• Title/Summary/Keyword: NGS

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Genome-wide analysis of sequence variations in eight inbred watermelon lines (수박계통간 염색체수준의 유전적변이 분석)

  • Kim, Youn-Sung;Ko, Chan-Sup;Yang, Hee-Beom;Kang, Sun-Chul
    • Journal of Plant Biotechnology
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    • v.43 no.2
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    • pp.164-173
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    • 2016
  • To investigate the genetic basis of phenotypic differences, sequence variations were analyzed in 8 inbred watermelon lines by re-sequencing. The number of sequence variations differed depending on the chromosome. Only 12.9% of SNPs were found within genes, whereas the rest were detected in promoter or intergenic regions. SNP density analysis showed that there was a highly variable region at the end of chromosome 6, which is similar to previously published findings. However, this region with high SNP density did not show much variation between the lines. In contrast, highly conserved regions with a size of 6.5-10 Mb were found in chromosomes 10 and 11. Pathway analysis suggested that the DIMBOA (a natural antibiotic)-glucoside degradation pathway was significantly different between the lines, indicating that the eight lines may have different levels of pathogen resistance. Among the carbohydrate-related genes, the alpha-galactosidase gene was the most variable among the lines. Information from this study will be helpful in understanding the watermelon breeding process at the molecular level.

Muscle differentiation induced up-regulation of calcium-related gene expression in quail myoblasts

  • Park, Jeong-Woong;Lee, Jeong Hyo;Kim, Seo Woo;Han, Ji Seon;Kang, Kyung Soo;Kim, Sung-Jo;Park, Tae Sub
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.9
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    • pp.1507-1515
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    • 2018
  • Objective: In the poultry industry, the most important economic traits are meat quality and carcass yield. Thus, many studies were conducted to investigate the regulatory pathways during muscle differentiation. To gain insight of muscle differentiation mechanism during growth period, we identified and validated calcium-related genes which were highly expressed during muscle differentiation through mRNA sequencing analysis. Methods: We conducted next-generation-sequencing (NGS) analysis of mRNA from undifferentiated QM7 cells and differentiated QM7 cells (day 1 to day 3 of differentiation periods). Subsequently, we obtained calcium related genes related to muscle differentiation process and examined the expression patterns by quantitative reverse-transcription polymerase chain reaction (qRT-PCR). Results: Through RNA sequencing analysis, we found that the transcription levels of six genes (troponin C1, slow skeletal and cardiac type [TNNC1], myosin light chain 1 [MYL1], MYL3, phospholamban [PLN], caveolin 3 [CAV3], and calsequestrin 2 [CASQ2]) particularly related to calcium regulation were gradually increased according to days of myotube differentiation. Subsequently, we validated the expression patterns of calcium-related genes in quail myoblasts. These results indicated that TNNC1, MYL1, MYL3, PLN, CAV3, CASQ2 responded to differentiation and growth performance in quail muscle. Conclusion: These results indicated that calcium regulation might play a critical role in muscle differentiation. Thus, these findings suggest that further studies would be warranted to investigate the role of calcium ion in muscle differentiation and could provide a useful biomarker for muscle differentiation and growth.

Targeted Resequencing of 30 Genes Improves the Detection of Deleterious Mutations in South Indian Women with Breast and/or Ovarian Cancers

  • Rajkumar, Thangarajan;Meenakumari, Balaiah;Mani, Samson;Sridevi, Veluswami;Sundersingh, Shirley
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.13
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    • pp.5211-5217
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    • 2015
  • Background: We earlier used PCR-dHPLC for mutation analysis of BRCA1 and BRCA2. In this article we report application of targeted resequencing of 30 genes involved in hereditary cancers. Materials and Methods: A total of 91 patient samples were analysed using a panel of 30 genes in the Illumina HiScan SQ system. CLCBio was used for mapping reads to the reference sequences as well as for quality-based variant detection. All the deleterious mutations were then reconfirmed using Sanger sequencing. Kaplan Meier analysis was conducted to assess the effect of deleterious mutations on disease free and overall survival. Results: Seventy four of the 91 samples had been run earlier using the PCR-dHPLC and no deleterious mutations had been detected while 17 samples were tested for the first time. A total of 24 deleterious mutations were detected, 11 in BRCA1, 4 in BRCA2, 5 in p53, one each in RAD50, RAD52, ATM and TP53BP1. Some 19 deleterious mutations were seen in patients who had been tested earlier with PCR-dHPLC [19/74] and 5/17 in the samples tested for the first time, Together with our earlier detected 21 deleterious mutations in BRCA1 and BRCA2, we now had 45 mutations in 44 patients. BRCA1c.68_69delAG;p.Glu23ValfsX16 mutation was the most common, seen in 10/44 patients. Kaplan Meier survival analysis did not show any difference in disease free and overall survival in the patients with and without deleterious mutations. Conclusions: The NGS platform is more sensitive and cost effective in detecting mutations in genes involved in hereditary breast and/or ovarian cancers.

Whole-transcriptome analyses of the Sapsaree, a Korean natural monument, before and after exercise-induced stress

  • Kim, Ji-Eun;Choe, Junkyung;Lee, Jeong Hee;Kim, Woong Bom;Cho, Whan;Ha, Ji Hong;Kwon, Ki Jin;Han, Kook Il;Jo, Sung-Hwan
    • Journal of Animal Science and Technology
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    • v.58 no.4
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    • pp.17.1-17.7
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    • 2016
  • Background: The Sapsaree (Canis familiaris) is a Korean native dog that is very friendly, protective, and loyal to its owner, and is registered as a natural monument in Korea (number: 368). To investigate large-scale gene expression profiles and identify the genes related to exercise-induced stress in the Sapsaree, we performed whole-transcriptome RNA sequencing and analyzed gene expression patterns before and after exercise performance. Results: We identified 525 differentially expressed genes in ten dogs before and after exercise. Gene Ontology classification and KEGG pathway analysis revealed that the genes were mainly involved in metabolic processes, such as programmed cell death, protein metabolic process, phosphatidylinositol signaling system, and cation binding in cytoplasm. The ten Sapsarees could be divided into two groups based on the gene expression patterns before and after exercise. The two groups were significantly different in terms of their basic body type ($p{\leq}0.05$). Seven representative genes with significantly different expression patterns before and after exercise between the two groups were chosen and characterized. Conclusions: Body type had a significant effect on the patterns of differential gene expression induced by exercise. Whole-transcriptome sequencing is a useful method for investigating the biological characteristics of the Sapsaree and the large-scale genomic differences of canines in general.

Comparison of transcriptome analysis between red flash peach cultivar and white flash peach cultivar using next generation sequencing (Next generation sequencing 방법을 이용한 적육계 복숭아와 백육계 복숭아의 전사체 분석)

  • Kim, Se Hee;Nam, Eun Young;Cho, Kang-Hee;Shin, Il Sheob;Kim, Hyun Ran;Hwang, Hae Seong
    • Journal of Plant Biotechnology
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    • v.39 no.4
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    • pp.273-280
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    • 2012
  • Differences of gene expression between red flash peach cultivar and white flash peach cultivar were investigated by Nest-generation sequencing (NGS). EST from the red flash peach cultivar and white flash peach cultivar were selected for nucleotide sequence determination and homology searches. The levels of transcripts coding for proteins involved in pathogenesis related proteins, temperature stress, ethylene signal pathway were significantly higher in white flash peach cultivar than in red flash peach cultivar. On the other hand, the up-regulation of proteins involved in anthocyanin and flavonol biosynthesis and protein degradation and sorbitol metabolism were observed in red flash peach cultivar. Chalcone synthase was preferentially expressed in the red flesh peach cultivar, agreeing with the accumulation of anthocyanin and expression of other previously identified genes for anthocyanin biosynthesis. Anthocyanin pathway related genes CHS, F3H, DFR, LDOX, UFGT differentially expressed between red flash peach cultivar and white flash peach cultivar. These results suggest that red flash peach cultivar and white flash peach cultivar have different anthocyanin biosynthesis regulatory mechanisms.

The strategy and current status of Brassica rapa genome project (배추 유전체 염기서열 해독 전략과 현황)

  • Mun, Jeong-Hwan;Kwon, Soo-Jin;Park, Beom-Seok
    • Journal of Plant Biotechnology
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    • v.37 no.2
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    • pp.153-165
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    • 2010
  • Brassica rapa is considered an ideal candidate to act as a reference species for Brassica genomic studies. Among the three basic Brassica species, B. rapa (AA genome) has the smallest genome (529 Mbp), compared to B. nigra (BB genome, 632 Mbp) and B. oleracea (CC genome, 696 Mbp). There is also a large collection of available cultivars of B. rapa, as well as a broad array of B. rapa genomic resources available. Under international consensus, various genomic studies on B. rapa have been conducted, including the construction of a physical map based on 22.5X genome coverage, end sequencing of 146,000 BACs, sequencing of >150,000 expressed sequence tags, and successful phase 2 shotgun sequencing of 589 euchromatic region-tiling BACs based on comparative positioning with the Arabidopsis genome. These sequenced BACs mapped onto the B. rapa genome provide beginning points for genome sequencing of each chromosome. Applying this strategy, all of the 10 chromosomes of B. rapa have been assigned to the sequencing centers in seven countries, Korea, UK, China, India, Canada, Australia, and Japan. The two longest chromosomes, A3 and A9, have been sequenced except for several gaps, by NAAS in Korea. Meanwhile a China group, including IVF and BGI, performed whole genome sequencing with Illumina system. These Sanger and NGS sequence data will be integrated to assemble a draft sequence of B. rapa. The imminent B. rapa genome sequence offers novel insights into the organization and evolution of the Brassica genome. In parallel, the transfer of knowledge from B. rapa to other Brassica crops would be expected.

Development of Molecular Markers and Application for Breeding in Chinese Cabbage (배추의 분자 마커 개발 및 육종적 활용)

  • Kim, Ho-Il;Hong, Chang Pyo;Im, Subin;Choi, Su Ryun;Lim, Yong Pyo
    • Horticultural Science & Technology
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    • v.32 no.6
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    • pp.745-752
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    • 2014
  • Chinese cabbage (Brassica rapa L. ssp. pekinensis) is an economically important vegetable crop as a source of the traditional food Kimchi in Korea. Although many varieties exhibiting desirable traits have been developed by the conventional selective breeding approach, breeding related to abiotic or biotic stresses, such as a particular pests or diseases, or tolerance to climatic conditions, is likely to be slow. This could be helped by an efficient method for selection from various, rapidly-evolved genetic resources on the basis of molecular markers. In particular, the Brassica genome sequencing project enables genome-wide discovery of genes or genetic variants associated with agricultural traits. We here discuss the recent progress in the field of Chinese cabbage breeding with regard to the application of molecular markers.

Detecting Positive Selection of Korean Native Goat Populations Using Next-Generation Sequencing

  • Lee, Wonseok;Ahn, Sojin;Taye, Mengistie;Sung, Samsun;Lee, Hyun-Jeong;Cho, Seoae;Kim, Heebal
    • Molecules and Cells
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    • v.39 no.12
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    • pp.862-868
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    • 2016
  • Goats (Capra hircus) are one of the oldest species of domesticated animals. Native Korean goats are a particularly interesting group, as they are indigenous to the area and were raised in the Korean peninsula almost 2,000 years ago. Although they have a small body size and produce low volumes of milk and meat, they are quite resistant to lumbar paralysis. Our study aimed to reveal the distinct genetic features and patterns of selection in native Korean goats by comparing the genomes of native Korean goat and crossbred goat populations. We sequenced the whole genome of 15 native Korean goats and 11 crossbred goats using next-generation sequencing (Illumina platform) to compare the genomes of the two populations. We found decreased nucleotide diversity in the native Korean goats compared to the crossbred goats. Genetic structural analysis demonstrated that the native Korean goat and cross-bred goat populations shared a common ancestry, but were clearly distinct. Finally, to reveal the native Korean goat's selective sweep region, selective sweep signals were identified in the native Korean goat genome using cross-population extended haplotype homozygosity (XP-EHH) and a cross-population composite likelihood ratio test (XP-CLR). As a result, we were able to identify candidate genes for recent selection, such as the CCR3 gene, which is related to lumbar paralysis resistance. Combined with future studies and recent goat genome information, this study will contribute to a thorough understanding of the native Korean goat genome.

AGB (Ancestral Genome Browser): A Web Interface for Browsing Reconstructed Ancestral Genomes (AGB (Ancestral Genome Browser): 조상유전체 데이터의 시각적 열람을 위한 웹 인터페이스)

  • Lee, Daehwan;Lee, Jongin;Hong, Woon-Young;Jang, Eunji;Kim, Jaebum
    • Journal of KIISE
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    • v.42 no.12
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    • pp.1584-1589
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    • 2015
  • With the advancement of next-generation sequencing (NGS) technologies, various genome browsers have been introduced. Because existing browsers focus on comparison of the genomic data of extant species, however, there is a need for a genome browser for ancestral genomes and their evolution. In this paper, we introduce a genome browser, AGB (Ancestral Genome Browser), that displays ancestral genome data reconstructed from existing species. With AGB, it is possible to trace genomic variations that occurred during evolution in a simple and intuitive way. We explain the capability of AGB in terms of visualizing ancestral genomic information and evolutionary genomic variations. AGB is now available at http://bioinfo.konkuk.ac.kr/genomebrowser/.

Comparative chloroplast genomics and phylogenetic analysis of the Viburnum dilatatum complex (Adoxaceae) in Korea

  • PARK, Jongsun;XI, Hong;OH, Sang-Hun
    • Korean Journal of Plant Taxonomy
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    • v.50 no.1
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    • pp.8-16
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    • 2020
  • Complete chloroplast genome sequences provide detailed information about any structural changes of the genome, instances of phylogenetic reconstruction, and molecular markers for fine-scale analyses. Recent developments of next-generation sequencing (NGS) tools have led to the rapid accumulation of genomic data, especially data pertaining to chloroplasts. Short reads deposited in public databases such as the Sequence Read Archive of the NCBI are open resources, and the corresponding chloroplast genomes are yet to be completed. The V. dilatatum complex in Korea consists of four morphologically similar species: V. dilatatum, V. erosum, V. japonicum, and V. wrightii. Previous molecular phylogenetic analyses based on several DNA regions did not resolve the relationship at the species level. In order to examine the level of variation of the chloroplast genome in the V. dilatatum complex, raw reads of V. dilatatum deposited in the NCBI database were used to reconstruct the whole chloroplast genome, with these results compared to the genomes of V. erosum, V. japonicum, and three other species in Viburnum. These comparative genomics results found no significant structural changes in Viburnum. The degree of interspecific variation among the species in the V. dilatatum complex is very low, suggesting that the species of the complex may have been differentiated recently. The species of the V. dilatatum complex share large unique deletions, providing evidence of close relationships among the species. A phylogenetic analysis of the entire genome of the Viburnum showed that V. dilatatum is a sister to one of two accessions of V. erosum, making V. erosum paraphyletic. Given that the overall degree of variation among the species in the V. dilatatum complex is low, the chloroplast genome may not provide a phylogenetic signal pertaining to relationships among the species.