• Title/Summary/Keyword: Over-expression

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Veri cation of Improving a Clustering Algorith for Microarray Data with Missing Values

  • Kim, Su-Young
    • The Korean Journal of Applied Statistics
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    • v.24 no.2
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    • pp.315-321
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    • 2011
  • Gene expression microarray data often include multiple missing values. Most gene expression analysis (including gene clustering analysis); however, require a complete data matric as an input. In ordinary clustering methods, just a single missing value makes one abandon the whole data of a gene even if the rest of data for that gene was intact. The quality of analysis may decrease seriously as the missing rate is increased. In the opposite aspect, the imputation of missing value may result in an artifact that reduces the reliability of the analysis. To clarify this contradiction in microarray clustering analysis, this paper compared the accuracy of clustering with and without imputation over several microarray data having different missing rates. This paper also tested the clustering efficiency of several imputation methods including our propose algorithm. The results showed it is worthwhile to check the clustering result in this alternative way without any imputed data for the imperfect microarray data.

Secretion of Human Angiogenin into Periplasm and Culture Medium with Its Eukaryotic Signal Sequence by Escherichia coli

  • Jung, Woo-Jung;Choi, Suk-Jung
    • BMB Reports
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    • v.30 no.1
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    • pp.80-84
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    • 1997
  • The synthesis and secretion of human angiogenin in E. coli by the natural leader sequence has been studied. We constructed a recombinant plasmid containing human angiogenin cDNA which encompassed all the coding region including leader sequence required for secretion. The recombinant plasmid was introduced into a suitable E. coli host. The angiogenin was detected in the culture medium and periplasm upon the induction of gene expression. The molecular weight of the secreted angiogenin was identical to that of authentic angiogenin purfied from human plasma when estimated by SDS-PAGE and immunoblotting. showing that the natural leader sequence was recognized and processed by the secretion machinery of E. coli. The angiogenin concentration in the culture medium reached a maximum within 2 h when expressed at $37^{\circ}C$ with 0.02~2 mM IPTG. In contrast, the expression level increased gradually over time up to 11 h at $23^{\circ}C$ with 0.002~2 mM IPTG and at $37^{\circ}C$ with 0.002 mM IPTG.

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Expression of Human Stem Cell Factor with Recombinant Baculovirus in BmN Cell Line and Silkworm

  • Xijie, Guo;Yongfeng, Jin;Mingguan, Yang;Yaozhou, Zhang
    • International Journal of Industrial Entomology and Biomaterials
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    • v.4 no.1
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    • pp.51-56
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    • 2002
  • A recombinant transfer vector pBacSCF was constructed by inserting huamn stem cell factor (hSCF) cDNA into plasmid pBacPAK8. BmN cells were co-transfected with modified Bombyx mori, nuclear polyhedrosis virus (BmBacPAK) DNA and the recmbinant transfer vector to construct a recombinant baculovirus containing hSCE gene. DNA dot blotting and RNA dot blotting demonstrated that the hSCE gene was contained in the recombinant virus and transcribed. The recombinant baculovirus was infectious to BmN cells and to silkworm. SDS-PAGE analysis showed a specific band of expressed product in the extract of infected cells and in the heamolymph of infected larvae. Bioactivity of the recombinant hSCE was determined with W-1 cell line and MTT colorimetric method in synergy with interlukin-3 (IL-3). These results revealed that the hSCF gene was over-expressed in cultured cells and lavae of silkworm.

MDMA (Ecstasy) Induces Egr-1 Expression and Inhibits Neuronal Differentiation

  • Lee, Ji-Hae;Kim, Sung-Tae;Choi, Don-Chan;Lee, Seung-Hoon
    • Development and Reproduction
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    • v.15 no.2
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    • pp.173-178
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    • 2011
  • The amphetamine derivative 3,4-methylenedioxymethamphetamine (MDMA) is a potent monoaminergic neurotoxin with the potential to cause serotonergic neurotoxicity, but has become a popular recreational drug. Little has been known about the cellular effects induced by MDMA. This report shows that MDMA inhibits neuronal cell growth and differentiation. MDMA suppressed neuronal cell growth. The results of quantitative real-time PCR analysis showed that Egr-1 expression is elevated in mouse embryo and neuroblastoma cells after MDMA treatment. Transiently transfected Egr-1 interfered with the neuronal differentiation of neuroblastoma cells such as SH-SY5Y and PC12 cells. These findings provide evidence that the abuse of MDMA during pregnancy may impair neuronal development via an induction of Egr-1 over-expression.

Protein Expression Profiling of Infected Murine Macrophage Cells (RAW 264.7) by Bacillus anthracis Spores

  • Seo Gwi-Moon;Nam Jeong-Ah;Oh Kwang-Gun;Chai Young-Gyu
    • Proceedings of the Microbiological Society of Korea Conference
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    • 2003.05a
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    • pp.77-79
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    • 2003
  • Current therapeutic strategies far anthrax have had no significant impact on anthrax mortality over the last several decades. This study used a matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) discovery platform to generate protein expression profiles in search of overexpressed proteins in murine macrophage cells (RAW264.7) which infected with Bacillus anthracis spores as potentially novel molecular targets. Two differentially expressed proteins were identified in infected murine macrophage cells as Syndapin and CDC46, respectively. Syndapins are potential links between the cortical actin cytoskeleton and endocytosis. Other two proteins were identified from murine macrophage cells infected with avirulent spores as ITBG-2 (CD18) and HSPA5, respectively. These data demonstrate the feasibility of using a MALDI-TOF platform to generate protein expression profiles and identify potential molecular targets for anthrax therapeutics.

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Targeting Tumor Metastasis by Regulating Nm23 Gene Expression

  • Prabhu, V. Vinod;Siddikuzzaman, Siddikuzzaman;Grace, V.M. Berlin;Guruvayoorappan, C.
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.8
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    • pp.3539-3548
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    • 2012
  • The Nm23 gene is a metastatic suppressor identified in a melanoma cell line and expressed in different tumors where their levels of expression are associated with reduced or increased metastatic potential. Nm23 is one of the over 20 metastasis suppressor genes (MSGs) confirmed in vivo. It is highly conserved from yeast to human, implying a critical developmental function. Tumors with alteration of the p53 gene and reduced expression of the Nm23 gene are more prone to metastasis. Nm23-H1 has 3'-5' exonuclease activity. This review focuses on the role of Nm23 in cancer progression and also a potential novel target for cancer therapy.

A Study on the Image-Virtualization in Fashion Illustration (패션 일러스트레이션에서의 이미지 가상화 연구)

  • Kim, Soon-Ja
    • Journal of the Korean Society of Clothing and Textiles
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    • v.32 no.3
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    • pp.505-516
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    • 2008
  • Image which appears in fashion illustration on the late twentieth century is not the representative image as an equivalence to the real fashion styles but the virtual image which bears no relation to any reality. The purpose of this study is review the concept of virtuality and analyze in which way virtual image is expressed in fashion illustrations on the background of Jean Baudrillard's simulacre theory. In post-modem paintings the expression methods of image-virtualization were image mixing through photo-image appropriation, image overlapping, and the icons inserted unreasonably, the focus-out effect through scrubbing and the over-painting on the photograph. Image-virtualization in fashion illustration was expressed through image mixing and expression of image uncertainty. Image mixing was made by photo-image appropriation, image overlapping, connection of heterogeneous images and using interface image, and uncertain image was expressed through the expression of visual ambiguity and virtual movement.

Controversies on governing the rates of protein evolution

  • Choi, Sun-Shim
    • Interdisciplinary Bio Central
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    • v.1 no.3
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    • pp.11.1-11.5
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    • 2009
  • One of the main issues of molecular evolution is to reveal the principles dictating protein evolutionary rates. A traditional hypothesis posits that protein evolutionary rates are mostly determined by the average functional importance of amino acids in a given protein. Thus the correlations of evolutionary rates with different variables such as PPI, gene essentiality and expression abundance have been studied to test the traditional hypothesis. Recently, mRNA expression abundance among the variables has drawn much attention, not only because it shows relatively strong correlation with protein evolutionary rates, but also because of the controversies surrounding an alternative hypothesis against the traditional one. Here, I will give an overview over the traditional hypothesis, and summarize the different variables that have been found to correlate with protein evolutionary rates. Then I will introduce pros and cons on the two different hypotheses.

Soluble expression and purification of synthetic human bone morphogenetic protein-2 in Escherichia coli

  • Ihm, Hyo-Jin;Yang, Seung-Ju;Huh, Jae-Wan;Choi, Soo-Young;Cho, Sung-Woo
    • BMB Reports
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    • v.41 no.5
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    • pp.404-407
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    • 2008
  • A 345-bp gene that encodes human bone morphogenetic protein-2 (hBMP-2) has been synthesized. The codon usage of the resulting gene was modified to include those triplets that are utilized in highly expressed Escherichia coli genes. The hBMP-2 gene was efficiently expressed in E. coli as a soluble and active protein. Since the recombinant hBMP-2 was readily solublized, no further solublization steps were required throughout purification. No additional tagging residues were introduced into the synthetic hBMP-2 gene product. The developed synthetic gene is a promising approach for scaling-up the soluble expression of hBMP-2.

Cloning and Expression of Escherichia coli K13 Phytase Gene (appA13) Isolated from Seawater

  • Kim Young-Ok;Kim Han-Woo;Lee Jung-Ho;Kim Kyung-Kil;Lee Jong-Yun
    • Fisheries and Aquatic Sciences
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    • v.6 no.1
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    • pp.20-26
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    • 2003
  • A bacterial strain was isolated from seawater to screen for phytase activities. A colony had the highest activity and was identified as an Escherichia coli strain. Using primers derived from E. coli acid phosphatase appA sequence, we cloned a 1,495 bp DNA fragment connected with the pGEM-T vector. It was over-expressed under lac promoter combined with its native promoter in E. coli $DH5\alpha$. The expression of the phytase gene occurred during late exponential growth and the intracellular phytase production was 16.9 units/ml. The yield of recombinant phytate was 412-fold higher than that of wild type E. coli K13.