• Title/Summary/Keyword: Genome engineering

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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.

Performance Comparison of Two Gene Set Analysis Methods for Genome-wide Association Study Results: GSA-SNP vs i-GSEA4GWAS

  • Kwon, Ji-Sun;Kim, Ji-Hye;Nam, Doug-U;Kim, Sang-Soo
    • Genomics & Informatics
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    • v.10 no.2
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    • pp.123-127
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    • 2012
  • Gene set analysis (GSA) is useful in interpreting a genome-wide association study (GWAS) result in terms of biological mechanism. We compared the performance of two different GSA implementations that accept GWAS p-values of single nucleotide polymorphisms (SNPs) or gene-by-gene summaries thereof, GSA-SNP and i-GSEA4GWAS, under the same settings of inputs and parameters. GSA runs were made with two sets of p-values from a Korean type 2 diabetes mellitus GWAS study: 259,188 and 1,152,947 SNPs of the original and imputed genotype datasets, respectively. When Gene Ontology terms were used as gene sets, i-GSEA4GWAS produced 283 and 1,070 hits for the unimputed and imputed datasets, respectively. On the other hand, GSA-SNP reported 94 and 38 hits, respectively, for both datasets. Similar, but to a lesser degree, trends were observed with Kyoto Encyclopedia of Genes and Genomes (KEGG) gene sets as well. The huge number of hits by i-GSEA4GWAS for the imputed dataset was probably an artifact due to the scaling step in the algorithm. The decrease in hits by GSA-SNP for the imputed dataset may be due to the fact that it relies on Z-statistics, which is sensitive to variations in the background level of associations. Judicious evaluation of the GSA outcomes, perhaps based on multiple programs, is recommended.

Prediction Model for the Cellular Immortalization and Transformation Potentials of Cell Substrates

  • Lee, Min-Su;Matthews Clayton A.;Chae Min-Ju;Choi, Jung-Yun;Sohn Yeo-Won;Kim, Min-Jung;Lee, Su-Jae;Park, Woong-Yang
    • Genomics & Informatics
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    • v.4 no.4
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    • pp.161-166
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    • 2006
  • The establishment of DNA microarray technology has enabled high-throughput analysis and molecular profiling of various types of cancers. By using the gene expression data from microarray analysis we are able to investigate diagnostic applications at the molecular level. The most important step in the application of microarray technology to cancer diagnostics is the selection of specific markers from gene expression profiles. In order to select markers of Immortalization and transformation we used c-myc and $H-ras^{V12}$ oncogene-transfected NIH3T3 cells as our model system. We have identified 8751 differentially expressed genes in the immortalization/transformation model by multivariate permutation F-test (95% confidence, FDR<0.01). Using the support vector machine algorithm, we selected 13 discriminative genes which could be used to predict immortalization and transformation with perfect accuracy. We assayed $H-ras^{V12}$-transfected 'transformed' cells to validate our immortalization/transformation dassification system. The selected molecular markers generated valuable additional information for tumor diagnosis, prognosis and therapy development.

Genome Sequence of Bacillus cereus FORC_021, a Food-Borne Pathogen Isolated from a Knife at a Sashimi Restaurant

  • Chung, Han Young;Lee, Kyu-Ho;Ryu, Sangryeol;Yoon, Hyunjin;Lee, Ju-Hoon;Kim, Hyeun Bum;Kim, Heebal;Jeong, Hee Gon;Choi, Sang Ho;Kim, Bong-Soo
    • Journal of Microbiology and Biotechnology
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    • v.26 no.12
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    • pp.2030-2035
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    • 2016
  • Bacillus cereus causes food-borne illness through contaminated foods; therefore, its pathogenicity and genome sequences have been analyzed in several studies. We sequenced and analyzed B. cereus strain FORC_021 isolated from a sashimi restaurant. The genome sequence consists of 5,373,294 bp with 35.36% GC contents, 5,350 predicted CDSs, 42 rRNA genes, and 107 tRNA genes. Based on in silico DNA-DNA hybridization values, B. cereus ATCC $14579^T$ was closest to FORC_021 among the complete genome-sequenced strains. Three major enterotoxins were detected in FORC_021. Comparative genomic analysis of FORC_021 with ATCC $14579^T$ revealed that FORC_021 harbored an additional genomic region encoding virulence factors, such as putative ADP-ribosylating toxin, spore germination protein, internalin, and sortase. Furthermore, in vitro cytotoxicity testing showed that FORC_021 exhibited a high level of cytotoxicity toward INT-407 human epithelial cells. This genomic information of FORC_021 will help us to understand its pathogenesis and assist in managing food contamination.

Novel Discovery of LINE-1 in a Korean Individual by a Target Enrichment Method

  • Shin, Wonseok;Mun, Seyoung;Kim, Junse;Lee, Wooseok;Park, Dong-Guk;Choi, Seungkyu;Lee, Tae Yoon;Cha, Seunghee;Han, Kyudong
    • Molecules and Cells
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    • v.42 no.1
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    • pp.87-95
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    • 2019
  • Long interspersed element-1 (LINE-1 or L1) is an autonomous retrotransposon, which is capable of inserting into a new region of genome. Previous studies have reported that these elements lead to genomic variations and altered functions by affecting gene expression and genetic networks. Mounting evidence strongly indicates that genetic diseases or various cancers can occur as a result of retrotransposition events that involve L1s. Therefore, the development of methodologies to study the structural variations and interpersonal insertion polymorphisms by L1 element-associated changes in an individual genome is invaluable. In this study, we applied a systematic approach to identify human-specific L1s (i.e., L1Hs) through the bioinformatics analysis of high-throughput next-generation sequencing data. We identified 525 candidates that could be inferred to carry non-reference L1Hs in a Korean individual genome (KPGP9). Among them, we randomly selected 40 candidates and validated that approximately 92.5% of non-reference L1Hs were inserted into a KPGP9 genome. In addition, unlike conventional methods, our relatively simple and expedited approach was highly reproducible in confirming the L1 insertions. Taken together, our findings strongly support that the identification of non-reference L1Hs by our novel target enrichment method demonstrates its future application to genomic variation studies on the risk of cancer and genetic disorders.

Genome-wide association study for loin muscle area of commercial crossbred pigs

  • Menghao Luan;Donglin Ruan;Yibin Qiu;Yong Ye;Shenping Zhou;Jifei Yang;Ying Sun;Fucai Ma;Zhenfang Wu;Jie Yang;Ming Yang;Enqin Zheng;Gengyuan Cai;Sixiu Huang
    • Animal Bioscience
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    • v.36 no.6
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    • pp.861-868
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    • 2023
  • Objective: Loin muscle area (LMA) is an important target trait of pig breeding. This study aimed to identify single nucleotide polymorphisms (SNPs) and genes associated with LMA in the Duroc×(Landrace×Yorkshire) crossbred pigs (DLY). Methods: A genome-wide association study was performed using the Illumina 50K chip to map the genetic marker and genes associated with LMA in 511 DLY pigs (255 boars and 256 sows). Results: After quality control, we detected 35,426 SNPs, including six SNPs significantly associated with LMA in pigs, with MARC0094338 and ASGA0072817 being the two key SNPs responsible for 1.77% and 2.48% of the phenotypic variance of LMA, respectively. Based on previous research, we determined two candidate genes (growth hormone receptor [GHR] and 3-oxoacid Co A-transferase 1 [OXCT1]) that are associated with fat deposition and muscle growth and found further additional genes (MYOCD, ARHGAP44, ELAC2, MAP2K4, FBXO4, FBLL1, RARS1, SLIT3, and RANK3) that are presumed to have an effect on LMA. Conclusion: This study contributes to the identification of the mutation that underlies quantitative trait loci associated with LMA and to future pig breeding programs based on marker-assisted selection. Further studies are needed to elucidate the role of the identified candidate genes in the physiological processes involved in LMA regulation.

Clustering and Comparative Analyses of Complete Genomes for the Elucidation of Evolutionary Characteristics

  • Kim, Jin-Sik;Lee, Sang-Yup
    • Proceedings of the Korean Society for Bioinformatics Conference
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    • 2005.09a
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    • pp.78-82
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    • 2005
  • Three of the genus Pseudomonas (P. aeruginosa, P. putida, P. syringae) show highly different phenotypic characteristics among them. Two of the three members are pathogenic and the other is non-pathogenic. Comparative analyses of the complete genomes can elucidate the genomic similarities and differences among them. We analyzed the three genomes and the genes of them to reveal the degree of conservation of chromosomes and similarity of the genes. The 2-dimensional dot plot between the pathogenic P. aeruginosa and non-pathogenic P. putida shared higher portion of the nucleotide sequences than other two combinations. Comparison of the nucleotide compositions by calculating the genome-scale plot of G+C contents and GC skew showed the variation of location. Comparison of the metabolic capabilities using the functional classification of KEGG orthology revealed that the differences in the number of genes for the specific functional categories resulted in the phenotypic differences. Finally combination of the analyses using the protein homologs supported the evolutionary distance of the P. putida obtained from other genome-scale comparisons.

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Computational identification of significantly regulated metabolic reactions by integration of data on enzyme activity and gene expression

  • Nam, Ho-Jung;Ryu, Tae-Woo;Lee, Ki-Young;Kim, Sang-Woo;Lee, Do-Heon
    • BMB Reports
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    • v.41 no.8
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    • pp.609-614
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    • 2008
  • The concentrations and catalytic activities of enzymes control metabolic rates. Previous studies have focused on enzyme concentrations because there are no genome-wide techniques used for the measurement of enzyme activity. We propose a method for evaluating the significance of enzyme activity by integrating metabolic network topologies and genome-wide microarray gene expression profiles. We quantified the enzymatic activity of reactions and report the 388 significant reactions in five perturbation datasets. For the 388 enzymatic reactions, we identified 70 that were significantly regulated (P-value < 0.001). Thirty-one of these reactions were part of anaerobic metabolism, 23 were part of low-pH aerobic metabolism, 8 were part of high-pH anaerobic metabolism, 3 were part of low-pH aerobic reactions, and 5 were part of high-pH anaerobic metabolism.

Draft genome sequence of lytic bacteriophage CF1 infecting Citrobacter freundii isolates (Citrobacter freundii 분리주를 감염시키는 용균 박테리오파지 CF1의 유전체 염기서열 초안)

  • Kim, Youngju;Ko, Seyoung;Yeon, Young Eun;Lim, Jaewon;Han, Beom Ku;Kim, Hyunil;Ahn, Jeong Keun;Kim, Donghyuk
    • Korean Journal of Microbiology
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    • v.54 no.1
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    • pp.79-80
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    • 2018
  • Citrobacter freundii is a facultative anaerobic and a Gram-negative bacterium of Enterobacteriaceae family, and is an opportunistic pathogen. Bacteriophages infecting C. freundii can be an effective treatment for C. freundii infections. Here, the complete genomic sequence is announced for a lytic bacteriophage CF1 infecting C. freundii isolates.

Outlook on genome editing application to cattle

  • Gyeong-Min Gim;Goo Jang
    • Journal of Veterinary Science
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    • v.25 no.1
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    • pp.10.1-10.11
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    • 2024
  • In livestock industry, there is growing interest in methods to increase the production efficiency of livestock to address food shortages, given the increasing global population. With the advancements in gene engineering technology, it is a valuable tool and has been intensively utilized in research specifically focused on human disease. In historically, this technology has been used with livestock to create human disease models or to produce recombinant proteins from their byproducts. However, in recent years, utilizing gene editing technology, cattle with identified genes related to productivity can be edited, thereby enhancing productivity in response to climate change or specific disease instead of producing recombinant proteins. Furthermore, with the advancement in the efficiency of gene editing, it has become possible to edit multiple genes simultaneously. This cattle breed improvement has been achieved by discovering the genes through the comprehensive analysis of the entire genome of cattle. The cattle industry has been able to address gene bottlenecks that were previously impossible through conventional breeding systems. This review concludes that gene editing is necessary to expand the cattle industry, improving productivity in the future. Additionally, the enhancement of cattle through gene editing is expected to contribute to addressing environmental challenges associated with the cattle industry. Further research and development in gene editing, coupled with genomic analysis technologies, will significantly contribute to solving issues that conventional breeding systems have not been able to address.