• Title/Summary/Keyword: Genome type

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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.

Prediction of an Essential Gene with Potential Drug Target Property in Streptococcus suis Using Comparative Genomics

  • Zaman, Aubhishek
    • Interdisciplinary Bio Central
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    • v.4 no.4
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    • pp.11.1-11.8
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    • 2012
  • Genes that are indispensable for survival are referred to as essential gene. Due to the momentous significance of these genes for cellular activity they can be selected potentially as drug targets. Here in this study, an essential gene for Streptococcus suis was predicted using coherent statistical analysis and powerful genome comparison computational method. At first the whole genome protein scatter plot was generated and subsequently, on the basis of statistical significance, a reference genome was chosen. The parameters set forth for selecting the reference genome was that the genome of the query (Streptococcus suis) and subject must fall in the same genus and yet they must vary to a good degree. Streptococcus pneumoniae was found to be suitable as the reference genome. A whole genome comparison was performed for the reference (Streptococcus pneumoniae) and the query genome (Streptococcus suis) and 14 conserved proteins from them were subjected to a screen for potential essential gene property. Among those 14 only one essential gene was found to be with impressive similarity score between reference and query. The essential gene encodes for a type of 'Clp protease'. Clp proteases play major roles in degrading misfolded proteins. Results found here should help formulating a drug against Strptococcus suis which is responsible for mild to severe clinical conditions in human. However, like many other computational studies, the study has to be validated furthermore through in vitro assays for concrete proof.

Impact of type 2 diabetes variants identified through genome-wide association studies in early-onset type 2 diabetes from South Indian population

  • Liju, Samuel;Chidambaram, Manickam;Mohan, Viswanathan;Radha, Venkatesan
    • Genomics & Informatics
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    • v.18 no.3
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    • pp.27.1-27.12
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    • 2020
  • The prevalence of early-onset type 2 diabetes (EOT2D) is increasing in Asian countries. Genome-wide association studies performed in European and various other populations have identified associations of numerous variants with type 2 diabetes in adults. However, the genetic component of EOT2D which is still unexplored could have similarities with late-onset type 2 diabetes. Here in the present study we aim to identify the association of variants with EOT2D in South Indian population. Twenty-five variants from 18 gene loci were genotyped in 1,188 EOT2D and 1,183 normal glucose tolerant subjects using the MassARRAY technology. We confirm the association of the HHEX variant rs1111875 with EOT2D in this South Indian population and also the association of CDKN2A/2B (rs7020996) and TCF7L2 (rs4506565) with EOT2D. Logistic regression analyses of the TCF7L2 variant rs4506565(A/T), showed that the heterozygous and homozygous carriers for allele 'T' have odds ratios of 1.47 (95% confidence interval [CI], 1.17 to 1.83; p = 0.001) and 1.65 (95% CI, 1.18 to 2.28; p = 0.006) respectively, relative to AA homozygote. For the HHEX variant rs1111875 (T/C), heterozygous and homozygous carriers for allele 'C' have odds ratios of 1.13 (95% CI, 0.91 to 1.42; p = 0.27) and 1.58 (95% CI, 1.17 to 2.12; p = 0.003) respectively, relative to the TT homozygote. For CDKN2A/2B variant rs7020996, the heterozygous and homozygous carriers of allele 'C' were protective with odds ratios of 0.65 (95% CI, 0.51 to 0.83; p = 0.0004) and 0.62 (95% CI, 0.27 to 1.39; p = 0.24) respectively, relative to TT homozygote. This is the first study to report on the association of HHEX variant rs1111875 with EOT2D in this population.

In Silico Identification of 6-Phosphogluconolactonase Genes that are Frequently Missing from Completely Sequenced Bacterial Genomes

  • Jeong, Hae-Young;F. Kim, Ji-Hyun;Park, Hong-Seog
    • Genomics & Informatics
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    • v.4 no.4
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    • pp.182-187
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    • 2006
  • 6-Phosphogluconolactonase (6PGL) is one of the key enzymes in the ubiquitous pathways of central carbon metabolism, but bacterial 6PGL had been long known as a missing enzyme even after complete bacterial genome sequence information became available. Although recent experimental characterization suggests that there are two types of 6PGLs (DevB and YbhE), their phylogenetic distribution is severely biased. Here we present that proteins in COG group previously described as 3-oarboxymuconate cyclase (COG2706) are actually the YbhE-type 6PGLs, which are widely distributed in Proteobacteria and Fimicutes. This case exemplifies how erroneous functional description of a member in the reference database commonly used in transitive genome annotation cause systematic problem in the prediction of genes even with universal cellular functions.

Whole Genome Sequence of Streptomyces sp. from Novel Marine Actinomycetes

  • Hyeon Kyeong Lee;Heung-Soon Park;Eung-Soo Kim;Si-Sun Choi
    • Microbiology and Biotechnology Letters
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    • v.51 no.3
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    • pp.325-327
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    • 2023
  • This paper presents the complete genome sequence of a novel marine actinomycete, Streptomyces sp. MMBL 11-1. The genome of Streptomyces sp. MMBL 11-1 was obtained through next-generation sequencing using the PacBio Sequel system and Illumina platform provided by Macrogen, Korea. The assembled genome consists of five contigs, with a total length of 8,496,900 bp and a G+C content of 71.6%. The genome harbors multiple biosynthetic gene clusters (BGCs) associated with producing microbial natural products (MNPs). The comprehensive genomic information of this type of strain will serve as a valuable resource for identifying other marine actinomycetes strains.

Measuring and Reducing Off-Target Activities of Programmable Nucleases Including CRISPR-Cas9

  • Koo, Taeyoung;Lee, Jungjoon;Kim, Jin-Soo
    • Molecules and Cells
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    • v.38 no.6
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    • pp.475-481
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    • 2015
  • Programmable nucleases, which include zinc-finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and RNA-guided engineered nucleases (RGENs) repurposed from the type II clustered, regularly interspaced short palindromic repeats (CRISPR)-CRISPR-associated protein 9 (Cas9) system are now widely used for genome editing in higher eukaryotic cells and whole organisms, revolutionising almost every discipline in biological research, medicine, and biotechnology. All of these nucleases, however, induce off-target mutations at sites homologous in sequence with on-target sites, limiting their utility in many applications including gene or cell therapy. In this review, we compare methods for detecting nuclease off-target mutations. We also review methods for profiling genome-wide off-target effects and discuss how to reduce or avoid off-target mutations.

Draft genome sequence of Bacteroides sp. KGMB 02408 isolated from a healthy Korean feces (건강한 한국인 분변으로부터 분리된 Bacteroides sp. KGMB 02408 균주의 유전체 염기서열 초안)

  • Yu, Seung Yeob;Kim, Ji-Sun;Oh, Byeong Seob;Ryu, Seoung Woo;Park, Seung-Hwan;Kang, Se Won;Park, Jam-Eon;Choi, Seung-Hyeon;Han, Kook-Il;Lee, Keun Chul;Eom, Mi Kyung;Suh, Min Kuk;Kim, Han Sol;Lee, Dong Ho;Yoon, Hyuk;Kim, Byung-Yong;Lee, Je Hee;Lee, Jung-Sook;Lee, Ju Huck
    • Korean Journal of Microbiology
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    • v.55 no.3
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    • pp.296-299
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    • 2019
  • The genus of Bacteroides has been isolated from vertebrate animal feces. Bacteroides sp. KGMB 02408 was isolated from fecal samples obtained from a healthy Korean. The wholegenome sequence of Bacteroides sp. KGMB 02408 was analyzed using the PacBio Sequel platform. The genome comprises a 5,771,427 bp chromosome with a G + C content of 39.50%, 5,005 total genes, 18 rRNA genes, and 74 tRNA genes. Furthermore, we found that strain KGMB 02408 had some genes for oxidoreductases and menaquinone biosynthesis in its genome based on the result of genome analysis.

Cloning of Thymidine Kinase Gene of Herpes simplex Virus Type-1 (Herpes simplex Virus Type-1 Thymidine Kinase 유전자의 크로닝)

  • Kang, Hyun;Park, Kap-Joo;Cha, Sung-Chul;Kim, Soo-Yung;Yang, Ki-Sang;Kim, Nam-Joo;Lee, Hyung-Hoan
    • The Journal of Korean Society of Virology
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    • v.26 no.1
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    • pp.121-129
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    • 1996
  • Multiplication of Herpes simplex virus type-1 was observed by electronmicroscopy, a gene library of the genome was constructed and thymidine kinase gene was cloned. Vero cells infected with the virus were lysed 48 h p.j. and multinucleated giant cells were observed approximately at 72 h p.i. The nucleocapsids were observed in nuclei and cytoplasm, and the assembled nucleocapsids were budded out through the vacuole and cytoplasmic membranes, and then virions were released from the cells. HSV-1 genome DNA was digested with BamHI and BglII enzymes and then the gene library of the genome fragments were constructed. The BamHI cleaved the genome DNA into twenty-seven fragments in the range of 1.1 - 14 kb, and BglII cleaved the genome DNA into sixteen fragments in the range of $4.5{\sim}20.1\;kb$. The pHLA-12 and pHLB-4 recombinant plasmids were contained TK gene by Southern blot analysis. The molecular sizes of the fragments which contained the TK gene were 3.74 in pHLA-12 and 6.41kb in pHLB-4 recombinant plasmid, respectively.

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Considerations on gene chip data analysis

  • Lee, Jae-K.
    • Proceedings of the Korean Society for Bioinformatics Conference
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    • 2001.08a
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    • pp.77-102
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    • 2001
  • Different high-throughput chip technologies are available for genome-wide gene expression studies. Quality control and prescreening analysis are important for rigorous analysis on each type of gene expression data. Statistical significance evaluation of differential expression patterns is needed. Major genome institutes develop database and analysis systems for information sharing of precious expression data.

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No excessive mutations in transcription activator-like effector nuclease-mediated α-1,3-galactosyltransferase knockout Yucatan miniature pigs

  • Choi, Kimyung;Shim, Joohyun;Ko, Nayoung;Park, Joonghoon
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.2
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    • pp.360-372
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    • 2020
  • Objective: Specific genomic sites can be recognized and permanently modified by genome editing. The discovery of endonucleases has advanced genome editing in pigs, attenuating xenograft rejection and cross-species disease transmission. However, off-target mutagenesis caused by these nucleases is a major barrier to putative clinical applications. Furthermore, off-target mutagenesis by genome editing has not yet been addressed in pigs. Methods: Here, we generated genetically inheritable α-1,3-galactosyltransferase (GGTA1) knockout Yucatan miniature pigs by combining transcription activator-like effector nuclease (TALEN) and nuclear transfer. For precise estimation of genomic mutations induced by TALEN in GGTA1 knockout pigs, we obtained the whole-genome sequence of the donor cells for use as an internal control genome. Results: In-depth whole-genome sequencing analysis demonstrated that TALEN-mediated GGTA1 knockout pigs had a comparable mutation rate to homologous recombination-treated pigs and wild-type strain controls. RNA sequencing analysis associated with genomic mutations revealed that TALEN-induced off-target mutations had no discernable effect on RNA transcript abundance. Conclusion: Therefore, TALEN appears to be a precise and safe tool for generating genomeedited pigs, and the TALEN-mediated GGTA1 knockout Yucatan miniature pigs produced in this study can serve as a safe and effective organ and tissue resource for clinical applications.