• Title/Summary/Keyword: GAPDH

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Selection of suitable reference gene for gene expression studies of porcine ovaries under different conditions in quantitative reverse transcription polymerase chain reaction assay

  • Kim, Hwan-Deuk;Jeon, Hye-Jin;Jang, Min;Bae, Seul-Gi;Yun, Sung-Ho;Han, Jee-Eun;Kim, Seung-Joon;Lee, Won-Jae
    • Journal of Animal Reproduction and Biotechnology
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    • v.37 no.2
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    • pp.96-105
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    • 2022
  • The ovary undergoes substantial physiological changes along with estrus phase to mediate negative/positive feedback to the upstream reproductive tissues and to play a role in producing a fertilizable oocyte in the developing follicles. However, the disorder of estrus cycle in female can lead to diseases, such as cystic ovary which is directly associated with decline of overall reproductive performance. In gene expression studies of ovaries, quantitative reverse transcription polymerase chain reaction (qPCR) assay has been widely applied. During this assay, although normalization of target genes against reference genes (RGs) has been indispensably conducted, the expression of RGs is also variable in each experimental condition which can result in false conclusion. Because the understanding for stable RG in porcine ovaries was still limited, we attempted to assess the stability of RGs from the pool of ten commonly used RGs (18S, B2M, PPIA, RPL4, SDHA, ACTB, GAPDH, HPRT1, YWHAZ, and TBP) in the porcine ovaries under different estrus phase (follicular and luteal phase) and cystic condition, using stable RG-finding programs (geNorm, Normfinder, and BestKeeper). The significant (p < 0.01) differences in Ct values of RGs in the porcine ovaries under different conditions were identified. In assessing the stability of RGs, three programs comprehensively agreed that TBP and YWHAZ were suitable RGs to study porcine ovaries under different conditions but ACTB and GAPDH were inappropriate RGs in this experimental condition. We hope that these results contribute to plan the experiment design in the field of reproductive physiology in pigs as reference data.

Identification of Differentially Expressed Genes in the Dicer 1 Knock-down Mouse Embryos using Microarray

  • Lee, Jae-Dal;Cui, Xiang-Shun
    • Reproductive and Developmental Biology
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    • v.32 no.4
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    • pp.229-235
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    • 2008
  • Silencing of Dicer1 by siRNA did not inhibit development up to the blastocyst stage, but decreased expression of selected transcription factors, including Oct-4, Sox2 and Nanog, suggesting that Dicer1 gene expression is associated with differentiation processes at the blastocyst stage (Cui et al., 2007). In order to get insights into genes which may be linked with microRNA system, we compared gene expression profiles in Gapdh and Dicer1 siRNA-microinjected blastocysts using the Applied Biosystem microarray technology. Our data showed that 397 and 737 out of 16354 genes were up- and down-regulated, respectively, following siRNA microinjection (p<0.05), including 24 up- and 28 down-regulated transcription factors. Identification of genes that are preferentially expressed at particular Dicer1 knock down embryos provides insights into the complex gene regulatory networks that drive differentiation processes in embryos at blastocyst stage.

Proteome Analysis of Recombinant CHO Cells Under Hyperosmotic Stress

  • Lee, Mun-Su;Kim, Gyeong-Uk;Kim, Yeong-Hwan;Lee, Gyun-Min
    • 한국생물공학회:학술대회논문집
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    • 2003.04a
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    • pp.311-314
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    • 2003
  • Under hyperosmotic stress, rCHO cells display decreased specific growth rate $({\mu})$ and increased specific antibody productivity $(q_{Ab})$. The effects of hyperosmotic stress on batch culture cellular dynamics are not well understood. To this end, we conducted a proteome profile of rCHO cells, using 2D-gel, MALDI-TOF-MS and MS/MS. As a result, the proteome profile of rCHO cells could be established using 41 identified proteins. Based on this proteome profile of rCHO cells, we have found at least 8 differently expressed spots at hyperosmotic osmolality (450 mOsm/kg). Among these spots, two metabolic enzymes were found to be up-regulated (pyruvate kinase and GAPDH), while down-regulated protein was identified as tubulin. It shows that hyperosmotic stress can alter metabolic state, by up-regulated activities of two glycolysis enzymes, which could lead to activate the generation of metabolic energy. Tubulin expression was down-regulated, suggesting a reduction of cell division. Finally, the increased conversion energy could leads to improve overall productivity.

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Morphology and Molecular Characterization of a Fungus from the Alternaria alternata Species Complex Causing Black Spots on Pyrus sinkiangensis (Koerle pear)

  • Aung, Sein Lai Lai;Liu, Hai Feng;Pei, Dong Fang;Lu, Bing Bin;Oo, May Moe;Deng, Jian Xin
    • Mycobiology
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    • v.48 no.3
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    • pp.233-239
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    • 2020
  • A small-spored Alternaria was found from black spots of storaged Koerle pear (Pyrus sinkiangensis), one of the economically important fruit in Xinjiang province, China. The morphology is similar to A. limoniasperae but obviously different in secondary conidiophores and conidial septa. A phylogenetic analysis using sequence datasets of ITS, GAPDH, TEF1, RPB2, Alt a1, OPA10-2, and EndoPG genes revealed that it belonged to the Alternaria alternata complex group. Pathogenicity tests illustrated that the fungus was the causal pathogen of black spot on Koerle pear fruit.

Archvillin C-Terminus-Binding Proteins in Human Skeletal Muscle

  • Chang, Goo-Rak
    • Biomedical Science Letters
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    • v.16 no.4
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    • pp.207-212
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    • 2010
  • Archivillin, a muscle-specific isoform of supervillin, is a component of the costameric cytoskeleton of muscle cells. The purpose of this study was to determine which protein in the skeletal muscle collaborates with archvillin C-terminus. For this purpose, a yeast two-hybrid screening of human skeletal muscle cDNA library was performed using the C-terminal region of archvillin as bait. This study shows that seven human skeletal muscle proteins, namely, nebulin, xeplin, archvillin, GAPDH, TOX4, PITRM1, and YME1L1 interact with archvillin C-terminus. Especially, xeplin is a newly discovered protein interacts with archvillin C-terminus. These results indicate that archvillin C-terminus acts as a bridge between nebulin and xeplin at costameres. Archvillin C-terminal region interacts with nebulin C-terminal region at Z-discs and interacts with xeplin at the vicinity of sarcolemma. I propose that these interactions may contribute to formation of costameric structure and muscle contraction.

Allozyme Variability in Tree of Genus Semisulcospira(Prosobranchia: Pleuroceridae) (다슬기속 3종(Prosobranchia: Pleuroceridae)에서의 도위효소 변이)

  • 정영헌;박준우;정평림;박갑만;김재진;민득영
    • The Korean Journal of Malacology
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    • v.15 no.1
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    • pp.13-20
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    • 1999
  • A horizontal starch gel electrophoresis for enzyme proteins extracted from three Korean species and one Chinese species of Semisulcospira was carried out in order to elucidate their genetic relationships. A total of 10 enzymes were employed in three different of buffer systems. Two loci from each enzyme of GAPDH, GOT, ICDH, IDH and PEP(VL); three loci from each of three enzymes, EST, PEP(LGG) and PGDH; and five loci from GPI were observed. Most of the loci in three pleurocerid species employed showed homozygous monomorphic banding patterns and some of them were specific as genetic markers between two different species. However, EST-2, PEP(LGG-3) and PGDH-1 loci in Korean S. libertina and PEP(LGG-3), PGM-1 and PGM-2 loci in Chinese S. libertina showed polymorphic banding patterns. Three Korean Semisulcospira species including S. libertina were more closely clustered in a dendrogram within the range of genetic identity values of 0.818-0.936, and these clusters were lineated with Chinese S. libertina at the value of 0.621.

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Morphology and Molecular Characterization of Alternaria argyranthemi on Chrysanthemum coronarium in China

  • Luo, Huan;Xia, Zhen Zhou;Chen, Yun Yun;Zhou, Yi;Deng, Jian Xin
    • Mycobiology
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    • v.46 no.3
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    • pp.278-282
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    • 2018
  • Chrysanthemum coronarium is an economically important plant in Asia, and used medicinally, ornamentally and as a vegetable. In April 2017, leaf spot disease on C. coronarium was observed in Shiyan, Hubei, China. A single-spore isolate was obtained and identified based on morphology and sequence analysis using four regions (rDNA ITS, GAPDH, $EF-1{\alpha}$, and RPB2). The results indicated that the fungus is Alternaria argyranthemi. The pathogenicity tests revealed that the species could cause severe leaf spot and blight disease on the host. This is the first report of leaf spot disease on C. coronarium caused by A. argyranthemi in the world, which is also a new record of Alternaria species in China.

Simultaneous Saccharification of Inulin and Ethanol Fermentation by Recombinant Saccharomyces cerevisiae Secreting Inulinase

  • Kim, Youn-Hee;Nam, Soo-Wan;Chung, Bong-Hyun
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.3 no.2
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    • pp.55-60
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    • 1998
  • Various Saccharomyces cerevisiae strains were transformed with a 2 ${\mu}$-based multicopy expression plasmid, pYIGP, carrying Kluyveromyces marxianus inulinase gene under the control of GAPDH promoter. Among then two strains, SEY2102 and 2805, showed high levels of cell growth and inulinase expression, and were selected to study their fermentation properties on inulin. Jerusalem artichoke inulin was more effective for cell growth (10∼11 g-dry wt./L at 48 hr) and inulinase expression (1.0 units/mL with SEY2102/pYIGP and 2.5 units/mL with 2805/pYIGP) than other inulin sources such as dahlia and chicory. It was also found that maximal ethanol production of 9 g/L was obtained from Jerusalem artichoke inulin at the early stationary phase (around 30 hr), indicating that recombinant S. cerevisiae cells secreting exoinulinase could be used for the simultaneous saccharification of inulin and ethanol fermentation.

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A New Record and Characterization of Asparagus Purple Spot Caused by Stemphylium vesicarium in Korea

  • Han, Joon-Hee;Shin, Jong-Hwan;Fu, Teng;Kim, Kyoung Su
    • Mycobiology
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    • v.47 no.1
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    • pp.120-125
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    • 2019
  • In 2017, small, elliptical, brownish purple spots on spears and ferns of asparagus were found in fields of Gangwon-do. The isolated fungal species was identified as an ascomycete Stemphylium vesicarium based on morphological characteristics and molecular phylogenic analyses including nucleotide sequences of the internal transcribed spacer (ITS), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and cytochrome b (cytb). A pathogenicity test revealed that S. vesicarium was the causal agent of purple spot disease on asparagus. The occurrence of purple spots caused by S. vesicarium on asparagus is the first report in Korea.