• 대한전기학회:학술대회논문집
• /
• 대한전기학회 2006년도 제37회 하계학술대회 논문집 C
• /
• pp.1324-1326
• /
• 2006

In this study, an integrated microelectrode array was fabricated on glass slide using microfabrication technology. Probe DNAs consisting of mercaptohexyl moiety at their 5-end were spotted on the gold electrode using micropipette or DNA arrayer utilizing the affinity between gold and sulfur. Cyclic voltammetry in 5mM ferricyanide/ferrocyanide solution at 100 mV/s confirmed the immobilization of probe DNA on the gold electrodes. When several DNAs were detected electrochemically, there was a difference between target DNA and control DNA in the anodic peak current values. It was derived from specific binding of Hoechst 33258 to the double stranded DNA due to hybridization of target DNA. It suggested that this DNA chip could recognize the sequence specific genes. It suggested that multichannel electrochemical DNA microarray is useful to develop a portable device for clinical gene diagnostic system.

• 대한전기학회:학술대회논문집
• /
• 대한전기학회 2006년도 제37회 하계학술대회 논문집 C
• /
• pp.1319-1321
• /
• 2006

This research aims to develop the multiple channel electrochemical DNA chip that has the above characteristic and be able to solve the problems. At first, we fabricated a high integration type DNA chip array by lithography technology. It is able to detect a plural genes electrochemically after immobilization of a plural probe DNA and hybridization of non-labeling target DNA on the electrodes simultaneously. It suggested that this DNA chip could recognize the sequence specific genes. It suggested that multichannel electrochemical DNA microarray is useful to develop a portable device for clinical gene diagnostic system.

• Journal of Animal Science and Technology
• /
• 제44권1호
• /
• pp.13-22
• /
• 2002

한우의 성장단계 특이발현 유전자를 탐색하기 위하여, 본 연구에서는 유전자의 발현 유무 및 발현정도의 차이를 나타내는 유전자를 분리하는데 있어 가장 강력한 수단으로 알려진 subtractive cDNA hybridization 기법을 이용하여 한우 등심조직으로부터 12개월령 및 24개월령 특이적인 subtractive cDNA library를 제작하였다. 성장단계 특이적인 유전자를 탐색하기 위하여, 6, 12 및 24개월령 cDNA를 사용하여 reverse northern blot 분석을 실시하였으며, 6개월령 cDNA probe에 대하여 특이적인 signal을 나타낸 3개의 clone은 EPV 20, Ca2+ ATPase, 및 TCTP 유전자와 유사성을 나타내었다. 12개월령 cDNA probe에 대하여 특이적인 signal을 나타낸 9개의 cDNA clone은 각각 VCP, HSP 70, aldolase A, MSSK1, GM-2 activator protein, ryanodine receptor, acidic ribosomal phosphoprotein p1, ADP/ATP translocase T1 및 UCP 2 유전자와 높은 homology를 가지고 있었다. 또한 2개의 clone이 각각 12개월령 및 24개월령 cDNA probe에 대하여 특이적인 signal을 나타내었는데, 12개월령 cDNA probe에 대해서만 signal을 나타낸 clone은 ferrochelatase 유전자와 유사하였으며, 24개월령 probe에 대해서만 signal을 나타낸 clone은 ADRP 유전자와 유사하였다. 이상에서와 같이, 본 연구에서 제작한 성장단계 특이적인 subtractive cDNA library를 분석하여 14종의 유전자를 한우 성장단계 특이 발현 후보 유전자로 선정하여 염기서열을 분석하였으며, 이외에도 성장단계에 있어 특이적으로 발현될 것으로 추정되는 cDNA 클론의 염기서열을 분석하였다.

• 한국식품과학회지
• /
• 제48권2호
• /
• pp.128-132
• /
• 2016

PEG를 이용하여 WGA 수행 시 DNA 증폭 효율을 높이고 이를 식중독 세균의 DNA 증폭 및 검출에 적용하고자 하였다. 등온 증폭 반응인 WGA에 여러 종류의 PEG를 첨가하여 증폭한 결과, 1.5% 농도의 PEG 4,000을 첨가하는 것이 가장 효율이 높음을 알 수 있었다. 증폭 정도를 정량적으로 파악하기 위하여 3종의 식중독 세균 DNA를 이용하여 WGA를 수행하였으며 real-time PCR로 정량분석하였다. S. Typhimurium, L. monocytogenes, V. parahaemolyticus의 경우에 WGA를 하지 않은 DNA에 비하여 각각 7,777.01배, 9,981.22배, 1,239.03배 정도로 DNA의 양이 증폭되는 것을 확인하였다. 또한 PEG를 첨가함으로써 18배에서 40배의 핵산 증폭 효과가 더 있음을 알 수 있었다. 따라서 식품에 미량의 농도로 존재하는 식중독 세균은 PEG가 첨가된 WGA 반응을 통하여 검출 가능성을 높일 수 있음을 알 수 있었다.

• ALGAE
• /
• 제35권3호
• /
• pp.293-301
• /
• 2020

The applications of DNA barcoding have a wide range of uses, such as in taxonomic studies to help elucidate cryptic species and phylogenetic relationships and analyzing environmental samples for biodiversity monitoring and conservation assessments of species. After obtaining the DNA barcode sequences, sequence similarity-based homology analysis is commonly used. This means that the obtained barcode sequences are compared to the DNA barcode reference databases. This bioinformatic analysis necessarily implies that the overall quantity and quality of the reference databases must be stringently monitored to not have an adverse impact on the accuracy of species identification. With the development of next-generation sequencing techniques, a noticeably large number of DNA barcode sequences have been produced and are stored in online databases, but their degree of validity, accuracy, and reliability have not been extensively investigated. In this study, we investigated the extent to which the amount and types of erroneous barcode sequences were deposited in publicly accessible databases. Over 4.1 million sequences were investigated in three largescale DNA barcode databases (NCBI GenBank, Barcode of Life Data System [BOLD], and Protist Ribosomal Reference database [PR2]) for four major DNA barcodes (cytochrome c oxidase subunit 1 [COI], internal transcribed spacer [ITS], ribulose bisphosphate carboxylase large chain [rbcL], and 18S ribosomal RNA [18S rRNA]); approximately 2% of erroneous barcode sequences were found and their taxonomic distributions were uneven. Consequently, our present findings provide compelling evidence of data quality problems along with insufficient and unreliable annotation of taxonomic data in DNA barcode databases. Therefore, we suggest that if ambiguous taxa are presented during barcoding analysis, further validation with other DNA barcode loci or morphological characters should be mandated.

• 분석과학
• /
• 제11권2호
• /
• pp.1013-1024
• /
• 1998

• 한국생물공학회소식지
• /
• 제12권2호
• /
• pp.20-25
• /
• 2005

• 한국버섯학회지
• /
• 제11권3호
• /
• pp.171-176
• /
• 2013

큰느타리버섯의 저온성적응성 형질에 관련된 SCAR marker를 개발하기 위하여 저온성 계통의 8균주와 대조구 8균주의 genomic DNA를 30 ug/ml의 농도로 bulking한 것을 주형 DNA로 사용하고 operon 사의 OPA(20개), OPB(20개), OPL(20개), OPP(20개), OPR(20개), OPS(20개) 등 총 120개 primer를 random primer(10 mer)로 사용하여 RAPD를 수행하였으며 이중에서 OP-S3 primer를 사용한 PCR 산물들이 대조구와 가장 뚜렷한 차이를 나타내었다. OP-S3 primer를 이용한 RAPD 결과, 약 480 bp 부근에서 저온성 계통에 특이적인 DNA band가 관찰되었으며 이 DNA band의 염기서열을 근거로 SCAR marker로 사용할 specific primer인 OP-S3-1-F와 OP-S3-1-R를 디자인하였다. SCAR marker OP-S3-1 primer를 이용하여 PCR을 수행한 결과에서는 저온성 계통에서만 480 bp 부근에서 대조구와 구별되는 DNA band를 확인할 수 있었으며 random primer인 OP-S3 primer를 이용하여 PCR을 수행했을 때보다 재현성이 높고 진한 DNA band를 확인할 수 있었다.

• 한국환경성돌연변이발암원학회지
• /
• 제22권3호
• /
• pp.164-168
• /
• 2002

Phthalate analogues are a plasticizer and solvent used in industry and were reported to be a potential carcinogen classified in the category of suspected endocrine disruptors. Most common human exposure to these compounds may occur with contaminated food. They may migrate into food from plastic wrap or may enter food from general environmental contamination. Since these substances are not limited to the original products, and enter the environment, they have become widespread environmental pollutants, thus leading to a variety of phthalates that possibly threaten the public health. To determine whether seven phthalate analogues i.e. diallyl phthalate, diisodecyl phthalate, di-n-nonyl phthalate, butyl benzyl phthalate, di-n-octyl phthalate, di-tridecyl phthalate, and dibutyl phthalate, can induce DNA strand breakage that is one of the various factors related to the mechanism of carcinogenicity, the comet assay which has been widely used for the detection and measurement of DNA strand breaks, was conducted in L5178Y mouse lymphoma cells. From these results, seven phthalates revealed dose-dependent decrease of cell viability, however, no remarkable cytotoxicity was observed even at high concentration of 100 $\mu\textrm{g}$/$m\ell$ phthalates. And also, the results showed that the induction of DNA strand breaks by seven phthalates was not significantly different from the control in this study.

• Annals of Clinical Neurophysiology
• /
• 제19권1호
• /
• pp.40-45
• /
• 2017

Background: Telbivudine is a nucleoside analogue used for the treatment of chronic hepatitis B, but it often develops mitochondrial toxicity leading to symptomatic myopathy. In this study, three patients with telbivudine induced myopathy were enrolled in order to investigate the nature and pathogenesis of mitochondrial toxicity caused by long-term use of telbivudine. Methods: Clinical features, laboratory findings, muscle pathology, and quantitation of mitochondrial DNA were studied in three patients. Results: Patients presented with progressive muscle weakness with high serum creatine kinase levels. Light microscopic findings of muscle pathology showed ragged red fibers that reacted strongly with succinate dehydrogenase stain, but negative for cytochrome c oxidase activities. Electron microscopy revealed abnormal mitochondrial accumulation with rod shaped inclusions. The quantitative peroxidase chain reaction showed a depletion of mitochondrial DNA in skeletal muscle of the patients. Conclusions: Nucleoside analogues including telbivudine are potent inhibitors of viral DNA polymerases. However, they are not specific for viral DNA and can disturb mitochondrial replication at the same time. All nucleotide analogues should be used with close clinical observation in order to avoid development of mitochondrial myopathy.