• Title/Summary/Keyword: whole-genome DNA

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Survey of the Applications of NGS to Whole-Genome Sequencing and Expression Profiling

  • Lim, Jong-Sung;Choi, Beom-Soon;Lee, Jeong-Soo;Shin, Chan-Seok;Yang, Tae-Jin;Rhee, Jae-Sung;Lee, Jae-Seong;Choi, Ik-Young
    • Genomics & Informatics
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    • v.10 no.1
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    • pp.1-8
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    • 2012
  • Recently, the technologies of DNA sequence variation and gene expression profiling have been used widely as approaches in the expertise of genome biology and genetics. The application to genome study has been particularly developed with the introduction of the nextgeneration DNA sequencer (NGS) Roche/454 and Illumina/ Solexa systems, along with bioinformation analysis technologies of whole-genome $de$ $novo$ assembly, expression profiling, DNA variation discovery, and genotyping. Both massive whole-genome shotgun paired-end sequencing and mate paired-end sequencing data are important steps for constructing $de$ $novo$ assembly of novel genome sequencing data. It is necessary to have DNA sequence information from a multiplatform NGS with at least $2{\times}$ and $30{\times}$ depth sequence of genome coverage using Roche/454 and Illumina/Solexa, respectively, for effective an way of de novo assembly. Massive shortlength reading data from the Illumina/Solexa system is enough to discover DNA variation, resulting in reducing the cost of DNA sequencing. Whole-genome expression profile data are useful to approach genome system biology with quantification of expressed RNAs from a wholegenome transcriptome, depending on the tissue samples. The hybrid mRNA sequences from Rohce/454 and Illumina/Solexa are more powerful to find novel genes through $de$ $novo$ assembly in any whole-genome sequenced species. The $20{\times}$ and $50{\times}$ coverage of the estimated transcriptome sequences using Roche/454 and Illumina/Solexa, respectively, is effective to create novel expressed reference sequences. However, only an average $30{\times}$ coverage of a transcriptome with short read sequences of Illumina/Solexa is enough to check expression quantification, compared to the reference expressed sequence tag sequence.

DNA Fragment Assembly

  • 박근수
    • Proceedings of the Korean Society for Bioinformatics Conference
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    • 2002.06a
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    • pp.105-121
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    • 2002
  • 최근 인간 지놈(genome)의 DNA가 밝혀져서 많은 관심을 받았는데, 이를 수행하는 방법을 소개한다. Human Genome Project에서 채택한 BAC-to-BAC 방식과 Celera 회사에서 채택한 whole genome shotgun 방식을 설명한다. 또한 두 방식에서 공히 fragment assembly 프로그램을 사용하는데, 이 프로그램의 개요를 설명한다.

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Development of a Rapid Foodborne-pathogen-detection Method Involving Whole-genome Amplification (Whole genome amplification을 이용한 식중독 세균 신속 검출 기술 개발)

  • Seong, Ji-Yeong;Ko, Young-Jun;Myeong, Hyeon-Koon;Oh, Se-Wook
    • Korean Journal of Food Science and Technology
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    • v.48 no.2
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    • pp.128-132
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    • 2016
  • In this study, polyethylene glycol (PEG) was used to improve DNA amplification efficiency during whole genome amplification (WGA). Amplification efficiency was determined by adding PEG with different molecular weights to the WGA reaction. The greatest increase in amplification efficiency was obtained with PEG 4,000 used at 1.5% concentration. Foodborne pathogenic DNA was amplified by WGA and quantitatively analyzed by real-time polymerase chain reaction. DNA of Salmonella serotype Typhimurium, Listeria monocytogenes, and Vibrio parahaemolyticus was amplified 7,777.01, 9,981.22, and 1,239.03 fold, respectively, by WGA. On adding PEG in the WGA reaction (i.e., enhanced WGA [eWGA]), 18-40-fold more DNA amplification was achieved. Thus, these analyses showed that foodborne pathogens, which are usually present at very low concentration in foods, can be detected by real-time PCR and WGA.

Analysis of genome variants in dwarf soybean lines obtained in F6 derived from cross of normal parents (cultivated and wild soybean)

  • Roy, Neha Samir;Ban, Yong-Wook;Yoo, Hana;Ramekar, Rahul Vasudeo;Cheong, Eun Ju;Park, Nam-Il;Na, Jong Kuk;Park, Kyong-Cheul;Choi, Ik-Young
    • Genomics & Informatics
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    • v.19 no.2
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    • pp.19.1-19.9
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    • 2021
  • Plant height is an important component of plant architecture and significantly affects crop breeding practices and yield. We studied DNA variations derived from F5 recombinant inbred lines (RILs) with 96.8% homozygous genotypes. Here, we report DNA variations between the normal and dwarf members of four lines harvested from a single seed parent in an F6 RIL population derived from a cross between Glycine max var. Peking and Glycine soja IT182936. Whole genome sequencing was carried out, and the DNA variations in the whole genome were compared between the normal and dwarf samples. We found a large number of DNA variations in both the dwarf and semi-dwarf lines, with one single nucleotide polymorphism (SNP) per at least 3.68 kb in the dwarf lines and 1 SNP per 11.13 kb of the whole genome. This value is 2.18 times higher than the expected DNA variation in the F6 population. A total of 186 SNPs and 241 SNPs were discovered in the coding regions of the dwarf lines 1282 and 1303, respectively, and we discovered 33 homogeneous nonsynonymous SNPs that occurred at the same loci in each set of dwarf and normal soybean. Of them, five SNPs were in the same positions between lines 1282 and 1303. Our results provide important information for improving our understanding of the genetics of soybean plant height and crop breeding. These polymorphisms could be useful genetic resources for plant breeders, geneticists, and biologists for future molecular biology and breeding projects.

DNA Chip Technologies

  • Hwang, Seoung-Yong;Lim, Geun-Bae
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.5 no.3
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    • pp.159-163
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    • 2000
  • The genome sequencing project has generated and will contitute to generate enormous amounts of sequence data. Since the first complete genome sequence of bacterium Haemophilus in fluenzae was published in 1995, the complete genome sequences of 2 eukaryotic and about 22 prokaryotic organisms have detemined. Given this everincreasing amounts of sequence information, new strategies are necessary to efficiently pursue the phase of the geome project- the elucidation of gene expression patterns and gene product function on a whole genome scale. In order to assign functional information to the genome sequence, DNA chip technology was developed to efficienfly identify the differential expression pattern of indepondent biogical samples. DNA chip provides a new tool for genome expreesion analysis that may revolutionize revolutionize many aspects of human kife including mew surg discovery and human disease diagnostics.

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Utilization of whole genome treasure for the library construction of industrial enzymes

  • Kim, Won-Ho;Cho, Kyoung-Won;Jung, In-Su;Choi, Keum-Hwa;Hur, Byung-Ki;Kim, Geun-Joong
    • 한국생물공학회:학술대회논문집
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    • 2003.10a
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    • pp.815-820
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    • 2003
  • A huge database resulted from whole genome sequencing has provided a possibility of new information that is likely to extent the scope and thus changes the way of approach for the functional assigning of putative open reading frames annotated by whole genome sequence analyses. These are mainly realized by ease, one-step identification of putative genes using genomics or proteomics tools. A major challenge remained in biotechnology may translate these informations into better ways to screen or select a gene as a representative sequence. Further attempts to mine the related whole genes or partial DNA fragment from whole genome treasure, and then the incorporation of these sequences into a representative template, will result in the use of putative genes that can be translated into functional proteins or allowed the generation of new lineages as a valuable pool. Such screens enable rapid biochemical analysis and easy isolation of the target activity, thereby accelerating the screening of novel enzymes from the expanded library with related sequences. Information-based PCR amplification of whole genes and reconstitution of functional DNA fragments will provide a platform for expanding the functional spaces of potential enzymes, especially when used mixed- or metagenome as gene resources.

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Genome Organization of Temperate Phage 11143 from Emetic Bacillus cereus NCTC11143

  • Lee, Young-Duck;Park, Jong-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.22 no.5
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    • pp.649-653
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    • 2012
  • A temperate phage was isolated from emetic Bacillus cereus NCTC 11143 by mitomycin C and characterized by transmission electron microscopy and DNA and protein analyses. Whole genome sequencing of Bacillus phage 11143 was performed by GS-FLX. The phage has a dsDNA genome of 39,077 bp and a 35% G+C content. Bioinformatic analysis of the phage genome revealed 49 putative ORFs involved in replication, morphogenesis, DNA packaging, lysogeny, and host lysis. Bacillus phage 11143 could be classified as a member of the Siphoviridae family by morphology and genome structure. Genomic comparisons at the DNA and protein levels revealed homologous genetic modules with patterns and morphogenesis proteins similar to those of other Bacillus phages. Thus, Bacillus phages might have a mosaic genetic relationship.

Analysis of Nuclear Mitochondrial DNA Segments of Nine Plant Species: Size, Distribution, and Insertion Loci

  • Ko, Young-Joon;Kim, Sangsoo
    • Genomics & Informatics
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    • v.14 no.3
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    • pp.90-95
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    • 2016
  • Nuclear mitochondrial DNA segment (Numt) insertion describes a well-known phenomenon of mitochondrial DNA transfer into a eukaryotic nuclear genome. However, it has not been well understood, especially in plants. Numt insertion patterns vary from species to species in different kingdoms. In this study, the patterns were surveyed in nine plant species, and we found some tip-offs. First, when the mitochondrial genome size is relatively large, the portion of the longer Numt is also larger than the short one. Second, the whole genome duplication event increases the ratio of the shorter Numt portion in the size distribution. Third, Numt insertions are enriched in exon regions. This analysis may be helpful for understanding plant evolution.

Complete Genome Sequence of Bifidobacterium bifidum DS0908, Isolated from Human Fecal Sample

  • Haneol Yang;Yong-Sik Kim;Doo-Sang Park
    • Microbiology and Biotechnology Letters
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    • v.51 no.4
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    • pp.566-568
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    • 2023
  • In this report, we present the whole-genome sequence of Bifidobacterium bifidum DS0908 isolated from the human fecal sample. The genome composed of a single circular chromosome is 2,223,317 bp long and the DNA G+C content is 62.65%. No virulence genes were detected in the genomic sequences of B. bifidum DS0908.

Prospect of plant molecular cytogenetics in the 21st century

  • Mukai, Yasuhiko
    • Proceedings of the Korean Society of Life Science Conference
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    • 2003.10a
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    • pp.14-27
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    • 2003
  • The genomes of Arabidopsis and rice have been fully sequenced. Genomic sequencing provides global information about genome structure and organization. A comprehensive research account of our recent studies conducted on genome painting, comparative genomics and genome fusion is provided in order to project the prospects of plant cytogenetic research in post-genomics era. Genome analysis by GISH using genome painting is demonstrated as an excellent means suitable for visualization of a whole genome, since total genomic DNA representing the overall molecular composition of the genome is used as a probe. FISH on extended DNA fibers has been developed for high-resolution FISH and has contributed to determining the copy number and order of genes. We have also mapped a number of genes involving starch synthesis on wheat chromosomes by FISH and compared the position of these genes on linkage map of rice. Macro synteny between wheat and rice can be observed by comparing the location of these genes in spite of the fact that the size of DNA per chromosome differs by 20 fold in two. Moreover, to approach our goal towards making bread and udon noodles from rice flour in future by incorporating bread making and the noodle qualifies in rice, we have been successful in introducing large genomic DNA fragments containing agronomically important genes of wheat into a rice by successive introduction of large insert BAC clones, there by expanding genetic variability in rice. We call this method genome fusion.

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