• Title/Summary/Keyword: Gal1

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Comparative N-Linked Glycan Analysis of Wild-Type and α1,3-Galactosyltransferase Gene Knock-Out Pig Fibroblasts Using Mass Spectrometry Approaches

  • Park, Hae-Min;Kim, Yoon-Woo;Kim, Kyoung-Jin;Kim, Young June;Yang, Yung-Hun;Jin, Jang Mi;Kim, Young Hwan;Kim, Byung-Gee;Shim, Hosup;Kim, Yun-Gon
    • Molecules and Cells
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    • v.38 no.1
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    • pp.65-74
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    • 2015
  • Carbohydrate antigens expressed on pig cells are considered to be major barriers in pig-to-human xenotransplantation. Even after ${\alpha}1,3$-galactosyltransferase gene knock-out (GalT-KO) pigs are generated, potential non-Gal antigens are still existed. However, to the best of our knowledge there is no extensive study analyzing N-glycans expressed on the GalT-KO pig tissues or cells. Here, we identified and quantified totally 47 N-glycans from wild-type (WT) and GalT-KO pig fibroblasts using mass spectrometry. First, our results confirmed the absence of galactose-alpha-1,3-galactose (${\alpha}$-Gal) residue in the GalT-KO pig cells. Interestingly, we showed that the level of overall fucosylated N-glycans from GalT-KO pig fibroblasts is much higher than from WT pig fibroblasts. Moreover, the relative quantity of the N-glycolylneuraminic acid (NeuGc) antigen is slightly higher in the GalT-KO pigs. Thus, this study will contribute to a better understanding of cellular glycan alterations on GalT-KO pigs for successful xenotransplantation.

STATISTICS OF GRAVITATIONAL LENSING BY A GALAXY IN CLUSTER OR IN FIELD

  • YOON SO-YOON;PARK MYEONG-GU
    • Journal of The Korean Astronomical Society
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    • v.29 no.2
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    • pp.119-136
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    • 1996
  • To examine the effect of neighboring galaxies on the gravitational lensing statistics, we performed numerical simulations of lensing by many galaxies. The models consist of a galaxy in the rich cluster like Coma, or a galaxy surrounded by field galaxies in $\Omega_0 = 1$ universe with $\Omega_{gal} = 0.1,\;\Omega_{gal} = 0.3\;or\;\Omega_{gal}=1.0\;,\;where\;\Omega_{gal}$ is the total mass in galaxies. Field galaxies either have the same mass or follow Schechter luminosity function and luminosity-velocity relation. Each lensing galaxy is assumed to be singular isothermal sphere (SIS) with finite cutoff radius. In most simulations, the lensing is mainly due to the single galaxy. But in $\Omega_{gal} = 3$ universe, one out of five simulations have 'collective lensing' event in which more than two galaxies collectively produce multiple images. These cases cannot be incorporated into the simple 'standard' lensing statistics calculations. In cases where 'collective lensing' does not occur, distribution of image separation changes from delta function to bimodal distribution due to shear induced by the surrounding galaxies. The amount of spread in the distribution is from a few $\%\;up\;to\;50\%$ of the mean image separation in case when the galaxy is in the Coma-like cluster or when the galaxy is in the field with $\Omega_{gal} = 0.1\;or\;\Omega_{gal}=0.3.$ The mean of the image separation changes less than $5\%$ compared with a single lens case. Cross section for multiple image lensing turns out to be relatively insensitive to the presence of the neighboring galaxies, changing less than $5\%$ for Coma-like cluster and $\Omega_{gal}=0.1,\;0.3$ universe cases. So we conclude that Coma-like cluster or field galaxies whose total mass density $\Omega_{gal}<0.3$ do not significantly affect the probability of multiple image lensing if we exclude the 'collective lensing' cases. However, the distribution of the image separations can be significantly affected especially if the 'collective lensing' cases are included. Therefore, the effects of surrounding galaxies may not be negligible when statistics of lensing is used to deduce the cosmological informations.

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Expression of the Galactose Mutarotase Gene from Lactococcus lactis ssp. lactis ATCC7962 in Escherichia coli

  • Lee, Jong-Hoon;Choi, Jae-Yeon;Lee, Jung-Min;Kim, Jeong-Hwan;Chang, Hae-Choon;Chung, Dae-Kyun;Lee, Hyong-Joo
    • Journal of Microbiology and Biotechnology
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    • v.10 no.6
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    • pp.840-843
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    • 2000
  • The structure of gal/lac operon of Lactococcus lactis ssp. lactis ATCC7962 was partially characterized and the gene (galM) encoding galactose mutarotase was cloned together with the order; galA-galM-galK-galT. The galM was found to be 1,027 bp in length and encoded the protein of 37,609 Da calculated molecular mass. The deduced amino acid sequence showed a homology with GalM proteins from several other microorganisms. Thus, the galM gene was expressed in Escherichia coli and the product was identified as a 38 kDa protein which corresponded to the size estimated from DNA sequence. mutarotase activity of the IPTG inducedrecombinant was 2.7 times increased against that of the host strain.

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Overproduction and High Level Secretion of Glucose Oxidase in Saccharomyces cerevisiae (Glucose Oxidase의 Saccharomyces cerevisiae에서의 대량생산 및 고효율 분비)

  • 홍성용;최희경;이영호;백운화;정준기
    • Microbiology and Biotechnology Letters
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    • v.26 no.1
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    • pp.68-75
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    • 1998
  • The overproduction and high level secretion of Glucose Oxidase (GOD) from A. niger in S. cerevisiae was carried out by cloning GOD gene. For this purpose, using two different strong promoters (ADH1 promoter, GAL10 promoter) and signal sequences (${alpha}$-MF signal sequence of S. cerevisiae and ${alpha}$-amylase signal sequence of A. oryzae) and GAL7- and GOD terminator, four expression vectors were constructed. All the expression vectors were transformed in S. cerevisiae 2805 using auxotroph method. By the flask culture, transformants of pGAL expression vector series containing GAL 10 promotor showed much higher GOD productivity than transformants of pADH expression vector series containing ADH1 promoter Transformants of pGALGO2 containing GAL10 promotor and ${alpha}$-amylase signal sequence has shown the best productivity of GOD ($GOD_{total}$: 10.3 unit/mL, $GOD_{ex}$: 8.7 unit/mL) at 115 hr. This value was three fold higher than that of pGALGO1 containing GAL 10 promotor and ${alpha}$-MF signal sequence, even if the same promotor was involved. Through the ${alpha}$-amylase signal sequence of A. oryzae, GOD was secreted much more than the case of ${alpha}$-MF signal sequence from S. cerevisiae. These results suggest that signal sequence may play a important roles in not only the secretion but also the overproduction of foreign protein. Secretion rate of GOD in pGALGO1 and pGALGO2 was 89% and 84%, respectively, Because of the overglycosylation in S. cerevisiae the molecular weight of recombinant GOD in S. cerevisiae was much larger (250 kDa) than that of nature GOD in A. niger (170 kDa).

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The Optimization of Recombinant Protein Production using S. cerevisiae Mutant Y334 Suitable for GAL Promoter (GAL promoter에 적합한 효모변이주 Y334를 이용한 재조합 단백질 생산 최적화 방법 개발)

  • 강환구;전희진;이문원
    • KSBB Journal
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    • v.15 no.2
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    • pp.181-187
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    • 2000
  • The production of heterologous protein using GAL promoter in conventional S. cerevisiae has several problems to s이ve for c commercialization. In this research, S. cerevisiae mutant(reg1-501, gaI1), which cannot use galactose and has alleviated g glucose repression level, is used as host for optimizing induction of GAL promoter. In this experiment, the effects of specific g growth rate on specific recombinant protein expression rate were tested in both cases and optimum fed batch fermentation m method was obtained in both cases. Through these experiments, optimum condition of recombinant protein production by G GAL promoter using S. cerevisiae mutant (reg1-501, gal1) were found.

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젖산생성균의 .betha.-galactosidase의 생화학 및 분자생물학적 특성

  • 민해기
    • The Microorganisms and Industry
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    • v.19 no.1
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    • pp.8-17
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    • 1993
  • 젖산생성균의 .betha.-gal의 생성과 Bif. longum KCTC 3215에 의한 .betha.-gal 생산, 정제 및 특성에 관한 연구와 젖산생성균의 .betha.-gal 유전자의 클로닝 및 대장균에의 발현과 Str.thermophilus SKD 1006의 Lac Z 유전자를 비교 분석하였다.

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Hydrogen Bonds in GlcNAc( β1,3)Gal( β)OMe in DMSO Studied by NMR Spectroscopy and Molecular Dynamics Simulations

  • Shim, Gyu-Chang;Shin, Jae-Min;Kim, Yang-Mee
    • Bulletin of the Korean Chemical Society
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    • v.25 no.2
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    • pp.198-202
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    • 2004
  • Hydrogen bond is an important factor in the structures of carbohydrates. Because of great strength, short range, and strong angular dependence, hydrogen bonding is an important factor stabilizing the structure of carbohydrate. In this study, conformational properties and the hydrogen bonds in GlcNAc( ${\beta}$1,3)Gal(${\beta}$)OMe in DMSO are investigated through NMR spectroscopy and molecular dynamics simulation. Lowest energy structure in the adiabatic energy map was utilized as an initial structure for the molecular dynamics simulations in DMSO. NOEs, temperature coefficients, SIMPLE NMR data, and molecular dynamics simulations proved that there is a strong intramolecular hydrogen bond between O7' and HO3' in GlcNAc( ${\beta}$1,3)Gal(${\beta}$)OMe in DMSO. In aqueous solution, water molecule makes intermolecular hydrogen bonds with the disaccharides and there was no intramolecular hydrogen bonds in water. Since DMSO molecule is too big to be inserted deep into GlcNAc(${\beta}$1,3)Gal(${\beta}$)OMe, DMSO can not make strong intermolecular hydrogen bonding with carbohydrate and increases the ability of O7' in GlcNAc(${\beta}$1,3)Gal(${\beta}$)OMe to participate in intramolecular hydrogen bonding. Molecular dynamics simulation in conjunction with NMR experiments proves to be efficient way to investigate the intramolecular hydrogen bonding existed in carbohydrate.

GALOIS GROUPS OF MODULES AND INVERSE POLYNOMIAL MODULES

  • Park, Sang-Won;Jeong, Jin-Sun
    • Bulletin of the Korean Mathematical Society
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    • v.44 no.2
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    • pp.225-231
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    • 2007
  • Given an injective envelope E of a left R-module M, there is an associative Galois group Gal$({\phi})$. Let R be a left noetherian ring and E be an injective envelope of M, then there is an injective envelope $E[x^{-1}]$ of an inverse polynomial module $M[x^{-1}]$ as a left R[x]-module and we can define an associative Galois group Gal$({\phi}[x^{-1}])$. In this paper we describe the relations between Gal$({\phi})$ and Gal$({\phi}[x^{-1}])$. Then we extend the Galois group of inverse polynomial module and can get Gal$({\phi}[x^{-s}])$, where S is a submonoid of $\mathbb{N}$ (the set of all natural numbers).

Natural and synthetic pathogen associated molecular patterns modulate galectin expression in cow blood

  • Asiamah, Emmanuel Kwaku;Ekwemalor, Kingsley;Adjei-Fremah, Sarah;Osei, Bertha;Newman, Robert;Worku, Mulumebet
    • Journal of Animal Science and Technology
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    • v.61 no.5
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    • pp.245-253
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    • 2019
  • Pathogen-associated Molecular Patterns (PAMPs) are highly conserved structural motifs that are recognized by Pathogen Recognition receptors (PRRs) to initiate immune responses. Infection by these pathogens and the immune response to PAMPS such as lipopolysaccharide (LPS), Peptidoglycan (PGN), bacterial oligodeoxynucleotides [CpG oligodeoxynucleotides 2006 (CpG ODN2006) and CpG oligodeoxynucleotides 2216 (CpG ODN2216)], and viral RNA Polyinosinic-Polycytidylic Acid (Poly I:C), are associated with infectious and metabolic diseases in animals impacting health and production. It is established that PAMPs mediate the production of cytokines by binding to PRRs such as Toll-like receptors (TLR) on immune cells. Galectins (Gal) are carbohydrate-binding proteins that when expressed play essential roles in the resolution of infectious and metabolic diseases. Thus it is important to determine if the expression of galectin gene (LGALS) and Gal secretion in blood are affected by exposure to LPS and PGN, PolyI:C and bacterial CpG ODNs. LPS increased transcription of LGALS4 and 12 (2.5 and 2.02 folds respectively) and decreased secretion of Gal 4 (p < 0.05). PGN increased transcription of LGALS-1, -2, -3, -4, -7, and -12 (3.0, 2.3, 2.0, 4.1, 3.3, and 2.4 folds respectively) and secretion of Gal-8 and Gal-9 (p < 0.05). Poly I:C tended to increase the transcription of LGALS1, LGALS4, and LGALS8 (1.78, 1.88, and 1.73 folds respectively). Secretion of Gal-1, -3, -8 and nine were significantly increased in treated samples compared to control (p < 0.05). CpG ODN2006 did not cause any significant fold changes in LGALS transcription (FC < 2) but increased secretion of Gal-1, and-3 (p < 0.05) in plasma compared to control. Gal-4 was however reduced in plasma (p < 0.05). CpG ODN2216 increased transcription of LGALS1 and LGALS3 (3.8 and 1.6 folds respectively), but reduced LGALS2, LGALS4, LGALS7, and LGALS12 (-1.9, -2.0, -2.0 and; -2.7 folds respectively). Secretion of Gal-2 and -3 in plasma was increased compared to control (p < 0.05). Gal-4 secretion was reduced in plasma (p < 0.05). The results demonstrate that PAMPs differentially modulate galectin transcription and translation of galectins in cow blood.

Hepatoprotective Effects of Oyster Hydrolysate on Lipopolysaccharide/D-Galactosamine-Induced Acute Liver Injury in Mice (Lipopolysaccharide/D-Galactosamine에 의해 유도된 급성 간 손상 생쥐모델에서 굴가수분해물의 간 보호 효과)

  • Ryu, Ji Hyeon;Kim, Eun-Jin;Xie, Chengliang;Nyiramana, Marie Merci;Siregar, Adrian S.;Park, Si-Hyang;Cho, Soo Buem;Song, Dae Hyun;Kim, Nam-Gil;Choi, Yeung Joon;Kang, Sang Soo;Kang, Dawon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.46 no.6
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    • pp.659-670
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    • 2017
  • Oxidative stress and inflammation are key factors responsible for progression of liver injury. A variety of functions of oyster hydrolysate (OH) are affected by their antioxidant and anti-inflammatory activities. However, little is known regarding the effects of OH on a liver injury model. This study was performed to evaluate the effects of OH on acute liver injury induced by lipopolysaccharide/D-galactosamine (LPS/D-GalN) in mice. Experimental groups were divided into six groups as follows (each group, n=10): control (saline), LPS/D-GalN, LPS/D-GalN+OH (100 mg/kg), LPS/D-GalN+OH (200 mg/kg), LPS/D-GalN+OH (400 mg/kg), and LPS/D-GalN+silymarin (25 mg/kg, positive control). The experimental acute liver injury model was induced with LPS ($1{\mu}g/kg$) and D-GalN (400 mg/kg). We first analyzed antioxidant and anti-inflammatory activities in OH. OH showed high DPPH and ABTS radical scavenging activities and reduced ROS generation in Chang cells in a dose-dependent manner. In addition, OH showed anti-inflammatory activities, such as inhibition of cyclooxygenase-2 and 5-lipooxygenase. Treatment with OH down-regulated tumor necrosis factor $(TNF)-{\alpha}$, interleukin (IL)-6, and $IL-1{\alpha}$ expression levels in LPS-stimulated RAW264.7 cells. OH significantly reduced LPS/D-GalN-induced increases in the concentrations of alanine transaminase and aspartate aminotransferase in serum. In the LPS/D-GalN group, liver tissues exhibited apoptosis of hepatocytes with hemorrhages. These pathological alterations were ameliorated by OH treatment. Consistently, hepatic catalase activity was low in the LPS/D-GalN group compared to the control group, and catalase activity was significantly restored by OH treatment (P<0.05). Furthermore, OH markedly reduced the LPS/D-GalN-induced increase in $TNF-{\alpha}$, $IL-1{\beta}$, and IL-6 levels in liver tissue. Taken together, these results show that OH has hepatoprotective effects on LPS/D-GalN-induced acute liver injury via inhibition of oxidative stress and inflammation, suggesting that OH could be used as a health functional food and potential therapeutic agent for acute liver injury.