• Title/Summary/Keyword: j-deletion

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Cloning of Chicken Microsomal Glutathione S-transferase 1 Gene (MGST1) and Identification of Its Different Splice Variants

  • Wang, X.-T.;Zhang, H.;Zhao, C.-J.;Li, J.-Y.;Xu, G.-Y.;Lian, L.-S.;Wu, C.-X.;Deng, Xuemei
    • Asian-Australasian Journal of Animal Sciences
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    • v.22 no.2
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    • pp.155-161
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    • 2009
  • Mammal microsomal glutathione transferase 1 (MGST1) can conjugate many toxic or carcinogenic substances and depress oxidative stress. In this study, Chicken MGST1 and its variants were cloned for the first time and were composed of 956 or 944 nucleotides. The 12 nt deletion in the exon 2 did not alter the GT-AG rule and the ORFs for the two MGST1 variants were the same, which both comprised 465 nucletides and encoded a peptide with 155 amino acids. It was found that the two different splice variants identified using RT-PCR expressed in all three organs investigated of Dwarf Brown Chicken, namely liver, spleen and shell gland. Moreover, the expression level of MGST1 mRNA in the liver of Dwarf Brown chickens was the highest (p<0.01), and there were no significant differences between the spleen and the shell gland. These results provide a base for studying the biological function of Chicken MGST1.

Complete Mitochondrial Genome Sequence and Genetic Diversity of Duroc Breed (돼지 Duroc 품종에서 미토콘드리아 유전체 서열의 특성과 집단의 유전적 다양성)

  • Cho, 1.C.;Han, S.H.;Choi, Y.L.;Ko, M.S.;Lee, J.G;Lee, J.H;Jeon, J .T
    • Journal of Animal Science and Technology
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    • v.46 no.6
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    • pp.937-946
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    • 2004
  • Duroc is widely used to improve the meat quality and productivity. To elucidate the phylogenetic relation and the sequence specificity for the maternal property, the complete sequence of mitochondrial genome was determined and the population diversity of Duroc was investigated in this study. The length of mtDNA tested is 16,584-bp. There are several insertion/deletion mutations in the control region and coding regions for tRNA and rRNA, respectively, but not in peptide-coding regions. Four peptide-coding genes(COⅡ, COⅢ, ND3 and ND4) showed incomplete termination codon sequences such as T--, and two(ND2 and ND4L) did alternative initiation codons(AIC), respectively. Especially, the initiation codon sequences of ND2 gene were polymorphic in this population. Polymorphisms were detected in 11-bp duplication motif within control region as well as ND2 and CYTB. Variation patterns observed from the tests on three mtDNA regions were linked completely and then two haplotypes obtained from combining the data dividing this population. Duroc mtDNA is observed at the European pig cluster in the phylogenetic tree, however, the results from the population analyses supported previous opinions. This study suggests that the breed Duroc was mainly originated from the European pig lineage, and Asian lineage was also used to form the pig breed Duroc as maternal progenitors.

Identification of a Promoter Motif Involved in Curtovirus Sense-Gene Expression in Transgenic Arabidopsis

  • Hur, Jingyung;Choi, Eunseok;Buckley, Kenneth J.;Lee, Sukchan;Davis, Keith R.
    • Molecules and Cells
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    • v.26 no.2
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    • pp.131-139
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    • 2008
  • Expression of the seven open reading frames (ORFs) of single-stranded DNA Curtoviruses such as Beet curly top virus (BCTV) and Beet severe curly top virus (BSCTV) is driven by a bi-directional promoter. To investigate this bidirectional promoter activity with respect to viral late gene expression, transgenic Arabidopsis plants expressing a GUS reporter gene under the control of either the BCTV or BSCTV bi-directional promoter were constructed. Transgenic plants harboring constructs showed higher expression levels when the promoter of the less virulent BCTV was used than when the promoter of the more virulent BSCTV was used. In transgenic seedlings, the reporter gene constructs were expressed primarily in actively dividing tissues such as root tips and apical meristems. As the transgenic plants matured, reporter gene expression diminished but viral infection of mature transgenic plants restored reporter gene expression, particularly in transgenic plants containing BCTV virion-sense gene promoter constructs. A 30 base pair conserved late element (CLE) motif was identified that was present three times in tandem in the BCTV promoter and once in that of BSCTV. Progressive deletion of these repeats from the BCTV promoter resulted in decreased reporter gene expression, but BSCTV promoters in which one or two extra copies of this motif were inserted did not exhibit increased late gene promoter activity. These results demonstrate that Curtovirus late gene expression by virion-sense promoters depends on the developmental stage of the host plant as well as on the number of CLE motifs present in the promoter.

Immune Tolerance in Murine Islet Transplantation Across HY Disparity (HY 항원 불일치 췌도 이식에 의한 면역 관용의 유도)

  • Choi, Seung-Eun;Park, Chung-Gyu
    • IMMUNE NETWORK
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    • v.4 no.1
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    • pp.53-59
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    • 2004
  • Background: Minor histocompatibility HY antigen, as a transplantation antigen, has been known to cause graft rejection in MHC (major histocompatibility complex) matched donor-recipient. The aim of our study is to investigate the role of male antigen (HY) disparity on MHC matched pancreatic islet transplantation and to examine the mechanism of the immune reaction. Methods: Pancreatic islets were isolated and purified by collagen digestion followed by Ficoll gradient. The isolated islets of male C57BL6/J were transplanted underneath the kidney capsule of syngeneic female mice rendered diabetic with streptozotocine. Blood glucose was monitored for the rejection of engrafted islets. After certain period of time, tail to flank skin transplantation was performed either on mouse transplanted with HY mismatched islets or on sham treated mouse. The rejection was monitored by scoring gross pathology of the engrafted skin. Results: HY mismatched islets survived more than 300 days in 14 out of 15 mice. The acceptance of second party graft (male B6 islets) and the rejection of third party graft (male BALB/c islets) in these mice suggested the tolerance to islets with HY disparity. B6 Skin with HY disparity was rejected on day $25{\pm}7$. However, HY mismatched skin transplanted on the mice tolerated to HY mismatched islets survived more than 240 days. Tetramer staining in these mice indicated the CTL recognizing MHC Db/Uty was not deleted or anergized. Conclusion: The islet transplantation across HY disparity induced tolerance to HY antigen in C57BL6 mouse, which in turn induced tolerance to HY mismatched skin, which otherwise would be rejected within 25 days. The MHC tetramer staining suggested the underlying mechanisms would not be clonal deletion or anergy.

Korean red ginseng attenuates HIV-1 vivo; High frequency of grossly deleted nef genes in HIV-1 infected long-term slow progressors treated with Korean red ginseng - Running title: Grossly deleted nef genes in slow progressors -

  • Cho, Young-K.;Lim, Ji-Y.;Jung, You-S.;Oh, Sun-K.;Lee, Hee-J.;Sung, Heung-Sup
    • Proceedings of the Ginseng society Conference
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    • 2006.05a
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    • pp.40-51
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    • 2006
  • To investigate the association between Korean red ginseng (KRG) intake in HIV-1 infected patients and occurrence of grossly deleted nef genes ($g{\Delta}nef$), we characterized nef genes in 10 long-term slow progressors (LTSP) infected with HIV-1 subtype B and 34 control patients. LTSP was defined whose the annual decrease in CD4 T cells was less than $20/{\mu}l$ over 10 years in the absence of antiretroviral therapy. They were treated with KRG for a prolonged period. Nef genes were amplified from peripheral blood mononuclear cells (PBMC) using nested PCR and products were sequenced directly. Patient CD4 T cell counts decreased from $444{\pm}207/{\mu}l$ to $294{\pm}177/{\mu}l$ over $136{\pm}23$ months of KRG intake. This corresponds to an annual decrease in the level of CD4 T cells of $13.3/{\mu}l$. A total of 479 nef genes were amplified from 137 PBMC samples. Nine out of the 10 patients, 47 (34.3%) out of the 137 samples, and 92 out of the 479 genes revealed $g{\Delta}nef$. The deletion extended outside the nef gene in 25 $g{\Delta}nef$ obtained from 6 patients. The proportion of samples with $g{\Delta}nef$ (34.3%) was significantly higher than 4.8% in control patients (P<0.001). In addition, it significantly increased as the duration of KRG intake prolongs (P<0.01). These data suggest the possibility that occurrence of $g{\Delta}nef$ might be associated with long-term intake of KRG.

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Design and Implementation of Object Reusing Methods for Mobile Vector Map Services (모바일 벡터 지도 서비스를 위한 객체 재사용 기법의 설계 및 구현)

  • Kim, Jin-Deog;Choi, Jin-Oh
    • The KIPS Transactions:PartD
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    • v.10D no.3
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    • pp.359-366
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    • 2003
  • Although the reuse of the cached data for scrolling the map reduces the amount of passed data between client and server, it needs the conversions of data coordinates, selective deletion of objects, cache compaction and object structuring step in the clients. The conversion is a time- intensive operation due to limited resources of mobile phones such as low computing power, small memory. Therefore, in order to control the map efficiently in the vector map service based mobile phones, it is necessary to study the methods which reuse cached objects for reducing wireless network bandwidth and overwhelming the limited resources of mobile phones as well. This paper proposes the methods of reusing pre-received spatial objects for map control in the mobile vector map service system based on client-server architecture. The experiments conducted on the Web GIS systems with real data show that the proposed method is appropriate to map services for mobile phone. We also analyze the advantages and drawbacks between the reuse of cached data and transmission of raw data respectively.

Towards Methionine Overproduction in Corynebacterium glutamicum - Methanethiol and Dimethyldisulfide as Reduced Sulfur Sources

  • Bolten, Christoph J.;Schroder, Hartwig;Dickschat, Jeroen;Wittmann, Christoph
    • Journal of Microbiology and Biotechnology
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    • v.20 no.8
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    • pp.1196-1203
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    • 2010
  • In the present work, methanethiol and dimethyldisulfide were investigated as sulfur sources for methionine synthesis in Corynebacterium glutamicum. In silico pathway analysis predicted a high methionine yield for these reduced compounds, provided that they could be utilized. Wild-type cells were able to grow on both methanethiol and dimethyldisulfide as sole sulfur sources. Isotope labeling studies with mutant strains, exhibiting targeted modification of methionine biosynthesis, gave detailed insight into the underlying pathways involved in the assimilation of methanethiol and dimethyldisulfide. Both sulfur compounds are incorporated as an entire molecule, adding the terminal S-$CH_3$ group to O-acetylhomoserine. In this reaction, methionine is directly formed. MetY (O-acetylhomoserine sulfhydrylase) was identified as the enzyme catalyzing the reaction. The deletion of metY resulted in methionine auxotrophic strains grown on methanethiol or dimethyldisulfide as sole sulfur sources. Plasmid-based overexpression of metY in the ${\Delta}$metY background restored the capacity to grow on methanethiol or dimethyldisulfide as sole sulfur sources. In vitro studies with the C. glutamicum wild type revealed a relatively low activity of MetY for methanethiol (63 mU/mg) and dimethyldisulfide (61 mU/mg). Overexpression of metY increased the in vitro activity to 1,780 mU/mg and was beneficial for methionine production, since the intracellular methionine pool was increased 2-fold in the engineered strain. This positive effect was limited by a depletion of the metY substrate O-acetylhomoserine, suggesting a need for further metabolic engineering targets towards competitive production strains.

Participation of SRE4, an URE1 Enhancer Core Sequence, in the Sterol-Mediated Transcriptional Upregulation of the Human Apolipoprotein E Gene

  • Min, Jung-Hwa;Paik, Young-Ki
    • BMB Reports
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    • v.31 no.6
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    • pp.565-571
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    • 1998
  • The expression of the endogenous human apolipoprotein(apo)E gene was significantly induced when HepG2 cells were treated with exogenous 25-hydroxy-cholesterol. This sterol-mediated apoE gene upregulation appears to require the participation of a positive element for the apoE gene transcription (PET) ( -169/ -140), a core sequence of upstream regulatory element (URE)1 enhancer of the human apoE gene. This PET was renamed as sterol regulatory element (SRE)4 based on its new role as a sensor for the level of intracellular sterol. Furthermore, a gel mobility shift analysis showed that binding activity of the SRE4 binding protein (BP) obtained from HepG2 cells was induced by sterol treatment, while that from either MCF7 or BT20 cells remained unchanged. Binding activity of SRE4BP was also induced in mouse macrophage cells, J774A.1, by sterol treatment, but it was drastically reduced when cells were subjected to treatment of AY-9944, a potent inhibitor for sterol synthesis. However, binding activity of Spl, which is a co-binding protein to the SRE4 region, remained the same in either condition, suggesting that SRE4BP (formally known as PETBP) may be mainly responsible for the sterol-mediated regulation of the apoE gene expression. Deletion analysis of the core binding site of SRE4BP by gel mobility shift assays showed that the minimal sequence of the SRE4BP binding appears to reside between -157 and -140, confirming the identity of SRE4 with the previously determined core sequence of URE1.

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Impacts of Dietary Vitamins and Trace Minerals on Growth and Pork Quality in Finishing Pigs

  • Choi, S.C.;Chae, B.J.;Han, In K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.10
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    • pp.1444-1449
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    • 2001
  • Two feeding trials were conducted to determine the effect of inclusion levels or deletion of vitamin-trace mineral (VM) premixes on growth performance and pork quality in finishing pigs. In exp. 1, a total of ninety-six crossbred pigs $(Landrace\;{\times}\;Yorkshire\;{\times}\;Duroc,\;85.09{\pm}3.12kg)$ were used for a 4-week feeding triaL Treatments were premix supplementation at the level of 50% (Control), 100%, 150%, and 200% of NRC (1998) requirements for vitamins and trace minerals. In exp. 2, a total of one hundred and eight crossbred pigs $(Landrace\;{\times}\;Yorkshire\;{\times}\;Duroc,\;84.76{\pm}0.58kg)$ were used for a 4-week feeding trial. Treatments were premix supplementation at the level of 0% (Control), 200% VM, and 200% vitamin E and Se listed in NRC (1998) requirements. Average daily gain (ADG) and feed/gain (F/G) were the highest at 150% VM addition level (quadratic, p<0.05) among treatments. Dressing percentage and backfat thickness in pigs were not affected by different addition levels of VM premixes. Pork stability in terms of TBARS was linearly (p<0.05) improved as dietary VM premix was increased (exp. 1). ADG, F/G and pork stability (TBARS) were also reduced (p<0.05) when VM premixes were deleted. However, supplementation of vitamin E and Se improved (p<0.05) ADG and pork stability when pigs were fed diets without VM premixes (exp. 2). In conclusion, deleting dietary VM premixes gave negative effects on growth performance and pork quality for the last 4 weeks of finishing period.

Inhibition of Seed Germination and Induction of Systemic Disease Resistance by Pseudomonas chlororaphis O6 Requires Phenazine Production Regulated by the Global Regulator, GacS

  • Kang, Beom-Ryong;Han, Song-Hee;Zdor, Rob E.;Anderson, Anne J.;Spencer, Matt;Yang, Kwang-Yeol;Kim, Yong-Hwan;Lee, Myung-Chul;Cho, Baik-Ho;Kim, Young-Cheol
    • Journal of Microbiology and Biotechnology
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    • v.17 no.4
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    • pp.586-593
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    • 2007
  • Seed coating by a phenazine-producing bacterium, Pseudomonas chlororaphis O6, induced dose-dependent inhibition of germination in wheat and barley seeds, but did not inhibit germination of rice or cucumber seeds. In wheat seedlings grown from inoculated seeds, phenazine production levels near the seed were higher than in the roots. Deletion of the gacS gene reduced transcription from the genes required for phenazine synthesis, the regulatory phzI gene and the biosynthetic phzA gene. The inhibition of seed germination and the induction of systemic disease resistance against a bacterial soft-rot pathogen, Erwinia carotovora subsp. carotovora, were impaired in the gacS and phzA mutants of P chlororaphis O6. Culture filtrates of the gacS and phzA mutants of P. chlororaphis O6 did not inhibit seed germination of wheat, whereas that of the wild-type was inhibitory. Our results showed that the production of phenazines by P. chlororaphis O6 was correlated with reduced germination of barley and wheat seeds, and the level of systemic resistance in tobacco against E. carotovora.