• 제목/요약/키워드: Genome analysis

검색결과 2,394건 처리시간 0.027초

Molecular Marker Development for the Rapid Differentiation of Black Rot Causing Xanthomonas campestris pv. campestris Race 7

  • Yeo-Hyeon Kim;Sopheap Mao;Nihar Sahu;Uzzal Somaddar;Hoy-Taek Kim;Masao Watanabe;Jong-In Park
    • The Plant Pathology Journal
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    • 제39권5호
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    • pp.494-503
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    • 2023
  • Xanthomonas campestris pv. campestris (Xcc) is a plant pathogen of Brassica crops that causes black rot disease throughout the world. At present, 11 physiological races of Xcc (races 1-11) have been reported. The conventional method of using differential cultivars for Xcc race detection is not accurate and it is laborious and time-consuming. Therefore, the development of specific molecular markers has been used as a substitute tool because it offers an accurate and reliable result, particularly a quick diagnosis of Xcc races. Previously, our laboratory has successfully developed race-specific molecular markers for Xcc races 1-6. In this study, specific molecular markers to identify Xcc race 7 have been developed. In the course of study, whole genome sequences of several Xcc races, X. campestris pv. incanae, X. campestris pv. raphani, and X. campestris pv. vesicatoria were aligned to identify variable regions like sequence-characterized amplified regions and insertions and deletions specific to race 7. Primer pairs were designed targeting these regions and validated against 22 samples. The polymerase chain reaction analysis revealed that three primer pairs specifically amplified the DNA fragment corresponding to race 7. The obtained finding clearly demonstrates the efficiency of the newly developed markers in accurately detecting Xcc race 7 among the other races. These results indicated that the newly developed marker can successfully and rapidly detect Xcc race 7 from other races. This study represents the first report on the successful development of specific molecular markers for Xcc race 7.

Molecular characterization of H3N2 influenza A virus isolated from a pig by next generation sequencing in Korea

  • Oh, Yeonsu;Moon, Sung-Hyun;Ko, Young-Seung;Na, Eun-Jee;Tark, Dong-Seob;Oem, Jae-Ku;Kim, Won-Il;Rim, Chaekwang;Cho, Ho-Seong
    • 한국동물위생학회지
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    • 제45권1호
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    • pp.31-38
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    • 2022
  • Swine influenza (SI) is an important respiratory disease in pigs and epidemic worldwide, which is caused by influenza A virus (IAV) belonging to the family of Orthomyxoviridae. As seen again in the 2009 swine-origin influenza A H1N1 pandemic, pigs are known to be susceptible to swine, avian, and human IAVs, and can serve as a 'mixing vessel' for the generation of novel IAV variants. To this end, the emergence of swine influenza viruses must be kept under close surveillance. Herein, we report the isolation and phylogenetic study of a swine IAV, A/swine/Korea/21810/2021 (sw21810, H3N2 subtype). BLASTN sequence analysis of 8 gene segments of the isolated virus revealed a high degree of nucleotide similarity (94.76 to 100%) to porcine strains circulating in Korea and the United States. Out of 8 genome segments, the HA gene was closely related to that of isolates from cluster I. Additionally, the NA gene of the isolate belonged to a Korean Swine H1N1 origin, and the PB2, PB1, NP and NS genes of the isolate were grouped into that of the Triple reassortant swine H3N2 origin virus. The PA and M genes of the isolate belonged to 2009 Pandemic H1N1 lineage. Human infection with mutants was most common through contact with infected pigs. Our results suggest the need for periodic close monitoring of this novel swine H3N2 influenza virus from a public health perspective.

Identification and Expression of Retroviral Envelope Polyprotein in the Dogfish Squalus mitsukurii

  • Kim, Soo Cheol;Sumi, Kanij Rukshana;Choe, Myeong Rak;Kho, Kang Hee
    • 한국해양생명과학회지
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    • 제1권2호
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    • pp.88-94
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    • 2016
  • Determining the infection history of living organisms is essential for understanding the evolution of infection agents with their host, particularly for key aspects such as immunity. Viruses, which can spread between individuals and often cause disease, have been widely examined. The increasing availability of fish genome sequences has provided specific insights into the diversity and host distribution of retroviruses in fish. The shortspine spurdog (Squalus mitsukurii) is an important elasmobranch species; this medium-sized dogfish typically lives at depths of 100~500 m. However, the retroviral envelope polyprotein in dogfish has not been examined. Thus, the aim of the present study was to identify and analyze the retroviral envelope polyprotein in various tissues of dogfish. The 1334-base pair full-length novel cDNA of dogfish envelope polyprotein (dEnv) was obtained by 3' and 5'-rapid amplification of cDNA end analysis from S. mitsukurii. The open reading frame showed a complete coding sequence of 815 base pairs with a deduced peptide sequence of 183 amino acids that exhibited 34~50% identity with other fish and bird species. It was also expressed according to reverse transcription and real-time polymerase chain reaction in the kidney, liver, intestine, and lung, but not in the gill. This distribution can be assessed by identifying and analyzing endogenous retroviruses in fish, which consists of three main genes: gag, pol and env. Dogfish envelope polyprotein sequence is likely important in evolution and induces rearrangements, altering the regulatory and coding sequences. This is the first report of the identification and molecular characterization of retroviral envelope polyprotein in various tissues of S. mitsukurii.

Bioinformatic analyses reveal the prognostic significance and potential role of ankyrin 3 (ANK3) in kidney renal clear cell carcinoma

  • Keerakarn Somsuan;Siripat Aluksanasuwan
    • Genomics & Informatics
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    • 제21권2호
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    • pp.22.1-22.15
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    • 2023
  • Kidney renal clear cell carcinoma (KIRC) is one of the most aggressive cancer type of the urinary system. Metastatic KIRC patients have poor prognosis and limited therapeutic options. Ankyrin 3 (ANK3) is a scaffold protein that plays important roles in maintaining physiological function of the kidney and its alteration is implicated in many cancers. In this study, we investigated differential expression of ANK3 in KIRC using GEPIA2, UALCAN, and HPA databases. Survival analysis was performed by GEPIA2, Kaplan-Meier plotter, and OS-kirc databases. Genetic alterations of ANK3 in KIRC were assessed using cBioPortal database. Interaction network and functional enrichment analyses of ANK3-correlated genes in KIRC were performed using GeneMANIA and Shiny GO, respectively. Finally, the TIMER2.0 database was used to assess correlation between ANK3 expression and immune infiltration in KIRC. We found that ANK3 expression was significantly decreased in KIRC compared to normal tissues. The KIRC patients with low ANK3 expression had poorer survival outcomes than those with high ANK3 expression. ANK3 mutations were found in 2.4% of KIRC patients and were frequently co-mutated with several genes with a prognostic significance. ANK3-correlated genes were significantly enriched in various biological processes, mainly involved in peroxisome proliferator-activated receptor (PPAR) signaling pathway, in which positive correlations of ANK3 with PPARA and PPARG expressions were confirmed. Expression of ANK3 in KIRC was significantly correlated with infiltration level of B cell, CD8+ T cell, macrophage, and neutrophil. These findings suggested that ANK3 could serve as a prognostic biomarker and promising therapeutic target for KIRC.

Effective microbial molecular diagnosis of periodontitis-related pathogen Porphyromonas gingivalis from salivary samples using rgpA gene

  • Jinuk Jeong;Yunseok Oh;Junhyeon Jeon;Dong-Heon Baek;Dong Hee Kim;Kornsorn Srikulnath;Kyudong Han
    • Genomics & Informatics
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    • 제21권1호
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    • pp.13.1-13.8
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    • 2023
  • Importance of accurate molecular diagnosis and quantification of particular disease-related pathogenic microorganisms is highlighted as an introductory step to prevent and care for diseases. In this study, we designed a primer/probe set for quantitative real-time polymerase chain reaction (qRT-PCR) targeting rgpA gene, known as the specific virulence factor of periodontitis-related pathogenic bacteria 'Porphyromonas gingivalis', and evaluated its diagnostic efficiency by detecting and quantifying relative bacterial load of P. gingivalis within saliva samples collected from clinical subjects. As a result of qRT-PCR, we confirmed that relative bacterial load of P. gingivalis was detected and quantified within all samples of positive control and periodontitis groups. On the contrary, negative results were confirmed in both negative control and healthy groups. Additionally, as a result of comparison with next-generation sequencing (NGS)-based 16S metagenome profiling data, we confirmed relative bacterial load of P. gingivalis, which was not identified on bacterial classification table created through 16S microbiome analysis, in qRT-PCR results. It showed that an approach to quantifying specific microorganisms by applying qRT-PCR method could solve microbial misclassification issues at species level of an NGS-based 16S microbiome study. In this respect, we suggest that P. gingivalis-specific primer/probe set introduced in present study has efficient applicability in various oral healthcare industries, including periodontitis-related microbial molecular diagnosis field.

Insight from sirtuins interactome: topological prominence and multifaceted roles of SIRT1 in modulating immunity, aging, and cancer

  • Nur Diyana Zulkifli;Nurulisa Zulkifle
    • Genomics & Informatics
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    • 제21권2호
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    • pp.23.1-23.9
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    • 2023
  • The mammalian sirtuin family, consisting of SIRT1-SIRT7, plays a vital role in various biological processes, including cancer, diabetes, neurodegeneration, cardiovascular disease, cellular metabolism, and cellular homeostasis maintenance. Due to their involvement in these biological processes, modulating sirtuin activity seems promising to impact immuneand aging-related diseases, as well as cancer pathways. However, more understanding is required regarding the safety and efficacy of sirtuin-targeted therapies due to the complex regulatory mechanisms that govern their activity, particularly in the context of multiple targets. In this study, the interaction landscape of the sirtuin family was analyzed using a systems biology approach. A sirtuin protein-protein interaction network was built using the Cytoscape platform and analyzed using the NetworkAnalyzer and stringApp plugins. The result revealed the sirtuin family's association with numerous proteins that play diverse roles, suggesting a complex interplay between sirtuins and other proteins. Based on network topological and functional analysis, SIRT1 was identified as the most prominent among sirtuin family members, demonstrating that 25 of its protein partners are involved in cancer, 22 in innate immune response, and 29 in aging, with some being linked to a combination of two or more pathways. This study lays the foundation for the development of novel therapies that can target sirtuins with precision and efficacy. By illustrating the various interactions among the proteins in the sirtuin family, we have revealed the multifaceted roles of SIRT1 and provided a framework for their possible roles to be precisely understood, manipulated, and translated into therapeutics in the future.

A phylogenetic analysis of the Korean endemic species Paraphlomis koreana (Lamiaceae) inferred from nuclear and plastid DNA sequences

  • Eun-Kyeong HAN;Jung-Hyun KIM;Jin-Seok KIM;Chang Woo HYUN;Dong Chan SON;Gyu Young CHUNG;Amarsanaa GANTSETSEG;Jung-Hyun LEE;In-Su CHOI
    • 식물분류학회지
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    • 제53권2호
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    • pp.157-165
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    • 2023
  • Paraphlomis koreana (Lamiaceae) was newly named and added to Korean flora in 2014. Paraphlomis belongs to the tribe Paraphlomideae, along with Ajugoides and Matsumurella. However, a recent study has suggested that P. koreana is morphologically similar to Matsumurella chinensis, making them difficult to distinguish from each other. Therefore, we aimed to examine the phylogenetic placement of P. koreana within the tribe and compare its genetic relationship with M. chinensis. We sequenced an additional complete plastid genome for an individual of P. koreana and generated sequences of nuclear ribosomal (nr) DNA regions of internal and external transcribed spacers (ITS and ETS) for two individuals of P. koreana. Maximum likelihood analyses based on two nrDNA regions (ITS and ETS) and four plastid DNA markers (rpl16 intron, rpl32-trnL, rps16 intron, and trnL-F) covering 13 Paraphlomis species and M. chinensis were conducted. Phylogenetic analyses concordantly supported that P. koreana forms a monophyletic group with M. chinensis. Moreover, our study revealed that P. koreana includes nrDNA sequences of M. chinensis as minor intra-individual variants, suggesting that the genetic divergence between the two taxa is incomplete and may represent intraspecific variation rather than distinct species. In conclusion, our findings suggest that the independent species status of P. koreana within Paraphlomis should be reconsidered.

DNA Sequence Variation of Candidate Gene for Salt Tolerance in Soybean Mutant

  • Chang Yeok Moon;Byeong Hee Kang;Woon Ji Kim;Sreeparna Chowdhury;Sehee Kang;Seo Young Shin;Wonho Lee;Hyeon-Seok Lee;Bo-Keun Ha
    • 한국작물학회:학술대회논문집
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    • 한국작물학회 2022년도 추계학술대회
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    • pp.259-259
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    • 2022
  • Soil salinity is a major factor that reduces crop yields. The amount of soil affected by salinity is about 83 million hectares (FAO 2000), which is increasing due to the effects of climate change. In soybean [Glycine max (L.) Merr.], nutritional properties such as protein, starch, and sucrose content together with biomass and yield tends to reduce due to excessive salt. As a result of QTL mapping using the 169 F2:3 population from the KA-1285 (salt-tolerant) × Daepung (salt-sensitive) in a previous study, two major QTLs (Gm03_39796778 and Gm03_40600088) related to salt tolerance were found on chromosome 3. In this study, the CDS region of the Gmsalt3 gene was analyzed using the ABI 3730x1 DNA Analyzer (Macrogen, Korea). The sequence of Gmsalt3 gene in KA-1285 was compared with Williams 82.a4.vl and PI483463 (Glycine soja). Two transversions were found at exon6 in KA-1285 and PI483463. Currently, whole genome sequencing and variation analysis using the Illumine Novaseq 6000 machine (Illumina, USA) are in progress. The results of this study can provide useful molecular markers for the selection of salt-tolerant soybeans and can be used as basic data for future salt-tolerant gene research.

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Multi-epitope vaccine against drug-resistant strains of Mycobacterium tuberculosis: a proteome-wide subtraction and immunoinformatics approach

  • Md Tahsin Khan;Araf Mahmud;Md. Muzahidul Islam;Mst. Sayedatun Nessa Sumaia;Zeaur Rahim;Kamrul Islam;Asif Iqbal
    • Genomics & Informatics
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    • 제21권3호
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    • pp.42.1-42.23
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    • 2023
  • Mycobacterium tuberculosis (Mtb) is the causative agent of tuberculosis, one of the most deadly infections in humans. The emergence of multidrug-resistant and extensively drug-resistant Mtb strains presents a global challenge. Mtb has shown resistance to many frontline antibiotics, including rifampicin, kanamycin, isoniazid, and capreomycin. The only licensed vaccine, Bacille Calmette-Guerin, does not efficiently protect against adult pulmonary tuberculosis. Therefore, it is urgently necessary to develop new vaccines to prevent infections caused by these strains. We used a subtractive proteomics approach on 23 virulent Mtb strains and identified a conserved membrane protein (MmpL4, NP_214964.1) as both a potential drug target and vaccine candidate. MmpL4 is a non-homologous essential protein in the host and is involved in the pathogen-specific pathway. Furthermore, MmpL4 shows no homology with anti-targets and has limited homology to human gut microflora, potentially reducing the likelihood of adverse effects and cross-reactivity if therapeutics specific to this protein are developed. Subsequently, we constructed a highly soluble, safe, antigenic, and stable multi-subunit vaccine from the MmpL4 protein using immunoinformatics. Molecular dynamics simulations revealed the stability of the vaccine-bound Tolllike receptor-4 complex on a nanosecond scale, and immune simulations indicated strong primary and secondary immune responses in the host. Therefore, our study identifies a new target that could expedite the design of effective therapeutics, and the designed vaccine should be validated. Future directions include an extensive molecular interaction analysis, in silico cloning, wet-lab experiments, and evaluation and comparison of the designed candidate as both a DNA vaccine and protein vaccine.

돼지와 양돈장 및 농장 관계자 간에 발생하는 항생제 내성 유전자 전파 조사를 위한 플라스미드 염기서열 분석 (Plasmid Sequence Data Analysis to Investigate Antibiotic Resistance Gene Transfer among Swine, Swine Farm and Their Owners)

  • 정유진;이선우;유정식;이동훈;운노타쯔야
    • 한국환경농학회지
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    • 제42권4호
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    • pp.269-278
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    • 2023
  • Antibiotics either kill or inhibit the growth of bacteria. However, antibiotic-resistant bacteria are difficult to treat with antibiotics. Infections caused by such bacteria often lead to severe diseases. Antibiotic resistance genes (ARG) can be horizontally transmitted across different bacterial species, necessitating a comprehensive understanding of how ARGs spread across various environments. In this study, we analyzed the plasmid sequences of 33 extended-spectrum beta-lactamases (ESBL) producing Escherichia coli isolated from pigs, farms, and their owners. We conducted an antibiotic susceptibility test (AST) with aztreonam and seven other antibiotics, as well as whole genome sequencing (WGS) of the strains using MinION. Our results demonstrated that the plasmids that did not harbor ARGs were mostly non-conjugative, whereas the plasmids that harbored ARGs were conjugative. The arrangement of these ARGs exhibited a pattern of organization featuring a series of ARG cassettes, some of which were identical across the isolates collected from different sources. Therefore, this study suggests that the sets of ARG cassettes on plasmids were mostly shared between pigs and their owners. Hence, enhanced surveillance of ARG should be implemented in farm environments to proactively mitigate the risk of antibiotic-resistant bacterial infections.