of 16S rDNA fragments, generated using the polymerase chain reaction (PCR) is frequently used to examine the diversity of environmental bacterial populations. This method consists of direct extraction of the environmental DNA, amplification of the 200-600 bp 16S rDNA fragments with universal primers, and separation of the fragments according to their melting point on a denaturing gradient gel. In this study, we investigated the seaside microbial community in coastal areas of Busan, Korea, using culture-independent techniques. First, marine genomic DNA was extracted from seawater samples collected at Songjeong, Gwangahn, and Songdo Beaches. Then, PCR was used to amplify the bacterial 16S rDNA using universal primers, and DGGE was used to separate the amplified 500 bp 16S rDNA fragments. Finally, the tested 16S rDNA genes were further analyzed by sequencing. Based on these experiments, we found that DGGE analysis clearly showed variation among the regional groups. It can be used to monitor rapid changes in the bacterial diversity of various environments. In addition, the sequence analysis indicated the existence of many unculturable bacteria, in addition to Arcobacter, Pseudoaltermonas, and Vibrio species.