• Title/Summary/Keyword: DNA technology

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Inhibition of Topoisomerase-mediated DNA Cleavage by Lycoperdon perlatum (말불버섯 추출물의 Topoisomerase 저해 효과)

  • Park, Mi-Jung;Cho, Kang-Jin;Kim, Jung-Bong;Kim, Dong-Hern;Kim, Yang-Sub;Seok, Soon-Ja;Kim, Sun-Yeou;Hwang, Young-Soo
    • Korean Journal of Food Science and Technology
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    • v.29 no.5
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    • pp.1057-1062
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    • 1997
  • In the course of searching for anticancer agents from 32 mushrooms, it was found that methanol extract of Lycoperdon perlatum showed inhibitory effect on topoisomerase II-mediated DNA cleavage. This active methanol extract was sequentially fractionated with hexane, chloroform, n-buthanol and water. Among the solvent-fractionated extracts, $1\;{\mu}g/mL$ hexane fraction of L. perlatum inhibited on topoisomerase II-mediated DNA cleavage. The effect of hexane fraction of L. perlatum was dose- and reaction time-dependent. The hexane fraction of L. perlatum was found to have inhibitory activity on relaxation assay of DNA topoisomerase I. The hexane fraction of cultured L. perlatum, however, had no inhibitory effect on either type of topoisomerase.

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Molecular Cloning of the Myosin Light Chain-2 cDNA of Gryllotalpa orientalis

  • Cha, So Young;Hwang, Jae Sam;Kim, Iksoo
    • International Journal of Industrial Entomology and Biomaterials
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    • v.9 no.1
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    • pp.127-130
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    • 2004
  • We describe here the cloning and characterization of a cDNA encoding a putative myosin light chain-2 (MLC-2) from the mole cricket, Gryllotalpa orientalis. The G. orientalis MLC-2 cDNA sequences comprised of 615 bp with 205 amino acid residues with a calculated molecular weight of approximately 23 kDa. The deduced protein sequence of G. orientalis MLC-2 cDNA showed 64% and 54% identity to Drosophila melanogaster MLC-2 and D. yakuba MLC-2, respectively. Northern blot analysis confirmed the muscle-specific expression of G. orientalis MLC-2.

Development of DNA Chip Microarrayer

  • Yoon, Sung-Ho;Choi, Jong-Gil;Lee, Sang-Yup
    • Journal of Microbiology and Biotechnology
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    • v.10 no.1
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    • pp.21-26
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    • 2000
  • A microarrayer system was developed mainly for manufacturing DNA chips. The 3-axis robot was designed to automatically collect samples from 96-or 384-well microtiter plates using up to 16 simultaneously moving pens and to deposit them on a surface-modified slide glass. This is followed by a wash/dry operation in a clean station. The cycle is repeated with a new set of samples, This system can deposit cDNA or oligonucleotides with spot intervals of $150{\;}\mu\textrm{m}$ and the spot size of $80\mu\textrm{m}$, thus allowing a high density DNA chip containing about 5,000 spots per $\textrm{cm}^2$. The entire procedure is controlled by the Visual C++ program that was written in our laboratory by using a personal computer with Pentium 100 CPU.

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A Partial Nucleotide Sequence of Chitin Synthase (CHS) Gene from Rice Blast Fungus, Pyricularia oryzae and Its Cloning

  • Hwang, Cher-Won;Park, In-Cheol;Yeh, Wan-Hae;Takagi, Masamchi;Ryu, Jin-Chang
    • Journal of Microbiology and Biotechnology
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    • v.7 no.2
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    • pp.157-159
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    • 1997
  • A 340-bp chitin synthase gene(CHS) fragment was cloned from the genomic DNA of Pyricularia oryzae using a PCR process with two primer DNAs corresponding to highly conserved sequences within fungal CHS genes. The entire DNA nucleotide sequences of the cloned DNA fragment were determined and analyzed. The amino acid sequences deduced from the nucleotide sequence of the amplified DNA fragment showed 86% homology to that of the Aspergillus fumigatus CHSE gene (9). Using this PCR-amplified DNA, about 2.3 kb of including the PCR fragment of CHSE gene was cloned from genomic library.

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Shelterin Proteins and Cancer

  • Patel, Trupti NV;Vasan, Richa;Gupta, Divanshu;Patel, Jay;Trivedi, Manjari
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.8
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    • pp.3085-3090
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    • 2015
  • The telomeric end structures of the DNA are known to contain tandem repeats of TTAGGG sequence bound with specialised protein complex called the "shelterin complex". It comprises six proteins, namely TRF1, TRF2, TIN2, POT1, TPP1 and RAP1. All of these assemble together to form a complex with double strand and single strand DNA repeats at the telomere. Such an association contributes to telomere stability and its protection from undesirable DNA damage control-specific responses. However, any alteration in the structure and function of any of these proteins may lead to undesirable DNA damage responses and thus cellular senescence and death. In our review, we throw light on how mutations in the proteins belonging to the shelterin complex may lead to various malfunctions and ultimately have a role in tumorigenesis and cancer progression.

Comparison of DNA isolation methods for detection of foodborne pathogens by real-time PCR from foods (식품으로부터 식중독 세균 검출을 위한 Real-time PCR에 적합한 DNA 추출 방법 비교)

  • Koo, Eun-Jeong;Kim, Dongho;Oh, Se-Wook
    • Korean Journal of Food Science and Technology
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    • v.48 no.4
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    • pp.335-340
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    • 2016
  • This study was conducted to find out the most suitable DNA isolation methods for PCR detection of foodborne pathogens. Four DNA isolation methods including Universal Genomic DNA Extraction Kit (TaKaRa), PrepMan Ultra (Applied Biosystems), boiling method and alkaline lysis method (w/PEG) were tested and compared. The Universal Genomic DNA Extraction kit (TaKaRa) was considered as the more efficient isolation method for Escherichia coli O157:H7 and Staphylococcus aureus in lettuce, fish and beef. Meanwhile to detect the foodborne pathogens directly from foods without enrichment, the four different buffers such as double-distilled water, saline, glycine-saline, glycine-saline with Tween-20 and beef extract were also evaluated. As a result, saline was more suitable buffer for E. coli O157:H7. And double-distilled water was more suitable buffer than saline for S. aureus, respectively

Social Issues Arising from the Establishment of a National DNA Database (신원확인 유전자정보은행 설립을 둘러싼 쟁점 연구)

  • Kim Byoung-Soo
    • Journal of Science and Technology Studies
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    • v.3 no.2 s.6
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    • pp.83-104
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    • 2003
  • The use of DNA in identification is growing. The criminal DNA databases are in operation in some countries including the UK, Austria, Germany, and US. The militaries and law enforcement agencies in these countries have used the DNA profile. In Korea, DNA identification has been used in determining paternity and in criminal cases since the middle 1990's, and in recent years law enforcement agencies are promoting a national DNA database for identification. The DNA database threatens our civil liberties because of its potential to be used as an instrument of surveillance. Expanding the database puts increasing numbers of people on a 'list of suspects'. Nevertheless, there is little social concern about using DNA database for identification. This paper reviews social issues related to the establishment of DNA database and investigates the features of DNA profile and DNA Database establishment project promoted law enforcement agencies.

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Dynamic Transcriptome, DNA Methylome, and DNA Hydroxymethylome Networks During T-Cell Lineage Commitment

  • Yoon, Byoung-Ha;Kim, Mirang;Kim, Min-Hyeok;Kim, Hee-Jin;Kim, Jeong-Hwan;Kim, Jong Hwan;Kim, Jina;Kim, Yong Sung;Lee, Daeyoup;Kang, Suk-Jo;Kim, Seon-Young
    • Molecules and Cells
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    • v.41 no.11
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    • pp.953-963
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    • 2018
  • The stepwise development of T cells from a multipotent precursor is guided by diverse mechanisms, including interactions among lineage-specific transcription factors (TFs) and epigenetic changes, such as DNA methylation and hydroxymethylation, which play crucial roles in mammalian development and lineage commitment. To elucidate the transcriptional networks and epigenetic mechanisms underlying T-cell lineage commitment, we investigated genome-wide changes in gene expression, DNA methylation and hydroxymethylation among populations representing five successive stages of T-cell development (DN3, DN4, DP, $CD4^+$, and $CD8^+$) by performing RNA-seq, MBD-seq and hMeDIP-seq, respectively. The most significant changes in the transcriptomes and epigenomes occurred during the DN4 to DP transition. During the DP stage, many genes involved in chromatin modification were up-regulated and exhibited dramatic changes in DNA hydroxymethylation. We also observed 436 alternative splicing events, and approximately 57% (252) of these events occurred during the DP stage. Many stage-specific, differentially methylated regions were observed near the stage-specific, differentially expressed genes. The dynamic changes in DNA methylation and hydroxymethylation were associated with the recruitment of stage-specific TFs. We elucidated interactive networks comprising TFs, chromatin modifiers, and DNA methylation and hope that this study provides a framework for the understanding of the molecular networks underlying T-cell lineage commitment.

Construction of Ovine Customer cDNA Chip and Analysis of Gene Expression Patterns in the Muscle and Fat Tissues of Native Korean Cattle (cDNA microarray를 이용하여 한우의 근육과 지방조직의 유전자 발현 패턴 분석 및 bovine customer cDNA chip 구성 연구)

  • Han, Kyung Ho;Choi, Eun Young;Hong, Yeon-Hee;Kim, Jae Yeong;Choi, In Soon;Lee, Sang-Suk;Choi, Yun Jaie;Cho, Kwang Keun
    • Journal of Life Science
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    • v.25 no.4
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    • pp.376-384
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    • 2015
  • To investigate the molecular events of controlling intramuscular fat (or marbling), which is an important factor in the evaluation of beef quality, we performed cDNA microarray analyses using the longissimus dorsi muscle and back fat tissues. For this study, we constructed normalized cDNA libraries: fat tissues in native Korean cattle (displaying 1,211 specific genes), and muscle tissues in native Korean cattle (displaying 1,346 specific genes). A bovine cDNA chip was constructed with 1,680 specific genes, consisting of 760 genes from muscle tissues and 920 genes from fat tissues. The microarray analysis in this experiment showed a number of differentially expressed genes, which compared the longissimus dorsi muscle (Cy5) with back fat tissue (Cy3). Among many specific differentially expressed genes, 12-lipoxygenase (oxidizing esterified fatty acids) and prostaglandin D synthase (differentiation of fibroblasts to adipocytes) are the key candidate enzymes that should be involved in controlling the accumulation of intramuscular fat. In this study, differentially and commonly expressed genes in the muscle and fat tissues of native Korean cattle were found in large numbers, using the hybridization assay. The expression levels of the selected genes were confirmed by semi-quantitative RT-PCR, and the results were similar to those of the cDNA microarray.

Food Fraud Monitoring of Raw Materials for Commercial Seafood Products Using DNA Barcode Information (DNA Barcode를 이용한 수산가공품 원재료 진위판별)

  • Park, Eun-Ji;Kang, Ju-Yeong;Lee, Han-Cheol;Park, Min-Ji;Yang, Ji-Young;Shin, Ji-Young;Kim, Gun-Do;Kim, Jong-Oh;Seo, Yong-Bae;Kim, Jung-Beom
    • Journal of Food Hygiene and Safety
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    • v.36 no.4
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    • pp.331-341
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    • 2021
  • DNA barcode sequences of commercial seafood products, which are difficult to morphologically discriminate, were analyzed to determine cases of food fraud. The gene sequences were analyzed by amplifying the COX I (cytochrome C oxidase subunit I) gene region of mitochondrial DNA, which is mainly used for species identification. The DNA barcode sequences were compared with the gene sequence of each fish registered in the US National Center for Biotechnology. A total of 46 processed seafood products (12 Pagrus majo, 4 Oplegnathus fasciatus, 7 Dentex tumifrons, 2 Acanthopagrus schlegelii, 7 Oreochromis niloticus, 6 Branchiostegus japonicus, 8 Branchiostegus albus) were investigated. Having DNA sequence identity of more than 97% was judged as the same species. As a result of this study, no cases of forgery and alteration were detected. However, some disparities in the commercial names used in local markets and the standard names given in the Korea Food Code were found, which may cause confusion for consumers. It is therefore suggested that the standard name or scientific name be displayed on seafood product labels.