• Journal of Applied Biological Chemistry
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• 제48권3호
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• pp.113-119
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• 2005

O-methylation mediated by O-methyltransferases (OMTs) is a common modification in natural product biosynthesis and contributes to diversity of secondary metabolites. OMTs use phenylpropanoids, flavonoids, other phenolics and alkaloids as substrates, and share common domains for S-adenosyl-L-methionine (AdoMet) and substrate binding. We searched Arabiposis genome and found 17 OMTs genes (AtOMTs). AdoMet- and substrate-binding sites were predicted. AdoMet binding domain of AtOMTs is highly conserved, while substrate-binding domain is diverse, indicating use of different substrates. In addition, expressions of six AtOMT genes in response to UV and in different tissues were investigated using real-time quantitative reverse transcriptase-polymerase chain reaction. All the AtOMTs investigated were expressed under normal growth condition and most, except AtOMT10, were induced after UV illumination. AtOMT1 and AtOMT8 were expressed in all the tissues, whereas AtOMT10 showed flower-specific expression. Analysis of these AtOMT gene expressions could provide some clues on AtOMT involvement in the cellular processes.

• Parasites, Hosts and Diseases
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• 제56권4호
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• pp.371-374
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• 2018

A 3-month-old female Maltese puppy was hospitalized with persistent diarrhea in a local veterinary clinic. Blood chemistry and hematology profile were analyzed and fecal smear was examined. Diarrheal stools were examined in a diagnostic laboratory, using multiplex real-time polymerase chain reaction (PCR) against 23 diarrheal pathogens. Sequence analysis was performed using nested PCR amplicon of 18S ribosomal RNA. Coccidian oocysts were identified in the fecal smear. Although multiplex real-time PCR was positive for Cyclospora cayetanensis, the final diagnosis was Cystoisospora ohioensis infection, confirmed by phylogenetic analysis of 18S rRNA. To our knowledge, this the first case report of C. ohioensis in Korea, using microscopic examination and phylogenetic analysis.

• 한국발생생물학회지:발생과생식
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• 제24권3호
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• pp.167-176
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• 2020

The fat pad defined as the epididymal fat is located at the head part of the epididymis. However, another fat mass is present near the caudal epididymis, named tail epididymal fat. The present research was focused to determine the expression of adipocyte-associated molecules in the mouse tail epididymal fat at different postnatal ages, including 2, 5, 8, and 12 months of age. The quantitative real-time PCR analysis showed continuous increases of expression levels of delta like non-canonical Notch ligand 1, leptin, and resistin as postnatally aged. The transcript level of peroxisome proliferator-activated receptor gamma was the highest at 5 months of age, remaining at a steady level until 12 months of age. Expression levels of fatty acid binding protein 4, leptin, and adiponectin were significantly increased until 8 months of age, persisting the level at 12 months of age. The transcript level of fatty acid synthase was significantly increased at 8 months of age, without a further change of the level at 12 months of age. These findings exhibited the expression of adipocyte-associated genes which were also detected at the ordinary epididymal fat pad. However, expression patterns of these genes in the tail epididymal fat are different with those in the distal and proximal epididymal fat, suggesting distinct characteristics and/or functions of the tail epididymal fat.

• Genomics & Informatics
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• 제21권2호
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• pp.24.1-24.7
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• 2023

Assays of clinical diagnosis and species identification using molecular markers are performed according to a quantitative method in consideration of sensitivity, cost, speed, convenience, and specificity. However, typical polymerase chain reaction (PCR) assay is difficult to quantify and have various limitations. In addition, to perform quantitative analysis with the quantitative real-time PCR (qRT-PCR) equipment, a standard curve or normalization using reference genes is essential. Within the last a decade, previous studies have reported that the digital PCR (dPCR) assay, a third-generation PCR, can be applied in various fields by overcoming the shortcomings of typical PCR and qRT-PCR assays. We selected Stilla Naica System (Stilla Technologies), Droplet Digital PCR Technology (Bio-Rad), and Lab on an Array Digital Real-Time PCR analyzer system (OPTOLANE) for comparative analysis among the various droplet digital PCR platforms currently in use commercially. Our previous study discovered a molecular marker that can distinguish Hanwoo species (Korean native cattle) using Hanwoo-specific genomic structural variation. Here, we report the pros and cons of the operation of each dPCR platform from various perspectives using this species identification marker. In conclusion, we hope that this study will help researchers to select suitable dPCR platforms according to their purpose and resources.

• 한국식품저장유통학회지
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• 제30권3호
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• pp.383-394
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• 2023

The ultra-fast quantitative real-time polymerase chain reaction (qPCR) assay was developed and validated to differentiate the morphologically similar ones, Oncorhynchus keta and Oncorhynchus mykiss. Species-specific primers were designed for the COI genes of mtDNA. The species-specific primers designed for O. keta and O. mykiss were selectively amplified by O. keta and O. mykiss DNA, respectively. The sensitivity of O. keta and O. mykiss primers was 1 ng/μL. Quantitative testing showed that the results met the 'Guidelines on Standard Procedures for Preparing Analysis Method such as Food' proposed by the Ministry of Food and Drug Safety. The qPCR method developed and validated in this study for identifying O. keta and O. mykiss has advantages such as speed and field applicability. Therefore, this method is expected to help control forgery and alteration of raw materials in the seafood industry.

• Journal of Dairy Science and Biotechnology
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• 제33권3호
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• pp.197-202
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• 2015

While various foodborne pathogenic bacteria can be detected more rapidly via polymerase chain reaction than via conventional plating methods, it is impossible to distinguish between viable and dead cells in DNA-based assays. Hence, propidium monoazide (PMA) treatment has been introduced to detect living cells. The purpose of this study is to evaluate the applicability of the PMA treatment and real-time qPCR method for the detection of Cronobacter sakazakii and to compare it to that of plate counting. Based on our positive results, we suggest the use of PMA treatment and real-time qPCR for the detection of viable Cronobacter sakazakii in various food sources and an update of the Korean Food Code.

• 보존과학연구
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• 통권32호
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• pp.171-183
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• 2011

본 연구는 충청남도 부여군 규암면 오수리 유적에서 발굴된 인골 4개체를 대상으로 조직학, 분자유전학, 골화학 분석 등 종합적인 연구를 통해 이들의 상관관계를 규명하고자 하였다. 실체현미경을 통해 인골 시료의 골조직 단면 구조를 관찰함으로써 각 시료의 조직학적 보존 상태를 단계별로 구분하였고, 잔존하는 단백질의 보존 상태는 콜라겐을 추출하여 수율을 측정함으로써 평가하였다. 또한 미토콘드리아 cytochrome b 유전자를 이용한 실시간 유전자 증폭법을 이용하여 각각의 인골 시료에 잔존하는 미토콘드리아 DNA의 상대적 보존량 및 복제수를 분석하였으며, 미토콘드리아 과변위부위의 염기서열을 동정하였다. 본 연구 결과 인골 시료의 조직학적 보존정도, 콜라겐 단백질의 잔존량, 미토콘드리아 DNA의 복제수는 상호 긍정적인 연관관계로 나타났다. 이 연구는 출토 인골의 생물 화학적 분석 가능성을 예측하기 위한 특성지표 연구의 중요자료로 활용 될 수 있을 것이다.

• 한국축산식품학회지
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• 제29권6호
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• pp.668-672
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• 2009

This paper describes the differentiation between native Korean cattle (Hanwoo) and Holsteins or imported cattle using the real-time polymerase chain reaction (PCR) by targeting the sequence of the melanocortin 1 receptor (MC1R) gene. A rapid and accurate method was developed to identify Hanwoo by genotyping the DNA extracted from 295 commercial beef samples (obtained from 5 provinces in South Korea) labeled as Hanwoo beef. The results of real-time PCR assays for the proportions of Hanwoo were 84, 85.7, 95, 91.4, and 90% in the areas of Seoul, Joongbu, Youngnam, Honam, and Chungcheong, respectively. Thus, the beef samples from 295 butcher shops, which asserted to only sell Hanwoo, showed that 259 of 295 samples were of the Hanwoo beef gene type (T-type) and 36 of 295 samples were Holsteins of imported dairy cattle gene types (C-type or C/T type). In conclusion, the proportion of Hanwoo beef was 87.8% and the proportion of Holstein or imported dairy cattle meat was 12.2% (C-type: 9.8%, C/T-type: 2.4%). Generally, most consumers can not differentiate imported meat from Hanwoo beef. Therefore, Hanwoo beef and imported dairy cattle meat that is sold in butcher shops should have mandatory identification by using MC1R genotyping based on real-time PCR.

• Journal of Periodontal and Implant Science
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• 제51권1호
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• pp.53-62
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• 2021

Purpose: This study aimed to evaluate the clinical and microbiological efficacy of adjunctive local delivery of minocycline (Periocline^®) in patients receiving supportive periodontal therapy (SPT) after initial treatment. Methods: The participants were 16 men and 8 women (age, 20-65 years) who had at least 15 natural teeth, underwent SPT for more than 1 year due to chronic periodontitis, had 4 or more periodontal pocket sites deeper than 5 mm, and showed >25% gingival bleeding on probing (BoP). They were randomly assigned to the test and control groups. In the test group, mechanical debridement and local antibiotic delivery were performed for all periodontal sulci/pockets; in the control group, mechanical debridement and saline irrigation were performed. In patients who underwent SPT for more than 1 year, clinical and microbiological examinations were performed at baseline and 1 and 3 months after SPT. The clinical examination included an assessment of the periodontal pocket depth, clinical attachment level, plaque index, and BoP. Microbial tests were performed using real-time polymerase chain reaction; the relative ratios of Porphyromonas gingivalis and Fusobacterium nucleatum were determined. Results: Both groups showed significant improvements in clinical parameters at 1 and 3 months from baseline; there were no significant changes between months 1 and 3. Intergroup differences were insignificant. The microbiological analysis revealed no significant differences in P. gingivalis and F. nucleatum ratios across time points. While intergroup differences were insignificant, there was a tendency for the P. gingivalis and F. nucleatum ratios to decrease in the test group. Conclusions: Mechanical debridement in patients receiving maintenance therapy resulted in clinically significant improvement; the effectiveness of additional local delivery of antibiotics was not significant. The ratios of P. gingivalis and F. nucleatum showed a tendency to decrease in the test group, although it was not significant.

• Tuberculosis and Respiratory Diseases
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• 제62권5호
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• pp.406-416
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• 2007

연구배경: 단일염기다형성(Single nucleotide polymorphism, SNP)은 인간의 유전자 서열 1000염기에 1개 빈도로 발견되어 인간은 대략 300만개의 유전자 다형성을 가지고 있다. 이 유전자 다형성의 조합결과로 인간의 개체 간 특성들이 결정되는 것으로 이해되고 있다. 이러한 다형성들의 조합양상에 따라 특이 질환에 대한 유전자 감수성 또한 달라지게 되므로 최근에는 많은 질환들과 유전자 다형성들과의 상관관계를 보는 연구들도 활발하게 진행되고 있다. 이러한 SNP분석은 큰 집단을 대상으로 진행되어 지므로 적은 비용으로 정확하게 그리고 대용량으로 분석할 수 있는 방법이 필요하다. 방 법: 대상 환자 89명의 genomic DNA를 가지고서 promotor상에 위치한 -37과 -524 염기부위에서 유전자 다형성을 보이는 것으로 보고되어져 있는 RRM1(ribonucleotide reductase M1) 유전자를 대상으로 PCR-RFLP(polymerase chain reaction-restriction fragment length polymorphism)와 real-time PCR(RTPCR, TaqMan probe assay)을 동시에 시행한 후 각각의 결과를 비교 분석하였다. 결 과: 대상 DNA 89예 중 -37에서는 2예(2.17%), -524에서는 15예(16.26%)가 서로 다른 양상을 보였다. 결과 차이를 보인 샘플 17예를 대상으로 직접 염기서열 분석을 시행하여 본 결과, 17예 모두 RT-PCR에서 확인되었던 결과와 일치함을 확인할 수 있었다. 추가 샘플 138예를 대상으로 RT-PCR을 2회 연속 실행하여 genotyping을 해 본 결과 98%이상의 높은 일치율을 보였으며, 그중 10예를 무작위로 골라 직접 염기서열 분석을 시행하여 본 결과, 역시 100%일치, 높은 정확도를 보였고 이는 in-tube assay 방식으로 샘플의 오염을 최소화 할 수 있었으며 72 well based system(Corbett Research)을 이용함으로 1회 유전자 증폭반응을 통해 많은 검체를 한 번에 확인할 수 있어 매우 빠른 검사방법 이었다. 결 론: 큰 집단을 대상으로 다량의 SNP를 분석하기 위한 실험 방법으로는 RT-PCR이 신속하면서도 정확한 결과를 얻을 수 있는 방법으로 사료된다.