• 대한수의학회지
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• 제55권3호
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• pp.163-167
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• 2015

Canine parvovirus (CPV) is a major diarrhea-causing agent in puppies. Since CPV type 2 (CPV-2) emerged in 1978, new antigenic variants including CPV-2a, CPV-2b, and CPV-2c have been identified in many countries. Two puppies died suddenly at a veterinary clinic in Gyeonggi province, South Korea. Two viruses were isolated in A72 cells, confirmed as CPV strains based on a CPV rapid kit and an indirect fluorescence test and designated QIACP1403 and QIACP1404. The nucleotide sequences of complete VP2 genes of QIACP1403 and QIACP1404 were determined, and the corresponding amino acid sequences were deduced. Molecular analyses revealed that the QIACP1403 and QIACP1404 isolates were type CPV-2b. Several mutated amino acids were detected on VP2 gene residues of the two isolates. Phylogenetic analyses showed that the two isolates were most closely related to strain CPV-BM11, which was isolated from Chinese dogs in 2011. Our results suggest that these isolates may be a candidate for a vaccine to prevent CPV infection in dogs after conducting passages of the isolates in an in vitro culture system.

• 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.

• 대한의생명과학회지
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• 제15권4호
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• pp.295-300
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• 2009

Tuberculosis caused by Mycobacterium tuberculosis (MTB) still remains to be the most dreadful infectious disease affecting almost every country. In the present study, we developed a simple and rapid but accurate and sensitive assay method for detecting MTB using microplate hybridization assay. For this, a selective region of the rpoB gene was used to design PCR primers, and MTB and Mycobacterium genus-specific probe molecules. The specificity of the assay was confirmed using fifteen different mycobacterial reference strains and twelve different non-mycobacterial reference strains, and the sensitivity was determined to be 100 fg using genomic DNA of MTB reference strain, H37Rv. Subsequently, a total of 62 sputum samples with diverse smear scores and culture positive results were used to evaluate the kit performance. In brief, the specificity and the sensitivity of the assay were 100% and 98.4%, respectively.

• Communications for Statistical Applications and Methods
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• 제16권6호
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• pp.1037-1046
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• 2009

Copy number variations(CNVs) are known as one of the most important factors in susceptibility to genetic disorders because they affect expression levels of genes. In previous studies, pyrosequencing, mini-sequencing real-time polymerase chain reaction(PCR), invader assays and other techniques have been used to detect CNVs. However, the higher the copy number in a genome, the more difficult it is to resolve the copies, so a more accurate method for measuring CNVs and assigning genotype is needed. PCR followed by a quantitative oligonucleotide ligation assay(qOLA) was developed for quantifying CNVs. The aim of this study was to compare the two methods for detecting and quantifying the CNVs of duplicated gene: the published pyrosequencing assay(pyro_CNV) and the newly developed qOLA_CNV. The accuracy and precision of the assay were evaluated for porcine KIT, which was selected as a model locus. Overall, the root mean squares(RMSs) of bias and standard deviation of qOLA_CNV were 2.09 and 0.45, respectively. These values are less than half of those of pyro CNV.

• 한국동물위생학회지
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• 제26권2호
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• pp.105-111
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• 2003

From February 2000 to December 2001, A total of 1,785 samples was taken from beef and pork carcasses in Seoul. Seven(0.69%) Listeria spp. were isolated from the 1,014 of beef carcasses, and five(0.65%) were isolated from the 771 of pork carcasses. The isolates were identified L monocytogenes by API listeria, and VIDAS LMO kit, serological test and PCR assay were preformed. A total 12 strains of L monocytogenes were isolated form samples tested and all of the strains were classified into serotype 1. PCR primers are selected to amplify a 520-base pair(bp) DNA fragment from the listeolysin O gene(hlyA) of Listeria monocytogenes. A 520-bp product was detected in PCR with DNA from L monocytogenes, but not from the other Listeria species tested.

• Food Science and Biotechnology
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• 제18권2호
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• pp.384-389
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• 2009

To isolate lactic acid bacteria having high exopolysaccharides (EPS) production ability, 50 strains were initially isolated from the sourdough. Twenty-one strains formed highly mucoid colonies on the sucrose agar medium, which are indicative of active EPS synthesis. DU-07, DU-10, DU-12, DU-19, and DU-21 produced $11.51{\pm}0.167$, $13.09{\pm}0.193$, $12.72{\pm}0.108$, $11.61{\pm}0.284$, and $13.32{\pm}0.094\;g/L$ EPS, respectively, in MRS medium. The isolated strains, DU-10, DU-12, and DU-21, were identified as Enterococcus flavescens, Enterococcus faecium, and Lactobacillus amylovorus, respectively, by using API 50CHL kit and determining partial sequences of their 16S rDNA. Especially, L. amylovorus DU-21 showed the highest production of EPS, as well as the highest inhibitory activities against pathogenic (p<0.05). Interestingly, the L. amylovorus DU-21 seem to be endemic to sourdough fermentations, as they have not been isolated from other environments.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2018년도 춘계학술발표회
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• pp.80-80
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• 2018

Chlorogenic acid is a phenolic compound found in Cudrania tricuspidata fruits. In the present study, the effect of chlorogenic acid on the inhibition of adipogenesis in 3T3-L1 adipocytes was investigated. Cells were stained with Oil red O reagent to detect lipid droplets in adipocytes. The 3T3-L1 cells were lysed and measured for intracellular triglyceride and adipokine by ELISA kit. The protein expression of adipogenesis-related gene was evaluated by Western blot analysis. Chlorogenic suppressed lipid droplet and intracellular triglyceride accumulation in a concentration manner and also decreased secretion of adipokines such as leptin and adiponectin, compared with fully differentiated adipocytes. Treatment of 3T3-L1 cells with chlorogenic acid reduced the protein levels of peroxisome proliferator-activated receptor gamma ($PPAR{\gamma}$) and, CCAAT/enhancer binding proteins alpha ($C/EBP{\alpha}$). This indicates that chlrogenic acid was effective as an anti-obesity agent by repressing the differentiation of 3T3-L1 into adipocytes and inhibiting triglyceridef formation in adipocyte and that it exerts its role mainly through the significant down-regulation of $PPAR{\gamma}$ and $C/EBP{\alpha}$.

• Journal of Species Research
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• 제12권3호
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• pp.197-202
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• 2023

The purpose of this study was to isolate and identify wild yeasts from the soil collected in Gwangju and Pocheon City, Gyeonggi Province, Republic of Korea. Among 10 strains, six strains were already reported, but four strains were unrecorded in Republic of Korea. To identify wild yeast strains, pairwise sequence comparisons of the D1/D2 region of the 26S rRNA gene sequence were performed using Basic Local Alignment Search Tool (BLAST). The cell morphologies were observed by phase contrast microscope and assimilation tests were carried out using API 20C AUX kit. The 10 strains were assigned to the phyla Basidiomycota (8 strains) and Ascomycota (2strains). The unrecorded four yeast strains, NH33, NH19, NH20, and YP416, belong to the phylum Basidiomycota and the genera Buckleyzyma, Leucosporidium, Holtermanniales, and Mrakia, respectively. All strains had oval-shaped and polar budding cells. In this research, the morphological and biochemical properties of four unreported yeast species were characterized intensively, which were not officially reported in Korea.

• 생명과학회지
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• 제28권7호
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• pp.835-840
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• 2018

본 연구는 시토크롬 P450 단백질을 암호화하는 애기장대 유래의 AtCYP78A7을 과발현하는 형질전환 식물체로부터 AtCYP78A7 단백질을 특이적으로 인식하는 단일큰론 항체의 제조와 그 항체를 AtCYP78A7 단백질과 접촉시켜 항원-항체 복합체 형성을 검출함으로써 AtCYP78A7 단백질을 효소면역학적(ELISA) 방법으로 검출하는 진단 키트를 개발하기 위하여 수행하였다. 재조합한 GST-AtCYP78A7 단백질을 항원으로 사용하여 단일클론 항체를 분비하는 융합세포주를 제조한 후 비오틴화 및 페어링 테스트를 통해 포획항체와 검출항체를 선정하였으며, GST-AtCYP78A7 정제 단백질을 기준으로 일품벼, 화영벼, AtCYP78A7 과발현 벼(10B-5, 18A-4)의 용해물을 검출항원으로 사용하여 product test를 진행하였다. 그 결과 AtCYP78A7 단백질에 특이적으로 결합하는 4개의 단클론 항체(mAb 6A7, mAb 4C2, mAb 11H6, mAb 7E8)를 생산하였고, 포획항체 mAb 4C2와 검출항체 mAb 7E8-biotin의 조합으로 ELISA 키트를 개발하였다. 개발된 ELISA 키트를 이용한 벼 시료의 분석 결과 AtCYP78A7 과발현 벼는 전체 단백질 대비 AtCYP78A7 단백질의 비율이 0.1% 이상인 양성으로, 일품벼와 화영벼는 0.1% 미만인 음성으로 나타나 키트를 이용한 AtCYP78A7 단백질의 검출이 가능하였으며, 따라서 본 키트는 향후 AtCYP78A7를 과발현하는 형질전환 작물을 대상으로 하는 환경 모니터링 또는 인체 위해성 평가에 유용하게 활용될 수 있을 것으로 사료된다.

• 분석과학
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• 제29권2호
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• pp.79-84
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• 2016

최근 손안에 들어올 정도로 크기가 작아 범죄현장 등에서 사용이 가능하며, 다른 일반적인 장비들에 비해 가격이 1/10이하로 저렴하여 누구나 사용할 수 있는 MiniPCR^TM mini8 Thermal Cycler (Amplyus, Cambridge, MA, USA)가 개발되었다. 본 연구에서는 DNA감식에 일반적으로 사용되고 네 가지 종류의 상염색체 STR 다중증폭 키트들과 한 종류의 Y 염색체 STR 증폭키트, 그리고 미토콘드리아 DNA HV1/HV2의 염기서열 분석법을 사용하여 MiniPCR^TM mini8 Thermal Cycler의 성능을 Applied Biosystems사의 GeneAmp^® PCR system 9700와 비교하였다. STR 다중증폭키트 키트들의 민감도와 증폭 불균형 정도를 비교한 결과 두 PCR 장비에서 큰 차이가 없었으며, 미토콘드리아 DNA HV1/HV2의 염기서열 분석 결과도 동등하였다. MiniPCR^TM mini8 Thermal Cycler는 DNA 감식 실험실은 물론이고, 가격이 저렴해 학교와 개인이 간편하게 사용할 수 있으며, 휴대가 간편해 차량이나 야외에서 활용 될 수 있을 것으로 기대된다.