• Title/Summary/Keyword: Flanking region

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Characterization of Korean Cattle Keratin IV Gene

  • Kim, D.Y.;Yu, S.L.;Sang, B.C.;Yu, D.Y.
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.7
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    • pp.1055-1059
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    • 2003
  • Keratins, the constituents of epithelial intermediate filaments, are precisely regulated in a tissue and development specific manner. There are two types of keratin in bovine. The type I is acidic keratin and the type II is neutral/basic keratin. 1.5 kb of 5' flanking sequence of Korean cattle Keratin IV gene, type II keratin (59 kDa), was cloned and sequenced. A symmetrical motif AApuCCAAA are located in a defined region upstream of the TATA box. Proximal SP1, AP1, E-box and CACC elements as the major determinants of transcription are identified. When it was compared to the bovine sequence from -600 bp to ATG upstream, the homology was 97% in nucleotide sequence. Several A and T sequences, located in the promoter region, are deleted in the Korean cattle. An expression vector consisted of Korean cattle Keratin IV gene promoter/SV40 large T antigen was transfected to HaCaT cell (Epithelial keratinocyte). The transformed HaCaT cells showed active proliferation when treated with PDGF (Platelet-derived growth factor) in 0.3% soft agar compared to control cells. These results indicate that Korean cattle Keratin IVgene promoter can be used as a promoter for transfection into epithelial cell.

Transcriptional Induction of a Carbon Starvation Gene during Other Starvation and Stress Challenges in Pseudomonas putida MK1: A Role of a Carbon Starvation Gene in General Starvation and Stress Responses

  • Chitra, Subramanian;Lee, Ho-Sa;Kim, Youngjun
    • Journal of Microbiology
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    • v.37 no.3
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    • pp.141-147
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    • 1999
  • Thirteen transcriptionally-fused carbon starvation mutants, derived from Pseudomonas putida ATCC 12633, were analyzed for their survivability and transcriptional induction profiles upon carbon starvation. One of these mutants, MK114, which exhibited the lowest survivability and the highest induction rate, was selected and further examined under different starvation (nitrogen and phosphate) and stress (osmolarity, H2O2, salts, alcohol, and heat) conditions. Under all tested conditions MK114 induced ${\beta}$-galactosidase activity, implying that the interrupted gene (cst114) is a general starvation and stress response gene. The rate of induction ranged from 2.6-fold for phosphate starvation to 3.7-fold for osmotic shock. The mini-Tn5 flanking DNA was cloned from the chromosome of MK114. The cloned DNA fragment exhibited carbon starvation activity, indicating that this fragment contains a carbon starvation-related promoter region. This region was partially sequenced. Possible physiological roles of Cst114 in a carbon sensing mechanism and in other stress responses are also discussed.

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Molecular characterization of glutathione peroxidase gene from the liver of silver carp, bighead carp and grass carp

  • Li, Guang-Zhao;Liang, Xu-Fang;Yao, Wei;Liao, Wan-Qin;Zhu, Wei-Feng
    • BMB Reports
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    • v.41 no.3
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    • pp.204-209
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    • 2008
  • The cDNAs encoding glutathione peroxidase (GPx) were cloned and sequenced from the liver of three Chinese carps with different tolerance to hepatotoxic microcystins, phyto-planktivorous silver carp (Hypophthalmichthys molitrix) and bighead carp (Aristichthys nobilis), and herbivorous grass carp (Ctenopharyngodon idellus). Using genome walker method, a 750 bp 5'-flanking region of the silver carp GPx gene was obtained, and several potential regulatory elements were identified in the promoter region of the GPx gene. The silver carp GPx gene was widely expressed in all tissues examined. Despite phylogenetic analysis, assigning this newly described carp GPx to the group of mammalian GPx2, the carp GPx seems more similar to GPx1 from a physiological point of view. The constitutive expression pattern of the three carp liver GPx gene, shows a positive relationship with their tolerance to microcystins.

Mapping of Quantitative Trait Loci Affecting Growth Traits in a Japanese Native Chicken Cross

  • Rikimaru, K.;Sasaki, O.;Koizumi, N.;Komatsu, M.;Suzuki, K.;Takahashi, Hideaki
    • Asian-Australasian Journal of Animal Sciences
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    • v.24 no.10
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    • pp.1329-1334
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    • 2011
  • The Hinai-dori is a breed of chicken native to Akita Prefecture, Japan. An $F_2$ resource population produced by crossing low- and high-growth lines of the Hinai-dori breed was analyzed to detect quantitative trait loci (QTL) for growth traits. Highly significant QTLs for body weight at 10 and 14 weeks of age and average daily gain between 4 and 10 weeks and between 10 and 14 weeks of age were accordingly mapped in a common region between ADL0198 and ABR0287 on chromosome 1 and between MCW0240 and ABR0622 on chromosome 4, respectively. A significant QTL for body weight at 4 weeks of age and a significant QTL for average daily gain between 0 and 4 weeks of age were mapped for the first time to the same region flanking ABR0204 and ABR0284 on chromosome 1. These QTLs are good candidates for application in the development of marker-assisted selection strategies for increasing growth efficiencies in the Hinai-dori breed and native breeds of chickens in Asia.

MOLECULAR CLONING OF CHICKEN INTERFERON-GAMMA (닭 인터페론 유전자의 클로닝에 관한 연구)

  • ;Hyun Lillehoj
    • Proceedings of the Korea Society of Poultry Science Conference
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    • 1999.11a
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    • pp.34-50
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    • 1999
  • A cDNA encoding chicken interferon-gamma (chIFN-${\gamma}$) was amplified from P34, a CD4$^{+}$ T-cell hybridoma by reverse transcription-polymerase chain reaction (RT-PCR) and cloned into pUC18. THe sequences of cloned PCR products were determined to confirm the correct cloning. Using this cDNA as probe, chicken genomic library from White Leghorn spleen was screened. Phage clones harboring chicken interferon-gamma (chIFN-${\gamma}$) were isolated and their genomic structure elucidated. The chIFN-${\gamma}$ contains 4 exons and 3 introns spanning over 14 kb, and follows the GT/AG rule for correct splicing at the exon/intron boundaries. The four exons encode 41, 26, 57 and 40 amino acids, respectively, suggesting that the overall structure of IFN-${\gamma}$ is evolutionairly conserved in mammalian and avian species. The 5’-untranslated region and signal sequences are located in exon 1. Several AT-rich sequences located in the fourth exon may indicate a role in mRNA turnover. The 5’-flanking region contains sequences homologous to the potential binding sites for the mammalian transcription factors, activator protein-1(AP-1) activator protein-2(AP-2) cAMP-response element binding protein(CREB), activating transcription factor(ATF), GATA-binding fator(GATA), upstream stimulating factor(USF), This suggests that the mechanisms underlying transcriptional regulation of chicken and mammalian IFN-${\gamma}$ genes may be similar.r.

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Cloning and Characterization of the Urease Gene Cluster of Streptococcus vestibularis ATCC49124

  • Kim Geun-Young;Lee Mann-Hyung
    • Journal of Microbiology and Biotechnology
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    • v.16 no.2
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    • pp.286-290
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    • 2006
  • A genomic library of Streptococcus vestibularis ATCC49124 was constructed in an E. coli plasmid vector, and the urease-positive transformants harboring the urease gene cluster were isolated on Christensen-urea agar plates. The minimal DNA region required for urease activity was located in a 5.6 kb DNA fragment, and a DNA sequence analysis revealed the presence of a partial ureI gene and seven complete open reading frames, corresponding to ureA, B, C, E, F, G, and D, respectively. The nucleotide sequence over the entire ure gene cluster and 3'-end flanking region of S. vestibularis was up to 95% identical to that of S. salivarius, another closely related oral bacterium, and S. thermophilus, isolated from dairy products. The predicted amino acid sequences for the structural peptides were 98-100% identical to the corresponding peptides in S. salivarius and S. thermophilus, respectively, whereas those for the accessory proteins were 96-100% identical. The recombinant E. coli strain containing the S. vestibularis ure gene cluster expressed a high level of the functional urease holoenzyme when grown in a medium supplemented with 1 mM nickel chloride. The enzyme was purified over 49-fold by using DEAE-Sepharose FF, Superdex HR 200, and Mono-Q HR 5/5 column chromatography. The specific activity of the purified enzyme was 2,019 U/mg, and the Michaelis constant ($K_{m}$) of the enzyme was estimated to be 1.4 mM urea. A Superose 6HR gel filtration chromatography study demonstrated that the native molecular weight was about 196 kDa.

Lactosylceramide α2,3-Sialyltransferase Is Induced Via a PKC/ERK/CREB-dependent Pathway in K562 Human Leukemia Cells

  • Choi, Hee-Jung;Park, Young-Guk;Kim, Cheorl-Ho
    • Molecules and Cells
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    • v.23 no.2
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    • pp.138-144
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    • 2007
  • Previously we showed that the human GM3 synthase gene was expressed during the induction of megakaryocytic differentiation in human leukemia K562 cells by phorbol 12-myristate 13-acetate (PMA). In this study we found that treatment of PMA-induced K562 cells with $G{\ddot{o}}6976$, a specific inhibitor of PKC, and U0126, an inhibitor of the extracellular signal-regulated kinase (ERK) reduced expression of GM3 synthase, whereas wortmannin, an inhibitor of phosphoinositide 3-kinase (PI3K) did not. Moreover, activation of ERK and cAMP response element binding protein (CREB) was prevented by pretreatment with $G{\ddot{o}}6976$ and U0126. PMA stimulated the promoter activity of the 5'-flanking region from -177 to -83 region of the GM3 synthase gene, and mutation or deletion of a CREB site located around -143 of the promoter reduced PMA-stimulated promoter activity, as did the inhibitors $G{\ddot{o}}6976$ and U0126. Our results demonstrate that induction of GM3 synthase during megakaryocytic differentiation in PMA-stimulated human leukemia K562 cells depends upon the PKC/ERK/CREB pathway.

Transduction of the Wild-type polA Gene of Escherichia coli K-12 in a ColE1-Derived Mini-Mu Plasmid

  • Parduez, Nagy-Gyorgy;Choi, Yong-Keel;Chung, Young-Sup
    • Korean Journal of Microbiology
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    • v.30 no.2
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    • pp.134-140
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    • 1992
  • Teh $polA^{+}$ gene can be transducted in a multicopy mini-Mu plasmid, but not cloned because the product of this gene is lethal when overproduced. Although, we obtained one surviving cell, in which the ColEl-derived mini-Mu plasmid suffered a spontaneous deletion exactly at the region where the $polA^{+}$ gene was cloned. The $PolA^{+}$ unstream flanking sequence containing the promoter and pribnow-box was delected in vivo ; consequently this gene is not able to be expressed.

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Multiple Age-Associated Mitochondrial DNA Deletions in Mouse Brain

  • Kim, Jin-Sun;Kim, Min-Jung;Kwon, In-Sook;Song, Eun-Sook
    • BMB Reports
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    • v.30 no.1
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    • pp.33-36
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    • 1997
  • Age-dependent deletion of mitochondrial DNA (mtDNA) was detected in mouse brain using PCR method. The size of the deleted fragment was 0.5 kb, 0.9 kb. 1.7 kb and 4.3 kb in the region between cytochrome b gene and ATPase 6 gene. The deleted fragment was increased gradually from 3-month to 22month Direct repeat sequence flanking the deletion in 0.5 kb PCR product was TAAT.

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