• Title/Summary/Keyword: Phosphoglucose isomerase

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Cloning and characterization of phosphoglucose isomerase from Sphingomonas chungbukensis DJ77

  • Tran, Sinh Thi;Le, Dung Tien;Kim, Young-Chang;Shin, Malshik;Choi, Jung-Do
    • BMB Reports
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    • v.42 no.3
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    • pp.172-177
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    • 2009
  • Phosphoglucose isomerase (PGI) is involved in synthesizing extracellular polysaccharide (EPS). The gene encoding PGI in Sphingomonas chungbukensis DJ77 was cloned and expressed in E. coli, and the protein was characterized. The pgi gene from DJ77 is 1,503 nucleotides long with 62% GC content and the deduced amino acid sequence shows strong homology with PGIs from other sources. The molecular masses of PGI subunit and native form were estimated to be 50 kDa and 97 kDa, respectively. Four potentially important residues (H361, R245, E330 and K472) were identified by homology modeling. The mutations, H361A, R245A, E330A, R245K and E330D resulted in decrease in Vmax by hundreds fold, however no significant change in Km was observed. These data suggest that the three residues (H361, R245Aand E330) are likely located in the active site and the size as well as the spatial position of side chains of R245 and E330 are crucial for catalysis.

Studies on the Identification of Turfgrass by Electrophoresis (SDS-PAGE, PAGIF) (전기영동법 (SDS-PAGE, PAGIF)에 의한 잔디 분류에 관한 연구)

  • 박재복;김영후;이수영
    • Asian Journal of Turfgrass Science
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    • v.5 no.1
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    • pp.11-22
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    • 1991
  • This experiment was executed to investigate the possibility of the application of taxonomic method through the isoelectric focusing with polyacrylamide gel and sodium dodecyl sulfate-polyacrylamide gel electrophoresis with seeds in the identification of turfgrasses. The sodium dodecyl sulfate-polyacrylamide gel electrophoresis was used to investigate the pattern of seed proteins which were extracted from 18 cultivars of cool season turfgrass and 4 cultivars of warm season turfgrass. The isoelectric focusing with polyacrylarnide gel was used to investigate the activity of the three isozymes of esterase, peroxidase and phosphoglucose isomerase which were extracted from 18 cultivars of cool season turfgrass and 4 cultivars of warm season turfgrass. The results were summarized as follows. 1. The difference of the patterns of seed proteins was observed with sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The identification of intra-genus was easily detected. 2. The three isozymes of esterase, peroxidase and phosphoglucose isomerase were investigated through isoelectric focusing with polyacrylamide gel. As a result, esterase was most effective among three isozymes in the identification of turfgrass cultivars 3. In the past cultivar identification was primarily based on visual morphological characters, but there was a lot of difficulty. If we should use electrophoresis, we will be able to identifvturfgrass cultivars more effectively.

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Down-regulation of Phosphoglucose Isomerase/Autocrine Motility Factor Enhances Gensenoside Rh2 Pharmacological Action on Leukemia KG1α Cells

  • You, Zhi-Mei;Zhao, Liang;Xia, Jing;Wei, Qiang;Liu, Yu-Min;Liu, Xiao-Yan;Chen, Di-Long;Li, Jing
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.3
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    • pp.1099-1104
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    • 2014
  • Aims and Background: Ginsenoside Rh2, which exerts the potent anticancer action both in vitro and in vivo, is one of the most well characterized ginsenosides extracted from ginseng. Although its effects on cancer are significant, the underlying mechanisms remain unknown. In this study, we sought to elucidate possible links between ginsenoside Rh2 and phosphoglucose isomerase/autocrine motility factor (PGI/AMF). Methods: $KG1{\alpha}$, a leukemia cell line highly expressing PGI/AMF was assessed by western blot analysis and reverse transcription- PCR (RT-PCR) assay after transfection of a small interfering (si)-RNA to silence PGI/AMF. The effect of PGI/AMF on proliferation was measured by typan blue assay and antibody array. A cell counting kit (CCK)-8 and flow cytometry (FCM) were adopted to investigate the effects of Rh2 on PGI/AMF. The relationships between PGI/AMF and Rh2 associated with Akt, mTOR, Raptor, Rag were detected by western blot analysis. Results: KG1${\alpha}$ cells expressed PGI/AMF and its down-regulation significantly inhibited proliferation. The antibody array indicated that the probable mechanism was reduced expression of PARP, State1, SAPK/JNK and Erk1/2, while those of PRAS40 and p38 were up-regulated. Silencing of PGI/AMF enhanced the sensibility of $KG1{\alpha}$ to Rh2 by suppressing the expression of mTOR, Raptor and Akt. Conclusion: These results suggested that ginsenoside Rh2 suppressed the proliferation of $KG1{\alpha}$, the same as down-regulation of PGI/AMF. Down-regulation of PGI/AMF enhanced the pharmacological effects of ginsenoside Rh2 on KG1${\alpha}$ by reducing Akt/mTOR signaling.

Studies on Agronomic and Biochemical Characteristics in Korean Native Rice Cultivars I. Isozyme Variation by Isoelectric Focusing - EST, ME, PGI, HK (한국 벼 재래종의 작물학적 및 생화학적 특성연구 I. 등전점 전기영동법에 의한 Esterase, Malic enzyme, Phosphoglucose isomerase 및 Hexokinase 동위효소 형태 특성)

  • Eun M. Y.;Kim Y. K.;Cho Y. G.;Kim Y. W.;Chung T. Y.;Choi H. C.
    • Proceedings of the Korean Society of Crop Science Conference
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    • 1989.05a
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    • pp.18-19
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    • 1989
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Studies on the Origin of Prunus yedoensis Matsumura 1. A comparative Electrophoretic Study on Wild P. subhirtella in Mt. Hanla, Cultivated P. yedoensis and P. donarium (왕벚의 기원에 관한 연구 1. 한라산자생올벚, 재배왕벚 및 산벚의 전기영동형 비교)

  • 한창열
    • Journal of Plant Biology
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    • v.20 no.1
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    • pp.1-5
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    • 1977
  • Horizontal starch gel electrophoresis in three buffers was used to compare the electrophoretic patterns in three cherry species, wild Prunus subhirtella, cultivated P. yedoensis and P. donarium. Electrophoretic patterns of glutamate oxaloacetate, transaminase-2(GOT-2), malate dehydrogenase-2(MDH-2), and phosphoglucose isomerase(PGI) in three species showed strong evidence that P. yedoensis might be originated by hybridization between P. subhirtella and P. donarium.

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Varietal Classification of Barley by Isozymes and Restriction Fragment Length Polymorphisms (RFLPs) (동위효소 분석과 제한효소 단편 다형화현상을 이용한 보리 품종의 분류)

  • Jin, Byung-Soon;Park, Ro-Dong;Eun, Moo-Young;Lee, Eun-Seop
    • Applied Biological Chemistry
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    • v.36 no.3
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    • pp.139-145
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    • 1993
  • The methods of isoelectric focusing of 4 isozymes in polyacrylamide horizontal slab gels and restriction fragment length polymorphisms (RFLPs) were applied to characterize the biochemical phenotypes of 19 cultivars of barley. Among 19 barley cultivars screened, 7 esterase, 3 phosphoglucose isomerase, 4 peroxidase and 2 alcohol dehydrogenase isozyme phenotypes were distinguished by isoelectric focusing. When purified DNA of each cultivar was digested with restriction enzyme EcoRV and analyzed its RFLPs with barley DNA markers pMSU 51 or pMSU 71, two distinct RFLP patterns were shown. Based on the four isozymes and two RELP polymorphisms, 19 cultivars of barley were classified into 13 biochemical phenotypes. Phylogenetical relationships among 13 biochemical phenotypes classified were determined using Nei's F-statistics and the phylogenetic dendrogram was constructed.

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Isozyme electrophoresis patterns of the liver fluke, Clonorchis sinensis from Kimhae, Korea and from Shenyang, China

  • Park, Gab-Man;Yong, Tai-Woon;Im, Kyung-Il;Lee, Kyu-Je
    • Parasites, Hosts and Diseases
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    • v.38 no.1
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    • pp.45-48
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    • 2000
  • An enzyme analysis of the liver fluke, Clonorchis sinensis from Kimhae, Korea and from Shenyang, China was conducted using a horizontal. starch gel electrophoresis in order to elucidate their genetic relationships. A total of eight enzymes was employed from two different kinds of buffer systems. Two loci from each enzyme of aconitase and esterase (${\alpha}-Na{\;}and{\;}{\beta}-Na$) : and only one locus each from six enzymes, gluucose-6-phosphate dehydrogenase (G6PD), ${\alpha}-glycerophosphate$ dehydrogenase (GPD), 3-hydroxybutyrate dehydrogenase (HBDH), malate dehydrogenase (MDH), phosphoglucose isomerase (PGI), and phosphoglucomutase (PGM) were detected. Most of loci in two populations of C. sinensis showed homozygous monomorphic banding patterns and one of them, GPD was specific as genetic markers between two different populations. However, esterase (${\alpha}-Na$), GPD, HBDH and PGI loci showed polymorphic banding patterns. Two populations of C. sinensis were more closely clustered within the range of genetic identity value of 0.998-1.0. In summarizing the above results, two populations of C. sinensis employed in this study showed mostly monomorphic enzyme protein banding patterns, and genetic differences specific between two populations.

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Identification of differentially expressed genes in the developmental stages from olive flounder Paralichthys olivaceus using an annealing control primer system

  • Kim, Young-Ok;Park, Eun-Mi;Nam, Bo-Hye;Kong, Hee-Jeong;Kim, Woo-Jin;Noh, Jae-Koo;Lee, Sang-Jun;Kim, Kyung-Kil
    • Animal cells and systems
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    • v.14 no.1
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    • pp.25-30
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    • 2010
  • We employed a new and improved differential display reverse transcription-polymerase chain reaction (DDRT-PCR) method, which involves annealing control primers (ACPs), to identify the genes that are specifically or prominently expressed in olive flounder (Paralichthys olivaceus) juveniles (35 days post-hatch; dph) compared to larval-stage (dph 21) flounder. Using 60 ACPs, we identified eight differentially expressed genes (DEGs) and basic local alignment search tool (BLAST) searches revealed eight known genes. Gene expression levels were confirmed by RT-PCR. Phosphoglucose isomerase (PGI) was highly expressed at 21 dph, while nephrosin, myosin light chain (MLC), myosin heavy chain (MHC), carboxypeptidase A, chymotrypsin B, fish-egg protein, and matrix protein were expressed at 35 dph. PGI, MLC, and MHC expression was further analyzed by RT-PCR. The differentially expressed genes identified in this study may provide insights into the molecular basis of development in olive flounder.

Isozyme Analysis on the Allotriploid between Rainbow Trout (Oncorhynchus mykiss) and Coho Salmon (O. kisutch) (무지개송어와 은연어간 잡종3배체의 부화자어에 대한 동위효소 분석)

  • HONG Kyung Pyo;MYOUNG Jung-Goo;KIM Pyong Kih;SON Jin-Ki
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.29 no.2
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    • pp.256-261
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    • 1996
  • For the purpose of identification of inheritance in allotriploid between rainbow trout (Oncorhynchus mykiss) and coho salmon (O. kisutch), five isozymes, lactate dehydrogenase (LDH), malate dehydrogenase (MDH), isocitrate dehydrogenase (IDH), phosphoglucose isomerase (PGI) and phosphoglucomutase (PGM) from skeletal muscle in two species and their allotriploid were analyzed. All of these loci showed differences between two species and their allotriploid except PGI. Generally, coho salmon was more monomorphic in these isozyme loci than rainbow trout. Their allotriploids showed intermediate patterns between the parental species in those isozyme loci except PGI. As a result of this study, LDH, MDH, IDH and PSM may be used as useful genetic markers in these two species, and they also be of use in studying hybrid and allotriploid in salmonids.

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A Biochemical Study for the Development of Genetic Marker on Salmonids in Korea (한국산 연어류에서 Genetic Marker 개발을 위한 생화학적 연구)

  • HONG Kyung-Pyo;MYOUNG Jung-Goo;SON Jin-Ki;PARK Chul-Won
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.27 no.1
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    • pp.83-88
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    • 1994
  • For the purpose of genetic stock indentification of three species of salmonid fishs and their hybrid, lactate dehydrogenase(LDH), malate dehydrogenase(MDH), isocitrate dehydrogenase(IDH), a-gylycerophosphate dehydrogenase(a-GPDH), malic enzyme(ME), 6-phospho-gluconate dehydrogenase(6-PGD), phosphoglucose isomerase(PGI) and phospho-glucomutase(PGM) from skeletal muscle, liver, heart and gill tissues in all three species were analyzed. Chum and masu salmon showed no polymorphic patterns in all isozyme loci, however rainbow trout were found to have polymorphic patterns at MDH-B, LDH and IDH loci. Especially, significant differences were found at MDH-B loci between the three species and the IDH patterns of rainbow trout were also different from the other two species. These loci therefore can be utilized as efficient genetic markers for the identification of hybrids and improve the efficiency of fish breeding. There was no difference except PGI between diploid and triploid isozyme patterns but PGI showed some potential as a marker for triploid in masu salmon.

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