• Title/Summary/Keyword: Whole-genome sequencing

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Genome re-sequencing to identify single nucleotide polymorphism markers for muscle color traits in broiler chickens

  • Kong, H.R.;Anthony, N.B.;Rowland, K.C.;Khatri, B.;Kong, B.C.
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.1
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    • pp.13-18
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    • 2018
  • Objective: Meat quality including muscle color in chickens is an important trait and continuous selective pressures for fast growth and high yield have negatively impacted this trait. This study was conducted to investigate genetic variations responsible for regulating muscle color. Methods: Whole genome re-sequencing analysis using Illumina HiSeq paired end read method was performed with pooled DNA samples isolated from two broiler chicken lines divergently selected for muscle color (high muscle color [HMC] and low muscle color [LMC]) along with their random bred control line (RAN). Sequencing read data was aligned to the chicken reference genome sequence for Red Jungle Fowl (Galgal4) using reference based genome alignment with NGen program of the Lasergene software package. The potential causal single nucleotide polymorphisms (SNPs) showing non-synonymous changes in coding DNA sequence regions were chosen in each line. Bioinformatic analyses to interpret functions of genes retaining SNPs were performed using the ingenuity pathways analysis (IPA). Results: Millions of SNPs were identified and totally 2,884 SNPs (1,307 for HMC and 1,577 for LMC) showing >75% SNP rates could induce non-synonymous mutations in amino acid sequences. Of those, SNPs showing over 10 read depths yielded 15 more reliable SNPs including 1 for HMC and 14 for LMC. The IPA analyses suggested that meat color in chickens appeared to be associated with chromosomal DNA stability, the functions of ubiquitylation (UBC) and quality and quantity of various subtypes of collagens. Conclusion: In this study, various potential genetic markers showing amino acid changes were identified in differential meat color lines, that can be used for further animal selection strategy.

Efficiency to Discovery Transgenic Loci in GM Rice Using Next Generation Sequencing Whole Genome Re-sequencing

  • Park, Doori;Kim, Dongin;Jang, Green;Lim, Jongsung;Shin, Yun-Ji;Kim, Jina;Seo, Mi-Seong;Park, Su-Hyun;Kim, Ju-Kon;Kwon, Tae-Ho;Choi, Ik-Young
    • Genomics & Informatics
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    • v.13 no.3
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    • pp.81-85
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    • 2015
  • Molecular characterization technology in genetically modified organisms, in addition to how transgenic biotechnologies are developed now require full transparency to assess the risk to living modified and non-modified organisms. Next generation sequencing (NGS) methodology is suggested as an effective means in genome characterization and detection of transgenic insertion locations. In the present study, we applied NGS to insert transgenic loci, specifically the epidermal growth factor (EGF) in genetically modified rice cells. A total of 29.3 Gb (${\sim}72{\times}coverage$) was sequenced with a $2{\times}150bp$ paired end method by Illumina HiSeq2500, which was consecutively mapped to the rice genome and T-vector sequence. The compatible pairs of reads were successfully mapped to 10 loci on the rice chromosome and vector sequences were validated to the insertion location by polymerase chain reaction (PCR) amplification. The EGF transgenic site was confirmed only on chromosome 4 by PCR. Results of this study demonstrated the success of NGS data to characterize the rice genome. Bioinformatics analyses must be developed in association with NGS data to identify highly accurate transgenic sites.

Metagenomic analysis of viral genes integrated in whole genome sequencing data of Thai patients with Brugada syndrome

  • Suwalak Chitcharoen;Chureerat Phokaew;John Mauleekoonphairoj;Apichai Khongphatthanayothin;Boosamas Sutjaporn;Pharawee Wandee;Yong Poovorawan;Koonlawee Nademanee;Sunchai Payungporn
    • Genomics & Informatics
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    • v.20 no.4
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    • pp.44.1-44.13
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    • 2022
  • Brugada syndrome (BS) is an autosomal dominant inheritance cardiac arrhythmia disorder associated with sudden death in young adults. Thailand has the highest prevalence of BS worldwide, and over 60% of patients with BS still have unclear disease etiology. Here, we performed a new viral metagenome analysis pipeline called VIRIN and validated it with whole genome sequencing (WGS) data of HeLa cell lines and hepatocellular carcinoma. Then the VIRIN pipeline was applied to identify viral integration positions from unmapped WGS data of Thai males, including 100 BS patients (case) and 100 controls. Even though the sample preparation had no viral enrichment step, we can identify several virus genes from our analysis pipeline. The predominance of human endogenous retrovirus K (HERV-K) viruses was found in both cases and controls by blastn and blastx analysis. This study is the first report on the full-length HERV-K assembled genomes in the Thai population. Furthermore, the HERV-K integration breakpoint positions were validated and compared between the case and control datasets. Interestingly, Brugada cases contained HERV-K integration breakpoints at promoters five times more often than controls. Overall, the highlight of this study is the BS-specific HERV-K breakpoint positions that were found at the gene coding region "NBPF11" (n = 9), "NBPF12" (n = 8) and long non-coding RNA (lncRNA) "PCAT14" (n = 4) region. The genes and the lncRNA have been reported to be associated with congenital heart and arterial diseases. These findings provide another aspect of the BS etiology associated with viral genome integrations within the human genome.

Analysis of whole genome sequencing and virulence factors of Vibrio vulnificus 1908-10 isolated from sea water at Gadeok island coast

  • Hee-kyung Oh;Nameun Kim;Do-Hyung Kim;Hye-Young Shin;Eun-Woo Lee;Sung-Hwan Eom;Young-Mog Kim
    • Fisheries and Aquatic Sciences
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    • v.26 no.9
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    • pp.558-568
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    • 2023
  • Vibrio vulnificus is an aquatic bacterium causing septicemia and wound infection in humans. To understand this pathogen at the genomic level, it was performed whole genome sequencing of a cefoxitin-resistant strain, V. vulnificus 1908-10 possessing virulence-related genes (vvhA, viuB, and vcgC) isolated from Gadeok island coastal seawater in South Korea. The genome of V. vulnificus 1908-10 consisted of two circular contigs and no plasmid. The total genome size was estimated to be 5,018,425 bp with a guanine-cytosine (GC) content of 46.9%. We found 119 tRNA and 34 rRNA genes respectively in the genome, along with 4,352 predicted protein sequences. Virulence factor (VF) analysis further revealed that V. vulnificus 1908-10 possess various virulence genes in classes of adherence, antiphagocytosis, chemotaxis and motility, iron uptake, quorum sensing, secretion system, and toxin. In the comparison of the presence/absence of virulence genes, V. vulnificus 1908-10 had fur, hlyU, luxS, ompU, pilA, pilF, rtxA, rtxC, and vvhA. Of the 30 V. vulnificus comparative strains, 80% of the C-genotype strains have all of these genes, whereas 40% of the E-genotype strains have all of them. In particular, pilA were identified in 80% of the C-type strains and 40% of the E-type strains, showing more difference than other genes. Therefore, V. vulnificus 1908-10 had similar VF characteristics to those of type C strains. Multifunctional-autoprocessing repeats-in-toxin (MARTX) toxin of V. vulnificus 1908-10 contained 8 A-type repeats (GXXGXXXXXG), 25 B.1-type repeats (TXVGXGXX), 18 B2-type repeats (GGXGXDXXX), and 7 C-type repeats (GGXGXDXXX). The National Center for Biotechnology Information (NCBI) Basic Local Alignment Search Tool (BLAST) showed that the RtxA protein of V. vulnificus 1908-10 had the effector domain in the order of cross-liking domain (ACD)-C58_PaToxP-like domain- α/β hydrolase-C58_PaToxP-like domain.

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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Draft Genome Sequence of Weissella koreensis Strain HJ, a Probiotic Bacterium Isolated from Kimchi

  • Seung-Min Yang;Eiseul Kim;So-Yun Lee;Soyeong Mun;Hae Choon Chang;Hae-Yeong Kim
    • Microbiology and Biotechnology Letters
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    • v.51 no.1
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    • pp.128-131
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    • 2023
  • Here we report the draft genome sequence of Weissella koreensis strain HJ and genomic analysis of its key features. The genome consists of 1,427,571 bp with a GC content of 35.5%, and comprises 1,376 coding genes. In silico analysis revealed the absence of pathogenic factors within the genome. The genome harbors several genes that play an important role in the survival of the gastrointestinal tract. In addition, a type III polyketide synthase cluster was identified. Pangenome analysis identified 68 unique genes in W. koreensis strain HJ. The genome information of this strain provides the basis for understanding its probiotic properties.

High-throughput identification of chrysanthemum gene function and expression: An overview and an effective proposition

  • Nguyen, Toan Khac;Lim, Jin Hee
    • Journal of Plant Biotechnology
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    • v.48 no.3
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    • pp.139-147
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    • 2021
  • Since whole-genome duplication (WGD) of diploid Chrysanthemum nankingense and de novo assembly whole-genome of C. seticuspe have been obtained, they have afforded to perceive the diversity evolution and gene discovery in the improved investigation of chrysanthemum breeding. The robust tools of high-throughput identification and analysis of gene function and expression produce their vast importance in chrysanthemum genomics. However, the gigantic genome size and heterozygosity are also mentioned as the major obstacles preventing the chrysanthemum breeding practices and functional genomics analysis. Nonetheless, some of technological contemporaries provide scientific efficient and promising solutions to diminish the drawbacks and investigate the high proficient methods for generous phenotyping data obtaining and system progress in future perspectives. This review provides valuable strategies for a broad overview about the high-throughput identification, and molecular analysis of gene function and expression in chrysanthemum. We also contribute the efficient proposition about specific protocols for considering chrysanthemum genes. In further perspective, the proper high-throughput identification will continue to advance rapidly and advertise the next generation in chrysanthemum breeding.

Whole Genome Analysis of Human Papillomavirus Type 16 Multiple Infection in Cervical Cancer Patients

  • Chansaenroj, Jira;Theamboonlers, Apiradee;Junyangdikul, Pairoj;Swangvaree, Sukumarn;Karalak, Anant;Poovorawan, Yong
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.2
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    • pp.599-606
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    • 2012
  • The characterization of the whole genome of human papillomavirus type 16 (HPV16) from cervical cancer specimens with multiple infections in comparison with single infection samples as the oncogenic potential of the virus may differ. Cervical carcinoma specimens positive for HPV16 by PCR and INNO-LiPA were randomly selected for whole genome characterization. Two HPV16 single infection and six HPV16 multiple infection specimens were subjected to whole genome analysis by using conserved primers and subsequent sequencing. All HPV16 whole genomes from single infection samples clustered in the European (E) lineage while all multiple infection specimens belonged to the non-European lineage. The variations in nucleotide sequences in E6, E7, E2, L1 and Long control region (LCR) were evaluated. In the E6 region, amino acid changes at L83V were related to increased cancer progression. An amino acid variation N29S within the E7 oncoprotein significantly associated with severity of lesion was also discovered. In all three domains of the E2 gene non synonymous mutations were found. The L1 region showed various mutations which may be related to conformation changes of viral epitopes. Some transcription factor binding sites in the LCR region correlated to virulence were shown on GRE/1, TEF-1, YY14 and Oct-1. HPV16 European variant prone to single infection may harbor a major variation at L83V which significantly increases the risk for developing cervical carcinoma. HPV16 non-European variants prone to multiple infections may require many polymorphisms to enhance the risk of cervical cancer development.

Whole Genome Resequencing of Heugu (Korean Black Cattle) for the Genome-Wide SNP Discovery

  • Choi, Jung-Woo;Chung, Won-Hyong;Lee, Kyung-Tai;Choi, Jae-Won;Jung, Kyoung-Sub;Cho, Yongmin;Kim, Namshin;Kim, Tae-Hun
    • Food Science of Animal Resources
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    • v.33 no.6
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    • pp.715-722
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    • 2013
  • Heugu (Korea Black Cattle) is one of the indigenous cattle breeds in Korea; however there has been severe lack of genomic studies on the breed. In this study, we report the first whole genome resequencing of Heugu at higher sequence coverage using Illumina HiSeq 2000 platform. More than 153.6 Giga base pairs sequence was obtained, of which 97% of the reads were mapped to the bovine reference sequence assembly (UMD 3.1). The number of non-redundantly mapped sequence reads corresponds to approximately 28.9-fold coverage across the genome. From these data, we identified a total of over six million single nucleotide polymorphisms (SNPs), of which 29.4% were found to be novel using the single nucleotide polymorphism database build 137. Extensive annotation was performed on all the detected SNPs, showing that most of SNPs were located in intergenic regions (70.7%), which is well corresponded with previous studies. Of the total SNPs, we identified substantial numbers of non-synonymous SNPs (13,979) in 5,999 genes, which could potentially affect meat quality traits in cattle. These results provide genome-wide SNPs that can serve as useful genetic tools and as candidates in searches for phenotype-altering DNA difference implicated with meat quality traits in cattle. The importance of this study can be further pronounced with the first whole genome sequencing of the valuable local genetic resource to be used in further genomic comparison studies with diverse cattle breeds.

Unraveling Biohydrogen Production and Sugar Utilization Systems in the Electricigen Shewanella marisflavi BBL25

  • Sang Hyun Kim;Hyun Joong Kim;Su Hyun Kim;Hee Ju Jung;Byungchan Kim;Do-Hyun Cho;Jong-Min Jeon;Jeong-Jun Yoon;Sang-Hyoun Kim;Jeong-Hoon Park;Shashi Kant Bhatia;Yung-Hun Yang
    • Journal of Microbiology and Biotechnology
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    • v.33 no.5
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    • pp.687-697
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    • 2023
  • Identification of novel, electricity-producing bacteria has garnered remarkable interest because of the various applications of electricigens in microbial fuel cell and bioelectrochemical systems. Shewanella marisflavi BBL25, an electricity-generating microorganism, uses various carbon sources and shows broader sugar utilization than the better-known S. oneidensis MR-1. To determine the sugar-utilizing genes and electricity production and transfer system in S. marisflavi BBL25, we performed an in-depth analysis using whole-genome sequencing. We identified various genes associated with carbon source utilization and the electron transfer system, similar to those of S. oneidensis MR-1. In addition, we identified genes related to hydrogen production systems in S. marisflavi BBL25, which were different from those in S. oneidensis MR-1. When we cultured S. marisflavi BBL25 under anaerobic conditions, the strain produced 427.58 ± 5.85 µl of biohydrogen from pyruvate and 877.43 ± 28.53 µl from xylose. As S. oneidensis MR-1 could not utilize glucose well, we introduced the glk gene from S. marisflavi BBL25 into S. oneidensis MR-1, resulting in a 117.35% increase in growth and a 17.64% increase in glucose consumption. The results of S. marisflavi BBL25 genome sequencing aided in the understanding of sugar utilization, electron transfer systems, and hydrogen production systems in other Shewanella species.