• Title/Summary/Keyword: supF gene

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Genome-based identification of strain KCOM 1265 isolated from subgingival plaque at the species level

  • Park, Soon-Nang;Lim, Yun Kyong;Kook, Joong-Ki
    • International Journal of Oral Biology
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    • v.45 no.2
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    • pp.70-75
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    • 2020
  • The aim of this study was to identify strain KCOM 1265 isolated from subgingival plaque at the species level by comparing 16S ribosomal RNA gene (16S rDNA) and genome sequences. The whole genome of strain KCOM 1265 was extracted using the phenol-chloroform extraction method. 16S rDNA was amplified using polymerase chain reaction and sequenced using the dideoxy chain termination method. Pairwise genome comparison was performed using average nucleotide identity (ANI) and genome-to-genome distance (GGD) analyses. The data showed that the percent similarity of 16S rDNA sequence of strain KCOM 1265 was 99.6% as compared with those of Fusobacterium polymorphum ATCC 10953T and Fusobacterium hwasookii KCOM 1249T. The ANI values of strain KCOM 1265 with F. polymorphum ATCC 10953T and F. hwasookii KCOM 1249T were 95.8% and 93.0%, respectively. The GGD values of strain KCOM 1265 with F. polymorphum ATCC 10953T and F. hwasookii KCOM 1249T were 63.9% and 49.6%, respectively. These results indicate that strain KCOM 1265 belongs to F. polymorphum.

Development of strain-specific polymerase chain reaction primers to detect Fusobacterium hwasookii strains

  • Lim, Yun Kyong;Kook, Joong-Ki
    • International Journal of Oral Biology
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    • v.46 no.4
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    • pp.155-159
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    • 2021
  • This study aimed to develop strain-specific polymerase chain reaction (PCR) primers to detect Fusobacterium hwasookii KCOM 1249T, F. hwasookii KCOM 1253, F. hwasookii KCOM 1256, F. hwasookii KCOM 1258, and F. hwasookii KCOM 1268 on the basis of nucleotide sequences of a gene specific to each strain. The unique genes for each F. hwasookii strain were determined on the basis of their genome sequences using Roary. The strain-specific PCR primers based on each strain-specific gene were designed using PrimerSelect. The specificity of each PCR primer was determined using the genomic DNA of the 5 F. hwasookii strains and 25 strains of oral bacterial species. The detection limit and sensitivity of each strain-specific PCR primer pair were determined using the genomic DNA of each target strain. The results showed that the strain-specific PCR primers correspond to F. hwasookii KCOM 1249T, F. hwasookii KCOM 1253, F. hwasookii KCOM 1258, F. hwasookii KCOM 1256/F. nucleatum subsp. polymorphum KCOM 1260, or F. hwasookii KCOM 1268/Fusobacterium sp. oral taxon 203 were developed. The detection limits of these strain-specific PCR primers ranged from 0.2 to 2 ng of genomic DNA for each target strain. The results suggest that these strain-specific PCR primers are valuable in quality control for detecting specific F. hwasookii strains.

Association between Genotypes of the Isocitrate Dehydrogenase 3, beta subunit (IDH3B) Gene and Carcass Traits in an F2 Crossbred Population of Landrace × Jeju (Korea) Black Pigs (제주흑돼지와 랜드레이스 교배 2세대의 도체형질과 IDH3B 유전자형의 상관관계)

  • Kang, Yong-Jun;Jeong, Dong Kee;Cho, In-Cheol;Han, Sang-Hyun
    • Journal of Life Science
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    • v.26 no.4
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    • pp.414-418
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    • 2016
  • This study tested the association between genetic polymorphisms of the isocitrate dehydrogenase 3, beta subunit (IDH3B) gene and economic traits in an F2 crossbred population of Landrace × Jeju (South Korea) Black pigs. A 304-bp insertion/deletion mutation in promoter region was screened for determining genotypes of the IDH3B gene in a total of 1,105 F2 pigs. Three genotypes (AA, AB, and BB) were identified in the founder, F1, and F2 populations. Association analysis showed significant differences in carcass weights (CW), backfat thicknesses in three positions of the body (4th-5th ribs, BF5; 11th-12th ribs, BF12; 13th rib-1st lumbar, BFL), and carcass lengths (CL) (p<0.05), but not in meat color (MC), eye muscle area (EMA), or marbling scores (MARB) (p>0.05). The F2 IDH3B BB homozygotes showed heavier CW (80.790±0.725 kg) and shorter CL (101.875±0.336 cm) than the other genotypes (p<0.05). In addition, the BF levels between the 4th - 5th and 11th - 12th vertebrae were thicker in the carcasses of pigs with the IDH3B BB genotype than with the other genotypes (p<0.05). These results suggested that genetic variations in the IDH3B gene may serve as molecular genetic markers for improving the Landrace × Jeju Black pig crossbreeding systems.

Comparison of D-[18F]FMAU and L-[18F]FMAU as PET Imaging Agents for HSV1-TK Gene Expression

  • Moon, Byung-Seok;Jo, Nam-Hyun;Lee, Kyo-Chul;El-Gamal, Mohammed I.;An, Gwang-Il;Hong, Su-Hee;Choi, Tae-Hyun;Choi, Won-Kyoung;Park, Jin-Hun;Cho, Jung-Hyuck;Cheon, Gi-Jeong;Oh, Chang-Hyun
    • Bulletin of the Korean Chemical Society
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    • v.31 no.11
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    • pp.3309-3312
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    • 2010
  • D-[$^{18}F$]FMAU and L-[$^{18}F$]FMAU are F-18 labeled nucleoside analogue which have been efficiently synthesized in order to be a PET imaging probe. D-[$^{18}F$]FMAU and L-[$^{18}F$]FMAU were compared as PET imaging agents using HSV1-TK gene expressing tumor-bearing mice. Their cellular uptake profiles were also compared using MCA and MCA-TK cell lines. D-[$^{18}F$]FMAU demonstrated higher cellular uptake and higher accumulation in MCA-TK tumor regions than L-[$^{18}F$]FMAU. On the other hand, L-[$^{18}F$]FMAU showed higher MCA-TK/MCA ratio of %ID/g than that of D-[$^{18}F$]FMAU. L-[$^{18}F$]FMAU can be utilized as a good candidate for HSV1-TK PET imaging. It can be used for antiviral drug evaluation.

Study on Antitumor Activity and Immunomodulatory effects of Seoleosojong-tang (활어소종탕이 항종역반응에 미치는 영향)

  • Son Ki Jeong;Park Yang Chun
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.18 no.1
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    • pp.137-147
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    • 2004
  • In order to evaluate the antitumor activity and immunomodulatory effects of Seoleosojong-tang(SST), studies were done. We measured the cytotoxic activity for various kinds of cancer cells, inhibitory effect on activity of DNA topoisomerase I, cell adhesion to complex extracellular matrix, survival time in ICR bearing S-180, pulmonary colonization and histological changes of lung in C57BL/6 injected i.v. with B16-F10, CAM assay, expression of CD4/sup +/, CD8/sup +/, B220/sup +/, cytokine gene in spleen cell. The results were obtained as follows: 1. In cytotoxicity against A549, HT1080, 816-F10, NCL-H661 was showed cytotoxicity as compared with control. 2. The inhibitory effect on adhesion of A549, 816-F10 to complex extracellular matrix was over 40% at 100 ㎍/㎖ of SST. 3. In DNA topoisomerase I assay, SST has inhibitory effect. 4. The T/C% was 120.8 in SST treated group in S-180 bearing ICR mice. 5. In pulmonary colonization assay, a number of colonies were decreased significantly and histological changes were showed that infiltration area of cancer cells were inhibited effectively in SST treated group. 6. In CAM Assay, SST has antiangiogenic effect. 7. On the expression of positive cell to CD4/sup +/, CD8/sup +/ and 8220/sup +/ in spleen cells, CD4/sup +/ cells were increased significantly in SST treated group. 8. Effect of SST on IL-1β gene expression in splenic cell was significantly increased as function of whole concentration. 9. The gene expression of IL-4, IL-6, IL-10, IL-12, IFN-γ, TNF-α were increased in SST treated group. From above results SST could be usefully applied for antitumor activity and immunomodulatory effects, but further research of SST should be required.

Isolation and characterization of feline endometrial mesenchymal stem cells

  • Mi-Kyung Park;Kun-Ho Song
    • Journal of Veterinary Science
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    • v.25 no.2
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    • pp.31.1-31.8
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    • 2024
  • Background: Recently, there has been a growing interest in stem cells for human medicine. Limited feline endometrial mesenchymal stem cell (fEM-MSC) research in veterinary medicine necessitates reporting for future feline disease research and therapy. Objectives: This study aimed to isolate fEM-MSCs from feline endometrial tissues and evaluate their morphology, proliferative ability, differentiation ability, and immunophenotype. Methods: Feline endometrial tissues were obtained from the ovariohysterectomies of healthy cats and isolated using an enzymatic method. The morphology and proliferative ability of the isolated cells were assessed using a doubling time (DT) assay from passages 3 to 6 (P3 - P6). We measured pluripotency gene expressions of cells in P2 using quantitative real-time polymerase chain reaction (qRT-PCR). To investigate MSC characteristics, a trilineage differentiation assay was conducted in P4, and cells in P4 were immunophenotyped using flow cytometry. Results: fEM-MSCs showed a typical spindle-shaped morphology under a microscope, and the DT was maintained from P3 to P6. fEM-MSCs could differentiate into adipocytes, osteoblasts, and chondrocytes, and expressed three pluripotency markers (OCT4, SOX2, and NANOG) by qRT-PCR. Immunophenotypic analysis showed that the fEM-MSCs were CD14 -, CD34 -, CD45 -, CD9+, and CD44+. Conclusions: In this study, the feline endometrium was a novel source of MSCs, and to the best of our knowledge, this is the first report on the isolation method and characteristics of fEM-MSCs.

ON THE ADAPTED EQUATIONS IN VARIOUS DYPLOID MODEL AND HARDY-WEINBURG EQUILIBRIUM IN A TRIPLOID MODEL

  • Won Choi
    • Korean Journal of Mathematics
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    • v.31 no.1
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    • pp.17-23
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    • 2023
  • For a locus with two alleles (IA and IB), the frequencies of the alleles are represented by $$p=f(I^A)={\frac{2N_{AA}+N_{AB}}{2N}},\;q=f(I^B)={\frac{2N_{BB}+N_{AB}}{2N}}$$ where NAA, NAB and NBB are the numbers of IAIA, IAIB and IBIB respectively and N is the total number of populations. The frequencies of the genotypes expected are calculated by using p2, 2pq and q2. Choi defined the density and operator for the value of the frequency of one gene and found the adapted partial differential equation as a follow-up for the frequency of alleles and applied this adapted partial differential equation to several diploid model [1]. In this paper, we find adapted equations for the model for selection against recessive homozygotes and in case that the alley frequency changes after one generation of selection when there is no dominance. Also we consider the triploid model with three alleles IA, IB and i and determine whether six genotypes observed are in Hardy-Weinburg for equilibrium.

Improvement of Pre-harvest Sprouting Resistance in Korean japonica Varieties through a Precision Marker-based Breeding

  • Kamal Bhattarai;Patricia Izabelle Lopez;Sherry Lou Hechanova;Ji-Ung Jeung;Hyun-Sook Lee;Eok-Keun Ahn;Ung-Jo Hyun;Jong-Hee Lee;So-Myeong Lee;Jose E. Hernandez;Sung-Ryul Kim
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2022.10a
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    • pp.269-269
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    • 2022
  • Pre-harvest sprouting (PHS) on rice panicles is getting problematic in recent several years in Korea due to climate changes such as high temperature and more frequent typhoons during harvesting season. PHS negatively affects grain quality severely and also yield. Genetic improvement of Korean varieties (Oryza sativa ssp. japonica) through a marker assisted-backcross breeding (MAB) with the known PHS resistant genes must be one of ideal solutions. However, the final breeding products of MAB occasionally exhibit unwanted traits, especially the cross between genetically distant parents. This might be caused by linkage drag and/or presence of the gene-unlinked donor introgressions, resulting that the final products could not be released to the farmers. The major PHS resistance gene, Sdr4 (Seed dormancy 4) originated from an indica cultivar, Kasalath was selected as a donor gene. In order to avoid unexpected phenotypes in the breeding products, we performed a precision marker-based breeding (PMBB) consisting of foreground, recombinant, and background selections (FS, RS, and BS) which aim to develop 'single small introgression lines' (~100 kb introgression). Korean varieties (Ilpum and Gopum) were crossed with Kasalath. We developed Sdr4-allele specific markers for FS and a set of polymorphic flanking markers near the Sdr4 (-350kb and +420kb) for RS. To minimize linkage drag, the small introgression (< 125kb) containing Sdr4 was selected in Ilpum background (BC2F4) through 1st RS with ~1,200 F2 or BC1F2 plants (one side trimmed) and then 2nd RS with ~1,000 progenies from the 1st RS selected plants (another side trimmed). After RS, the selected lines were genotyped by using Infinium 7K SNP chip to detect other donor introgressions and the lines were backcrossed. Currently BS is on-going from the backcross-derived progenies with BS markers to remove residual introgressions. During the PMBB process, genetic effect of Sdr-4-Kasalath allele was confirmed in Ilpum and Gopum backgrounds by PHS phenotyping using the segregating BC2F3 or BC1F4 materials. The Sdr4 PMBB lines in Ilpum background (< 125kb introgression) will be valuable genetic resources to improve PHS resistance in modem popular temperate japonica varieties.

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Characterization of a gene encoding ornithine carbamoyltransferase from rice

  • Islam Sikdar, Shafiqul;Kim, Jung-Sup
    • Journal of Plant Biotechnology
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    • v.36 no.4
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    • pp.397-402
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    • 2009
  • Ornithinine carbamoyltransferase (OTC) is an enzyme that catalyzes the key step in arginine biosynthesis in bacteria and plants. OTC is also involved in the urea cycle and deficiency of the enzyme in human leads to disease. The argF gene encoding OTC has been reported in many bacteria and few plants. Here we report the characterization of a gene encoding OTC from rice (OsOTC). Analysis of a cDNA sequence from rice revealed that the full-length open reading frame of OsOTC consisted of 367 amino acids, corresponding to a protein of approximately 39.7 kDa. The predicted amino acid sequence of OsOTC harbor distinct five OTC signature sites and is highly homologous to that of enzymes of plants, animals and many bacterial OTCs. Expression of OsOTC in argF mutants of Escherichia coli showed that the gene was able to functionally complement to the mutant. These results suggest that the OsOTC encode a protein for ornithine carbamoyltransferase in rice.

Mutation spectra induced by 1-nitropyrene 4,5-oxide and 1-nitropyrene 9,10-oxide in the supF gene of human XP-A fibroblasts

  • Kim, Byung-Wook;Kim, Byung-Chun;Cha, Jin-Soon;Pfeifer, Gerd P.;Lee, Chong-Soon
    • BMB Reports
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    • v.41 no.8
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    • pp.604-608
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    • 2008
  • 1-Nitropyrene 4,5-oxide and 1-nitropyrene 9,10-oxide are oxidative metabolites that are responsible for the mutagenicity of 1-nitropyrene. In this study, the mutation spectra induced by oxidative metabolites in human cells were determined using a shuttle vector assay. The mutation frequencies induced by 1-nitropyrene 9,10-oxide were 2-3 times higher than those induced by 1-nitropyrene 4,5-oxide. The base substitutions induced by 1-nitropyrene 4,5-oxide were $G{\rightarrow}A$ transitions, $G{\rightarrow}C$ transversions, and $G{\rightarrow}T$ transversions. In the case of 1-nitropyrene 9,10-oxide, $G{\rightarrow}A$ transitions, $G{\rightarrow}T$ transversions, $A{\rightarrow}G$ transitions and $G{\rightarrow}C$ transversions were observed. Most base substitution mutations induced by oxidative metabolites occurred at the guanine sites in the supF gene. These sequence-specific hot spots were commonly identified as 5'-GA sequences for both metabolites. On the other hand, the sequence-specific hot spots at the adenine sites were identified as 5'-CAC sequences for 1-nitropyrene 9,10-oxide. These results suggest that the oxidative metabolites of 1-nitropyrene induce sequence-specific DNA mutations at the guanine and adenine sites at high frequency.