• Title/Summary/Keyword: DNA Processing

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Intra- and Extra-cellular Mechanisms of Saccharomyces cerevisiae Inactivation by High Voltage Pulsed Electric Fields Treatment (고전압 펄스 전기장에 의한 Saccharomyces cerevisiae의 세포내·외적 사멸 기작 연구)

  • Lee, Sang-Jae;Shin, Jung-Kue
    • Korean Journal of Food Science and Technology
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    • v.47 no.1
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    • pp.87-94
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    • 2015
  • High voltage pulsed electric fields (PEF) treatment is one of the more promising nonthermal technologies to fully or partially replace thermal processing. The objective of this research was to investigate the microbial inactivation mechanisms of PEF treatment in terms of intra- and extracellular changes in the cells. Saccharomyces cerevisae cells treated with PEF showed cellular membrane damage. This resulted in the leakage of UV-absorbing materials and intracelluar ions, which increased with increasing treatment time and electric fields strength. This indicates that PEF treatment causes cell death via membrane damage and physical rupture of cell walls. We further confirmed this by Phloxine B staining, a dye that accumulates in dead cells. Using scanning and transmission electron microscopy, we observed morphological changes as well as disrupted cytoplasmic membranes in PEF treated S. cerevisae cells. In addition, PEF treatment led to damaged chromosomal DNA in S. cerevisiae.

Isolation and Characterization of Dehydrin 1 (Dhn1) gene from Codonopsis lanceolata (더덕의 주근에서 유래한 Dehydrin 1 (Dhn1) 유전자의 분리 및 분석)

  • Lee, Kang;Yang, Duk-Chun
    • Korean Journal of Plant Resources
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    • v.16 no.3
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    • pp.238-244
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    • 2003
  • During the life cycle, plants have to suffer from various environmental stresses. A common element in response to many environmental stresses is cellular dehydration. Dehydrins are a family of proteins commonly induced by environmental stresses associated with low temperature or dehydration and during seed maturation drying. For the study in the defense mechanism against various stresses, a cDNA clone encoding a dehydrin gene was isolated from a cDNA library prepared from tab root mRNAs of Codonopsis lanceolata. The cDNA, designated ClDhn1, is 893 nucleotides long and has an open reading frame of 480 bp with a deduced amino acid sequence of 159 residues. The ClDhn1 amino acid sequence is highly hydrophilic and possesses two conserved repeats of characterized lysine­rich K­segment (KIKEKLPG), and a 7­serine residue stretch prior to the first lysine­rich repeat that is common to many dehydrins. The DEYGNP conserved motif is, however, modified in the sequence of ClDhn1 gene. The deduced amino acid sequence of ClDhn1 was compared with other plant dehydrinls and showed high homology with Solanum commersonii

Evaluation on the Applicability of Comet Assay for the Identification of Gamma-irradiated Frozen Beef Cut, Ground Beef and Tteokgalbi (감마선처리 냉동 우육, 분쇄우육 및 떡갈비의 판별을 위한 Comet assay의 적용성 평가)

  • Jong-Heum Park;Jae-Kyung Kim;Beom-Seok Song
    • Journal of Radiation Industry
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    • v.17 no.3
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    • pp.257-264
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    • 2023
  • This study was conducted to evaluate the applicability of the Comet assay, which is widely used for the identification of irradiated meats, to detect irradiated beef cut, ground beef, and Tteokgalbi during freezing storage. Gamma-irradiation significantly increased the DNA damage in frozen beef cut and ground beef samples. Among those, DNA nuclei of samples irradiated with absorbed doses of 1kGy or more showed typical comet-shaped damage, convincing that the samples were irradiated. Meanwhile, DNA nuclei in non-irradiated beef cut and ground beef samples were also damaged according to storage time. In particular, since the damage of DNA nuclei in the non-irradiated samples frozen for three months was similar to that of samples irradiated with a dose of 0.5 kGy, it was considered difficult to detect whether these samples were irradiated by Comet assay analysis. Likewise, gamma-irradiation of Tteokgalbi increased DNA damage. However, significant damage to DNA nuclei was observed even in the non-irradiated samples. Therefore, the application of the analysis method to determine whether the Tteokgalbi sample was irradiated was not appropriate. In conclusion, these results suggest that Comet assay could be limitedly applied only to fresh meat with a short storage period and minimal processing.

ChIP-seq Library Preparation and NGS Data Analysis Using the Galaxy Platform (ChIP-seq 라이브러리 제작 및 Galaxy 플랫폼을 이용한 NGS 데이터 분석)

  • Kang, Yujin;Kang, Jin;Kim, Yea Woon;Kim, AeRi
    • Journal of Life Science
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    • v.31 no.4
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    • pp.410-417
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    • 2021
  • Next-generation sequencing (NGS) is a high-throughput technique for sequencing large numbers of DNA fragments that are prepared from a genome. This sequencing technique has been used to elucidate whole genome sequences of living organisms and to analyze complementary DNA (cDNA) or chromatin immunoprecipitated DNA (ChIPed DNA) at the genome level. After NGS, the use of proper tools is important for processing and analyzing data with reasonable parameters. However, handling large-scale sequencing data and programing for data analysis can be difficult. The Galaxy platform, a public web service system, provides many different tools for NGS data analysis, and it allows researchers to analyze their data on a web browser with no deep knowledge about bioinformatics and/or programing. In this study, we explain the procedure for preparing chromatin immunoprecipitation-sequencing (ChIP-seq) libraries and steps for analyzing ChIP-seq data using the Galaxy platform. The data analysis steps include the NGS data upload to Galaxy, quality check of the NGS data, premapping processes, read mapping, the post-mapping process, peak-calling and visualization by window view, heatmaps, average profile, and correlation analysis. Analysis of our histone H3K4me1 ChIP-seq data in K562 cells shows that it correlates with public data. Thus, NGS data analysis using the Galaxy platform can provide an easy approach to bioinformatics.

Development of Robot System for Colony Picking (I) - Image processing algorithm for detecting colony - (콜로니 픽킹 로봇 시스템의 개발 (I) - 콜로니 검출 영상처리 알고리즘 -)

  • 이현동;김기대;나건영;임용표
    • Journal of Biosystems Engineering
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    • v.28 no.5
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    • pp.439-448
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    • 2003
  • An image processing algorithm was developed for a robot system which was used in gene study. The robot system achieved a job of colony picking. The colony included DNA of an organism. The robot picked up the colony in petri-dish, which included the cultivated colony in medium, by a picking pin, and moved the colony to wellplates. The vision system consisted of an image acquisition system which acquired the image information of colony, an illumination device which irradiated the object once when it got the image of it, a computer and so on. The image processing algorithm distinguished the colony and detected colony positions. Performance test of the developed algorithm showed that the distinguishing success rate of colony and detecting success rate of colony positions were over 96%.

Expression of Lymphocyte ADP-ribosyltransferase in Rat Mammary Adenocarcinoma Cells (임파구 ADP-ribosyltransferase의 rat mammary adenocarcinoma cell에서의 발현)

  • 김현주
    • Journal of Life Science
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    • v.8 no.1
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    • pp.102-108
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    • 1998
  • The nascent from of glycosylphosphatidylinositol (GPI)-anchored proteins possesses both amino and carboxy terminal hydrophobic signal sequences to direct processing in the endoplasmic reticulum (ER). Following cleavage of the amino-terminal signal peptide, the carboxy-terminal peptide is processed. Previously, mouse lymphocyte NDA: agrinine ADP-ribosyltransferase (Yac-1) was cloned and the deduced amino acid sequence of the Yac-1 transferase contained hydrophobic amino and carboxy termini, consistent with known signal sequences of GPI-anchored proteins. This tranferase was present on the surface of NMU (rat mammary adenocarcinoma) cells transfected with the wildtype cDNA and was released with phosphatidylinositol-specific phosphilpase C. Expression of the mutant protein, lacking the carboxy terminal hydrophobic sequence, resulted in the peoduction of soluble, secreted from of the transferase. This result shows that carboxy terminal sequence is important for GPI-attachment.

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Sequential Pattern Mining with Optimization Calling MapReduce Function on MapReduce Framework (맵리듀스 프레임웍 상에서 맵리듀스 함수 호출을 최적화하는 순차 패턴 마이닝 기법)

  • Kim, Jin-Hyun;Shim, Kyu-Seok
    • The KIPS Transactions:PartD
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    • v.18D no.2
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    • pp.81-88
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    • 2011
  • Sequential pattern mining that determines frequent patterns appearing in a given set of sequences is an important data mining problem with broad applications. For example, sequential pattern mining can find the web access patterns, customer's purchase patterns and DNA sequences related with specific disease. In this paper, we develop the sequential pattern mining algorithms using MapReduce framework. Our algorithms distribute input data to several machines and find frequent sequential patterns in parallel. With synthetic data sets, we did a comprehensive performance study with varying various parameters. Our experimental results show that linear speed up can be achieved through our algorithms with increasing the number of used machines.

Gene Analysis Related Energy Metabolism of Leaf Expressed Sequence Tags Database of Korean Ginseng (Panax ginseng C.A. Meyer) (고려인삼(Panax ginseng C.A, Meyer)의 잎 ESTs database에서 Energy 대사 관련 유전자 분석)

  • Lee Jong-Il;Yoon Jae-Ho;Song Won-Seob;Lee Bum-Soo;In Jun-Gyo;Kim Eun-Jeong;Yang Deok-Chun
    • Korean Journal of Plant Resources
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    • v.19 no.1
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    • pp.174-179
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    • 2006
  • A cDNA library was constructed from leaf samples of 4-year-old Panax ginseng cultured in a field. 3,000 EST from a size selected leaf cDNA library were analyzed. The 349 of 2,896 cDNA clones has related with energy metabolism genes. The 349 known genes were categorized into nine groups according to their functional classification, aerobic respiration(48.4%), accessory proteins of electron transport and membrane associated energy conservation(17.2%), glycolysis and gluconeogenesis(3.4%), electron transport and membrane associated energy conservation(2.9%), respiration(2.0%), glycolysis methylglyoxal bypass(1.7%), metabolism of energy reserves(0.6%) and alcohol fermentation(0.3%).