• Title/Summary/Keyword: TET protein

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Chemical Composition, Phenolic Concentration and In Vitro Gas Production Characteristics of Selected Acacia Fruits and Leaves

  • Abdulrazak, S.A.;Orden, E.A.;Ichinohe, T.;Fujihara, T.
    • Asian-Australasian Journal of Animal Sciences
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    • v.13 no.7
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    • pp.935-940
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    • 2000
  • The objective of this study was to evaluate the nutritive value of selected fruits (pods and seeds) and leaves of acacia tree species namely; Acacia nubica (nubica), Acacia tortilis (tortilis) and Acacia brevispica (brevispica), Acacia reficiens (reficiens) and Acacia senegal (senegal). A wide variability in chemical composition, polyphenolics and gas production was recorded. The crude protein (CP) ranged from 131 to 238 g/kg DM. Neutral detergent fiber (NDF), acid detergent fiber (ADF) and lignin (ADL) were higher in senegal and significantly different (p<0.05) from other species. The nitrogen bound to fiber tended to be higher in leaves than the fruits, ranging from 2.6 to 11.3 g/kg NDF and 1.6 to 3.2 g/kg ADF. The leaves of reficiens and senegal had higher concentrations of total extractable phenolics (TEPH), total extractable tannins (TET) and total condensed tannins (TCT), but lower in NDF, ADF and ADL than the fruits of nubica, tortilis and brevispica. Mineral concentrations varied among species; all were relatively poor in phosphorus, moderate in calcium and magnesium, and rich in microelements. A significant (p<0.05) variation in gas production after 12, 24, 48, 72 and 96 h was recorded between species. Nubica had the highest (p<0.05) rate of gas production (0.0925) while the highest potential gas production was recorded in tortilis. A strong negative correlation between TEPH and TET with gas production after 24, 48, 72 and 96 was established (r=-0.72 to -0.82). Crude protein and TCT correlated negatively but also weakly with gas production characteristics. Organic matter digestibility calculated from gas production after 48 h (OMD48) ranged between 465 g/kg DM in reficiens and 611 g/kg DM in tortilis. The results of this study indicate that acacia species have the potential to be used as feed supplements.

Cloning and Characterization of the Paraquat Resistance-Related Genes from Ochrobactrum anthropi JW-2 (Ochrobactrum anthropi JW-2 유래의 Paraquat 내성유전자 PqrA의 주변 유전자군 분석)

  • Bae Eun-Kyung;Lee Hyo-Shin;Won Sung-Hye;Lee Byung-Hyun
    • Microbiology and Biotechnology Letters
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    • v.34 no.1
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    • pp.15-22
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    • 2006
  • A 4,971 bp chromosomal DNA fragment containing the pqrA, paraquat resistance gene, was cloned from Ochrobactrum anthropi JW-2, and the complete nucleotide sequence was determined. Nucleotide and deduced amino acid sequences of the fragment revealed the presence of 4 complete ORFs (orf2, pqrA, orf3, orf4) and two incomplete ORFs(orf1, orf5). Orf1, pqrA, orf4 and orf5 exists at the direct strand but orf2 and orf3 exists at the reverse complementary strand. Orf1 which of incomplete sequences without start codon shares homology with ATP binding region of the response regulator receiver. Orf2 shares high homology with members of the tetR family of transcriptional repressor which have a helix-turn-helix (H-T-H) motif. Therefore, the orf2 is predicted as a transcriptional repressor of pqrA and is designated as pqrR2. Orf3 shares high homology with the members of the lysR family acting as a transcriptional activator which have both of a H-T-H motif at the N-terminal region and substrate binding domain at the C-terminal region. Therefore, the orf3 is predicted as a transcriptional activator of pqrA and is designated as pqrR1. Orf4 shows homology with the periplasmic substrate-binding protein of amino acid ABC transporter. Orf5 which of incomplete sequences without stop codon revealed the homology with the permeases protein of amino acid ABC transporter.

Ensiled Green Tea Waste as Partial Replacement for Soybean Meal and Alfalfa Hay in Lactating Cows

  • Kondo, Makoto;Nakano, Masashi;Kaneko, Akemi;Agata, Hirobumi;Kita, Kazumi;Yokota, Hiroomi
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.7
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    • pp.960-966
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    • 2004
  • The purpose of this study was to evaluate the effects of protein supplementation of green tea waste (GTW) on the performance of lactating cows. Another aim was to increase resource utilization and to eliminate any environmental negative impact from the tea waste. GTW from a beverage company was ensiled at a low pH (<4.0) and high acetic acid and lactic acid concentration, and it contained high crude protein (CP, 34.8%), total extractable tannins (TET, 9.2%) and condensed tannin (CT, 1.7%). Two experiments were conducted to investigate the palatability and performance in lactating cows fed GTW. In the palatability trial, three lactating cows were allocated to three dietary treatments in a 3$\times$3 Latin square design. The animals were offered a total mixed ration (TMR) including GTW at rates of 0, 2.5 and 5.0% on a dry matter (DM) basis. Total DM intake was not different among the treatments. In the performance trial, four lactating cows were used in a 2$\times$2 Latin square design with a 3 week sampling period. GTW was incorporated into TMR at a rate of 5.0% on a DM and 10.0% on a CP basis. Thus GTW replaced alfalfa hay and soybean meal at a level of 25.0% on a DM. DM and CP intake were not affected by the inclusion of GTW, whereas TET and CT intake were significantly increased (p<0.001). Milk production, milk composition and the efficiency of milk production were not altered by the GTW inclusion. Although ruminal pH and VFA, and blood urea nitrogen were not changed, ruminal $NH_{3}-N$ and plasma total cholesterol were relatively low in the GTW group, but not significantly different. The excretion of urinary purine derivatives and estimated MN supply were also not significantly affected by GTW treatment. It is therefore concluded that GTW can be used as a protein source without any detrimental effects on the performance of lactating cows.

Expression of the Recombinant Porcine GH Gene In Vitro Using Tetracycline Inducible Expression System (In Vitro에서의 Tetracycline Inducible Expression System에 의한 재조합 돼지 성장호르몬 유전자의 발현)

  • Kwon Mo Sun;Koo Bon Chul;Kim Teoan
    • Reproductive and Developmental Biology
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    • v.29 no.1
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    • pp.49-55
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    • 2005
  • We cloned cDNA of the PGH(porcine growth hormone) gene and constructed retrovirus vector designed to express PGH gene under the regulation of CMV (cytomegalovirus) promoter. To maximize the expression, WPRE(woodchuck hepatitis virus posttranscriptional regulatory element) sequence was placed at the downstream of the PGH gene. After infection with recombinant viruses, approximately 1×10/sup 6/ PFF(porcine fetal fibroblast) cells released PGH protein into the media as much as 1,400 ng. In a subsequent experiment, a modifications of the retrovirus vector was made to express the PGH gene in a teracycline-inducible manner. In PFF cells carrying these viral vector sequences, addition of doxycycline to the media resulted in 2∼6 fold increase in PGH synthesis. In the modified retrovirus vectors, the WPRE sequence also played a role in boosting the effect of the tetracycline induction. This result indicates that our tetracycline-inducible expression system might be a promising candidate in alleviating the complicate physiological problems caused by constitutive expression of the exogenous genes in the transgenic animals.

Ectopic Overexpression of Coiled-Coil Domain Containing 110 Delays G2/M Entry in U2-OS Cells

  • Lee, Sue Nyoung;Hong, Kyeong-Man;Seong, Yeon Sun;Kwak, Sahng-June
    • Development and Reproduction
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    • v.24 no.2
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    • pp.101-111
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    • 2020
  • Coiled-coil domain containing 110 (CCDC110, KM-HN-1) is a protein containing C-terminal coiled-coil domain (CCD) which was previously discovered as a member of the human cancer/testis antigen (CTA). In addition, CCDC110 has both nuclear localization signal sequence and the leucine zipper motif. Although the functional role of CCDC110 has yet to be fully identified, the mRNA expression levels of CCDC110 are known to be highly elevated in various cancer types including testis, implying its relevance to cancer pathogenesis. In this study, we first developed several monoclonal antibody (mAb) hybridoma clones targeting CCDC110 and further isolated clone by characterizing for its specificity using immunoblotting and immunoprecipitation approaches with basal parenchymal sperm cells in testis tissue. Next, using these mAbs, we showed that the Tet-inducible overexpression of CCDC110 protein delayed the entry of G2/M phase in U2-OS osteosarcoma cells. Based on these results, we propose that CCDC110 plays a crucial role in cell cycle progression.

Identification of Three Positive Regulators in the Geldanamycin PKS Gene Cluster of Streptomyces hygroscopicus JCM4427

  • Kim, Won-Cheol;Lee, Jung-Joon;Paik, Sang-Gi;Hong, Young-Soo
    • Journal of Microbiology and Biotechnology
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    • v.20 no.11
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    • pp.1484-1490
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    • 2010
  • In the Streptomyces hygroscopicus JCM4427 geldanamycin biosynthetic gene cluster, five putative regulatory genes were identified by protein homology searching. Among those genes, gel14, gel17, and gel19 are located downstream of polyketide synthase genes. Gel14 and Gel17 are members of the LAL family of transcriptional regulators, including an ATP/GTP-binding domain at the N-terminus and a DNA-binding helix-turn-helix domain at the C-terminus. Gel19 is a member of the TetR family of transcriptional regulators, which generally act to repress transcription. To verify the biological significance of the putative regulators in geldanamycin production, they were individually characterized by gene disruption, genetic complementation, and transcriptional analyses. All three genes were confirmed as positive regulators of geldanamycin production. Specifically, Gel17 and Gel19 are required for gel14 as well as gelA gene expression.

Protective Effect of Resveratrol on the Oxidative Stress-Induced Inhibition of Gap Junctional Intercellular Communication in HaCaT Keratinocytes

  • Lee, Jong-Chan;Lee, Sun-Mee;Kim, Ji-Hyun;Ahn, Soo-Mi;Lee, Byeong-Gon;Chang, Ih-Seoup
    • Biomolecules & Therapeutics
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    • v.11 no.4
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    • pp.224-231
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    • 2003
  • The aim of this study was to investigate the effect of resveratrol on the oxidative stress-induced inhibition of gap junctional intercellular communication in HaCaT keratinocytes. Anti-oxidative activity of resveratrol was measured by $\alpha,\alpha$-diphenyl-$\beta$-picrylhydrazyl assay and dichlorodihydrofluorescein diacetate oxidation assay. Gap junctional intercellular communication in HaCaT keratinocytes was assessed using the scrape loading/dye transfer technique. Western blots and reverse transcription-polymerase chain reaction were also analyzed for connexin 43 protein and mRNA expression, respectively. Resveratrol scavenged directly the stable $\alpha,\alpha$-diphenyl-$\beta$-picrylhydrazyl radical over a concentration range of 4 mg/ml ($78.2{\pm}2.7$% of control) to 500 mg/ml ($29.9{\pm}4.2$% of control) and decreased the intracellular reactive oxygen species induced by ultraviolet A (UVA) irradiation ($89.3{\pm}1.1$% of UVA group), ultraviolet B (UVB) irradiation ($70.9{\pm}1.7$% of UVB group) and 12-0-tet-radecanoylphorbol-13-acetate (TPA, $48.3{\pm}1.1$% of TPA group), respectively. UVA irradiation and TPA markedly reduced gap junctional intercellular communication, which was restored by resveratrol. There were no significant differences in the level of connexin 43 protein and mRNA expression among any of the experimental groups. Our data suggests that resveratrol has the protective effect on the oxidative stress-induced inhibition of gap junctional intercellular communication in HaCaT keratinocytes, and this protection is likely due to the scavenging of reactive oxygen species.

Virulence gene profiles and antimicrobial susceptibility of Salmonella Brancaster from chicken

  • Evie Khoo ;Roseliza Roslee ;Zunita Zakaria;Nur Indah Ahmad
    • Journal of Veterinary Science
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    • v.24 no.6
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    • pp.82.1-82.12
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    • 2023
  • Background: The current conventional serotyping based on antigen-antisera agglutination could not provide a better understanding of the potential pathogenicity of Salmonella enterica subsp. enterica serovar Brancaster. Surveillance data from Malaysian poultry farms indicated an increase in its presence over the years. Objective: This study aims to investigate the virulence determinants and antimicrobial resistance in S. Brancaster isolated from chickens in Malaysia. Methods: One hundred strains of archived S. Brancaster isolated from chicken cloacal swabs and raw chicken meat from 2017 to 2022 were studied. Two sets of multiplex polymerase chain reaction (PCR) were conducted to identify eight virulence genes associated with pathogenicity in Salmonella (invasion protein gene [invA], Salmonella invasion protein gene [sipB], Salmonella-induced filament gene [sifA], cytolethal-distending toxin B gene [cdtB], Salmonella iron transporter gene [sitC], Salmonella pathogenicity islands gene [spiA], Salmonella plasmid virulence gene [spvB], and inositol phosphate phosphatase gene [sopB]). Antimicrobial susceptibility assessment was conducted by disc diffusion method on nine selected antibiotics for the S. Brancaster isolates. S. Brancaster, with the phenotypic ACSSuT-resistance pattern (ampicillin, chloramphenicol, streptomycin, sulphonamides, and tetracycline), was subjected to PCR to detect the corresponding resistance gene(s). Results: Virulence genes detected in S. Brancaster in this study were invA, sitC, spiA, sipB, sopB, sifA, cdtB, and spvB. A total of 36 antibiogram patterns of S. Brancaster with a high level of multidrug resistance were observed, with ampicillin exhibiting the highest resistance. Over a third of the isolates displayed ACSSuT-resistance, and seven resistance genes (β-lactamase temoneira [blaTEM], florfenicol/chloramphenicol resistance gene [floR], streptomycin resistance gene [strA], aminoglycoside nucleotidyltransferase gene [ant(3")-Ia], sulfonamides resistance gene [sul-1, sul-2], and tetracycline resistance gene [tetA]) were detected. Conclusion: Multidrug-resistant S. Brancaster from chickens harbored an array of virulence-associated genes similar to other clinically significant and invasive non-typhoidal Salmonella serovars, placing it as another significant foodborne zoonosis.

Chemical Composition, Degradation Characteristics and Effect of Tannin on Digestibility of Some Browse Species from Kenya Harvested during the Wet Season

  • Osuga, I.M.;Abdulrazak, S.A.;Ichinohe, T.;Fujihara, T.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.1
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    • pp.54-60
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    • 2005
  • A study was conducted with the objective of evaluating the nutritive value of some browse species from Kenya. The species evaluated included: Bauhinia alba, Bauhinia variegata, Bridelia micrantha, Calliandra calothyrsus, Carisa edulis, Cratylia argentea, Gliricidia sepium, Lantana camara, Maerua angolensis, Sesbania micrantha and S. sesban. The browses were evaluated by their chemical composition including phenolics, in vitro gas production and tannin activity (tannin bioassay). All the species had high crude protein content (149-268 g/kg DM) and low NDF content (239-549 g/kg DM). The feeds had varying contents of total extractable tannins (TET) ranging from low (3-22 mg/g DM), moderate (42-58 mg/g DM) and high (77-152 mg/g DM). Calliandra calothyrsus had the highest tannin content. Significant (p<0.05) variation in gas production was recorded among the species. Sesbania micrantha had the highest (p<0.05) potential gas production while Gliricidia sepium had the highest (p<0.05) rate of gas production. Use of polyethylene glycol (PEG 6000), to assess the adverse affect of tannins, indicated that tannins in browse species with high tannin content had inhibitory effects on rumen microbial fermentation as indicated by the gas production. Estimated organic matter digestibility and metabolizable energy also increased with PEG addition. The results of this study indicate that such Kenyan browse species have the potential to be used as feed supplements for ruminant animals.

Novel Disease Model of Chronic Neutrophilic Leukemia: by Using the Tet-off System

  • Park, Jun-Hong;Lee, Young-Soon;Ryoo, Zae-Young
    • Proceedings of the Korean Society of Developmental Biology Conference
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    • 2003.10a
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    • pp.107-107
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    • 2003
  • The activation of protooncogenes or the inactivation of their gene products may be a specific and effective functional study for human neoplasia. To examine this possibility, we have used the tetracycline regulatory system to generate transgenic mice that conditionally express the HccR-2 protooncogene in vivo. The new human cervical cancer protooncogene (HccR-2) was detected from cervical cancer cell line. To elucidate its biological functions, we generated transgenic mice that expressed the HccR-2 gene. The sustained expression of the HccR-2 transgene culminated chronic neutrophilic leukemia (CNL). CNL is a rare chronic myeloproliferative disorder that presents as a sustained, mature neutrophilic leukocytosis with few or no circulating immature granulocytes, the absence of peripheral blood monocytosis, basophilia, or eosinophilia, and infiltration of neutrophils at the liver, spleen and kidney. Mice expressing the HccR-2 and tetracycline-transactivating protein (tTa) transgene were found to have altered myeloid development that was characterized by increased percentages of mature neutrophil and band form neutrophil in the peripheral blood, liver and spleen. Activation of the transgene causes CNL. In our model, expression of HccR-2 transgene mice was similar in many respects to the human CNL. This model will be valuable not only for investigating the biological properties of the HccR-2 and other protooncogenes in vivo but also for analyzing the mechanism involved in the progression of CNL.

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