• Title/Summary/Keyword: Reference Gene

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Subgenus classification of Accnthcmoebc by riboprinting (Riboprinting에 의한 가시아메바속의 분류)

  • 정동일;유학선
    • Parasites, Hosts and Diseases
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    • v.36 no.2
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    • pp.69-80
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    • 1998
  • Subgenus classification of Acanthcmoeba remains uncertain. Twenty-three reference strains of Acanthnmoeba including 18 (neo)type-strains were subjected for classification at the subgenus level by riboprinting, PCR/RFLP analysis of 185 rRNA gene (rDNA) . On the dendrogram reconstructed on the basis of riboprint analyses, two type- strains (A. astronwxis and A. tubinshi) of morphological group 1 diverged early from the other strains and were quite distinct from each other. Four type-strains of morphological group 3. A. culbertsoni, A. polestinensis, A. healyi were considered taxonomically valid, but A. pustulosn was regarded as an invalid synonym of A. pclestinensis. Strains of morphological group 2 were classified into 6 subgroups. Among them, A. giulni which has an intron in its 185 rDNA was the most divergent from the remaining strains. Acanthcmoebc ccstellanii Castellani, A. quinc Vil3, A. Iugdunensis L3a. A. poIyphage Jones, A. trinngularis SH621, and A. cqstellanii Ma strains belonged to a subgroup, A. castellanii complex. However, A. quinc and A. Lugnunensis were regarded as synonyms of A. ccstellanii. The Chang strain could be regarded as A. hatchetti. Acanthcmoebo nauritaniensis, A. niuionensis, A. paranivionensis could be considered as synonyms of A. rhwsodes. Neff strain was regarded as A. polyphage rather than as A. castellanii. It is likely that riboprinting can be applied for rapid identification of Acnnthcmoebc isolated from the clinical specimens and environments.

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Whole-Genome Resequencing Analysis of Hanwoo and Yanbian Cattle to Identify Genome-Wide SNPs and Signatures of Selection

  • Choi, Jung-Woo;Choi, Bong-Hwan;Lee, Seung-Hwan;Lee, Seung-Soo;Kim, Hyeong-Cheol;Yu, Dayeong;Chung, Won-Hyong;Lee, Kyung-Tai;Chai, Han-Ha;Cho, Yong-Min;Lim, Dajeong
    • Molecules and Cells
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    • v.38 no.5
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    • pp.466-473
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    • 2015
  • Over the last 30 years, Hanwoo has been selectively bred to improve economically important traits. Hanwoo is currently the representative Korean native beef cattle breed, and it is believed that it shared an ancestor with a Chinese breed, Yanbian cattle, until the last century. However, these two breeds have experienced different selection pressures during recent decades. Here, we whole-genome sequenced 10 animals each of Hanwoo and Yanbian cattle (20 total) using the Illumina HiSeq 2000 sequencer. A total of approximately 3.12 and 3.07 billion sequence reads were mapped to the bovine reference sequence assembly (UMD 3.1) at an average of approximately 10.71- and 10.53-fold coverage for Hanwoo and Yanbian cattle, respectively. A total of 17,936,399 single nucleotide polymorphisms (SNPs) were yielded, of which 22.3% were found to be novel. By annotating the SNPs, we further retrieved numerous nonsynonymous SNPs that may be associated with traits of interest in cattle. Furthermore, we performed whole-genome screening to detect signatures of selection throughout the genome. We located several promising selective sweeps that are potentially responsible for economically important traits in cattle; the PPP1R12A gene is an example of a gene that potentially affects intramuscular fat content. These discoveries provide valuable genomic information regarding potential genomic markers that could predict traits of interest for breeding programs of these cattle breeds.

Genetic Variation in the Immunoregulatory Gene of Adenovirus Type 3 (3형 아데노바이러스의 면역조절 유전자 다양성)

  • Choi, Eun Hwa;Kim, Hee Sup;Lee, Hoan Jong
    • Pediatric Infection and Vaccine
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    • v.16 no.2
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    • pp.199-204
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    • 2009
  • Purpose : Various proteins encoded in the early region 3 (E3) of adenoviruses protect cells from being killed by cytotoxic T cells and death-inducing cytokines. We sought to find out whether the genetic heterogeneity of the E3 gene might contribute to the molecular diversity of adenoviruses. Methods : Sequences in the E3 region were analyzed for 14 adenovirus type 3 (Ad3) strains that were isolated from children with lower respiratory tract infections in the Seoul National University Children's Hospital during the period 1991-2000. Full-length adenoviral DNA was purified from the infected A549 cell lysates using a modified Hirt procedure. Results : There was 98% homology between 14 Korean Ad3 strains with a reference strain (M15952). Homology within the Korean Ad3 strains was 98.7%. Variation was found in the region of transcripts 20.1 kDa, 20.6 kDa, truncated 7.7 kDa, 10.3 kDa, 14.9 kDa, and 15.3 kDa. In particular, all 14 Korean strains showed a missense single point mutation at the start codon of the truncated 7.7 kDa. In addition, a deletion was found in the truncated 7.7 kDa region by 58 base pairs in 10 strains and 94 base pairs in 4 strains. Variations in amino acids were observed in the receptor internalization and degradation complex (10.3 kDa/14.9 kDa) which stimulates the clearance from the cell surface and subsequent degradation of the receptors for the Fas ligand and TRAIL, while no variations were observed in another immunoregulatory transcript, 19 kDa. Conclusion : Sequence analysis of the immunoregulatory region of adenovirus E3 shows that genetic heterogeneities are related to genome type patterns.

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Diagnostic Evaluation of Non-Interpretable Results Associated with rpoB Gene in Genotype MTBDRplus Ver 2.0

  • Singh, Binit Kumar;Sharma, Rohini;Kodan, Parul;Soneja, Manish;Jorwal, Pankaj;Nischal, Neeraj;Biswas, Ashutosh;Sarin, Sanjay;Ramachandran, Ranjani;Wig, Naveet
    • Tuberculosis and Respiratory Diseases
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    • v.83 no.4
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    • pp.289-294
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    • 2020
  • Background: Line probe assay (LPA) is standard diagnostic tool to detect multidrug resistant tuberculosis. Non-interpretable (NI) results in LPA (complete missing or light wild-type 3 and 8 bands with no mutation band in rpoB gene region) poses a diagnostic challenge. Methods: Sputum samples obtained between October 2016 and July 2017 at the Intermediate Reference Laboratory, All India Institute of Medical Sciences Hospital, New Delhi, India were screened. Smear-positive and smear-negative culture-positive specimens were subjected to LPA Genotype MTBDRplus Ver 2.0. Smear-negative with culture-negative and culture contamination were excluded. LPA NI samples were subjected to phenotypic drug susceptibility testing (pDST) using MGIT-960 and sequencing. Results: A total of 1,614 sputum specimens were screened and 1,340 were included for the study (smear-positive [n=1,188] and smear-negative culture-positive [n=152]). LPA demonstrated 1,306 (97.5%) valid results with TUB (Mycobacterium tuberculosis) band, 24 (1.8%) NI, three (0.2%) valid results without TUB band, and seven (0.5%) invalid results. Among the NI results, 22 isolates (91.7%) were found to be rifampicin (RIF) resistant and two (8.3%) were RIF sensitive in the pDST. Sequencing revealed that rpoB mutations were noted in all 22 cases with RIF resistance, whereas the remaining two cases had wild-type strains. Of the 22 cases with rpoB mutations, the most frequent mutation was S531W (n=10, 45.5%), followed by S531F (n=6, 27.2%), L530P (n=2, 9.1%), A532V (n=2, 9.1%), and L533P (n=2, 9.1%). Conclusion: The present study showed that the results of the Genotype MTBDRplus assay were NI in a small proportion of isolates. pDST and rpoB sequencing were useful in elucidating the cause and clinical meaning of the NI results.

Genome-wide association study for the interaction between BMR and BMI in obese Korean women including overweight

  • Lee, Myoungsook;Kwon, Dae Young;Kim, Myung-Sunny;Choi, Chong Ran;Park, Mi-Young;Kim, Ae-jung
    • Nutrition Research and Practice
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    • v.10 no.1
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    • pp.115-124
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    • 2016
  • BACKGROUND/OBJECTIVES: This is the first study to identify common genetic factors associated with the basal metabolic rate (BMR) and body mass index (BMI) in obese Korean women including overweight. This will be a basic study for future research of obese gene-BMR interaction. SUBJECTS/METHODS: The experimental design was 2 by 2 with variables of BMR and BMI. A genome-wide association study (GWAS) of single nucleotide polymorphisms (SNPs) was conducted in the overweight and obesity (BMI > $23kg/m^2$) compared to the normality, and in women with low BMR (< 1426.3 kcal/day) compared to high BMR. A total of 140 SNPs reached formal genome-wide statistical significance in this study (P < $1{\times}10^{-4}$). Surveys to estimate energy intake using 24-h recall method for three days and questionnaires for family history, a medical examination, and physical activities were conducted. RESULTS: We found that two NRG3 gene SNPs in the 10q23.1 chromosomal region were highly associated with BMR (rs10786764; $P=8.0{\times}10^{-7}$, rs1040675; $2.3{\times}10^{-6}$) and BMI (rs10786764; $P=2.5{\times}10^{-5}$, rs10786764; $6.57{\times}10^{-5}$). The other genes related to BMI (HSD52, TMA16, MARCH1, NRG1, NRXN3, and STK4) yielded P < $10{\times}10^{-4}$. Five new loci associated with BMR and BMI, including NRG3, OR8U8, BCL2L2-PABPN1, PABPN1, and SLC22A17 were identified in obese Korean women (P < $1{\times}10^{-4}$). In the questionnaire investigation, significant differences were found in the number of starvation periods per week, family history of stomach cancer, coffee intake, and trial of weight control in each group. CONCLUSION: We discovered several common BMR- and BMI-related genes using GWAS. Although most of these newly established loci were not previously associated with obesity, they may provide new insights into body weight regulation. Our findings of five common genes associated with BMR and BMI in Koreans will serve as a reference for replication and validation of future studies on the metabolic rate.

Development of SNP Molecular Marker for Red-fleshed Color Identification of Peach Genetic Resources (복숭아 유전자원의 적색 과육 판별 SNP 분자표지 개발)

  • Kim, Se Hee;Nam, Eun Young;Cho, Kang Hee;Jun, Ji Hae;Chung, Kyeong Ho
    • Korean Journal of Plant Resources
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    • v.32 no.4
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    • pp.303-311
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    • 2019
  • Various colors of fruit skin and flesh are the most popular commercial criteria for peach classification. In order to breed new red-fleshed peach cultivar, many cross seedlings and generations should be maintained. Therefore it is necessary to develop early selection markers to screen seedlings with target traits to increase breeding efficiency. For the comparison of transcription profiles in peach cultivars differing in flesh color expression, two cDNA libraries were constructed. Differences in gene expression between red-fleshed peach cultivar, 'Josanghyeoldo' and white-fleshed peach cultivar, 'Mibaekdo' were analyzed by next-generation sequencing (NGS). Expressed sequence tag (EST) of clones from the two cultivars were selected for nucleotide sequence determination and homology searches. Putative single nucleotide polymorphisms (SNP) were screened from peach EST contigs by high resolution melting (HRM) analysis displayed specific difference between 8 red-fleshed peach cultivars and 24 white-fleshed peach cultivars. All 72 pairs of SNPs were discriminated and the HRM profiles of amplicons were established. In the study reported here, the development of SNP markers for distinguishing between red and white fleshed peach cultivars by HRM analysis offers the opportunity to use DNA markers. This SNP marker could be useful for peach marker assisted breeding and provide a good reference for relevant research on molecular mechanisms of color variation in peach cultivars.

Current Status and Prospect of Wheat Functional Genomics using Next Generation Sequencing (차세대 염기서열분석을 통한 밀 기능유전체 연구의 현황과 전망)

  • Choi, Changhyun;Yoon, Young-Mi;Son, Jae-Han;Cho, Seong-Woo;Kang, Chon-Sik
    • Korean Journal of Breeding Science
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    • v.50 no.4
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    • pp.364-377
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    • 2018
  • Hexaploid wheat (common wheat/bread wheat) is one of the most important cereal crops in the world and a model for research of an allopolyploid plant with a large, highly repetitive genome. In the heritability of agronomic traits, variation in gene presence/absence plays an important role. However, there have been relatively few studies on the variation in gene presence/absence in crop species, including common wheat. Recently, a reference genome sequence of common wheat has been fully annotated and published. In addition, advanced next-generation sequencing (NGS) technology provides high quality genome sequences with continually decreasing NGS prices, thereby dawning full-scale wheat functional genomic studies in other crops as well as common wheat, in spite of their large and complex genomes. In this review, we provide information about the available tools and methodologies for wheat functional genomics research supported by NGS technology. The use of the NGS and functional genomics technology is expected to be a powerful strategy to select elite lines for a number of germplasms.

SNP Markers Useful for the Selection of Yellow-fleshed Peach Cultivar (황육계 복숭아 품종 선발용 SNP 마커)

  • Kim, Se Hee;Kwon, Jung-hyun;Cho, Kang Hee;Shin, Il Sheob;Jun, Ji Hae;Cho, Sang-Yun
    • Korean Journal of Plant Resources
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    • v.34 no.5
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    • pp.443-450
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    • 2021
  • Peach flesh color is commercially important criteria for classification and has implications for nutritional quality. To breed new yellow-fleshed peach cultivar many cross seedlings and generations should be maintained. Therefore it is necessary to develop early selection molecular markers for screening cross seedlings and germplasm with economically important traits to increase breeding efficiency. For the comparison of transcription profiles in peach varieties with a different flesh color expression, two cDNA libraries were constructed. Differences in gene expression between yellow-fleshed peach cultivar, 'Changhowon Hwangdo' and white-fleshed peach cultivar, 'Mibaekdo' were analyzed by next-generation sequencing (NGS). Expressed sequence tag (EST) of clones from the two varieties was selected for nucleotide sequence determination and homology searches. Putative single nucleotide polymorphisms (SNPs) were screened from peach EST contigs by high resolution melting (HRM) analysis, SNP ID ppa002847m:cds and ppa002540m:cds displayed specific difference between 17 yellow-fleshed and 21 white-fleshed peach varieties. The SNP markers for distinguishing yellow and white fleshed peach varieties by HRM analysis offers the opportunity to use early selection. This SNP markers could be useful for marker assisted breeding and provide a good reference for relevant research on molecular mechanisms of color variation in peach varieties.

Food Fraud Monitoring of Commercial Sciaenidae Seafood Product Using DNA Barcode Information (DNA barcode를 이용한 민어과 수산가공품 진위판별 모니터링)

  • Park, Eun-Ji;Jo, Ah-Hyeon;Kang, Ju-Yeong;Lee, Han-Cheol;Park, Min-Ji;Yang, Ji-Young;Shin, Ji-Young;Kim, Gun-Do;Kim, Jong-Oh;Seo, Yong-Bae;Kim, Jung-Beom
    • Journal of Food Hygiene and Safety
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    • v.35 no.6
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    • pp.574-580
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    • 2020
  • In this study we sought to determine the food fraud by discriminating species of commercial seafood product such as Larimichthys polyactis, Larimichthys crocea, Pennahia argentatus, and Miichthys miiuy, which are difficult to morphologically discriminate. After amplifying the mitochondrial cytochrome c oxidase subunit I gene of the reference fish, the DNA sequences of the amplified PCR products were analyzed. As a result, a 655 bp sequence for species identification was selected for use as DNA barcodes. To confirm the DNA data and primer set, the DNA barcode sequence of each fish was compared to that in that in the NCBI. All of the DNA barcode data were matched with the gene sequence of each fish in the NCBI. A total of 32 processed seafood products (8 L. polyactis, 12 L. crocea, 3 Pennahia argentatus, and 9 Miichthys miiuy) were investigated. Homology of 97% or more in DNA sequences was judged as the same species. As a result of the monitoring, there were no discovered cases of forgery or alteration. However, the use of a raw material name having no matching standard name in the Korea Food Code may cause consumer confusion. Therefore, it is suggested that the standard name or scientific name be co-labeled with the raw material name on seafood products to prevent consumer confusion.

Genetic diversity and selection of Tibetan sheep breeds revealed by whole-genome resequencing

  • Dehong Tian;Buying Han;Xue Li;Dehui Liu;Baicheng Zhou;Chunchuan Zhao;Nan Zhang;Lei Wang;Quanbang Pei;Kai Zhao
    • Animal Bioscience
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    • v.36 no.7
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    • pp.991-1002
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    • 2023
  • Objective: This study aimed to elucidate the underlying gene regions responsible for productive, phenotypic or adaptive traits in different ecological types of Tibetan sheep and the discovery of important genes encoding valuable traits. Methods: We used whole-genome resequencing to explore the genetic relationships, phylogenetic tree, and population genetic structure analysis. In addition, we identified 28 representative Tibetan sheep single-nucleotide polymorphisms (SNPs) and genomic selective sweep regions with different traits in Tibetan sheep by fixation index (Fst) and the nucleotide diversity (θπ) ratio. Results: The genetic relationships analysis showed that each breed partitioned into its own clades and had close genetic relationships. We also identified many potential breed-specific selective sweep regions, including genes associated with hypoxic adaptability (MTOR, TRHDE, PDK1, PTPN9, TMTC2, SOX9, EPAS1, PDGFD, SOCS3, TGFBR3), coat color (MITF, MC1R, ERCC2, TCF25, ITCH, TYR, RALY, KIT), wool traits (COL4A2, ERC2, NOTCH2, ROCK1, FGF5, SOX9), and horn phenotypes (RXFP2). In particular, a horn-related gene, RXFP2, showed the four most significantly associated SNP loci (g. 29481646 A>G, g. 29469024 T>C, g. 29462010 C>T, g. 29461968 C>T) and haplotypes. Conclusion: This finding demonstrates the potential for genetic markers in future molecular breeding programs to improve selection for horn phenotypes. The results will facilitate the understanding of the genetic basis of production and adaptive unique traits in Chinese indigenous Tibetan sheep taxa and offer a reference for the molecular breeding of Tibetan sheep.