• Title/Summary/Keyword: Gene expression

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Comparative Expression of Stress Related Genes in Response to Salt-stressed Aspen by Real-time RT-PCR

  • Ku, Ja-Jung;Kim, Yong-Yul
    • Korean Journal of Plant Resources
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    • v.21 no.3
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    • pp.210-215
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    • 2008
  • Gene-expression analysis is increasingly important in biological research, with real-time reverse PCR (RTPCR) becoming the method of choice for high-throughput and accurate expression profiling of selected genes. However, this technique requires important preliminary work for standardizing and optimizing the many parameters involved in the analysis. Plant stress studies are more and more based on gene expression. The analysis of gene expression requires sensitive and reproducible measurements for specific mRNA sequence. Several genes are regulated in response to abitoic stresses, such as salinity, and their gene products function in stress response and tolerance. The design of the primers and TaqMan probes for real-time PCR assays were carried out using the Primer $Express^{TM}$ software 3.0. The PCR efficiency was estimated through the linear regression of the dilution curve. To understand the expression pattern of various genes under salt stressed condition, we have developed a unique public resource of 9 stress-related genes in poplar. In this study, real-time RT-PCR was used to quantify the transcript level of 10 genes (9 stress-related genes and 1 house keeping gene) that could play a role in adaptation of Populus davidiana. Real-time RT-PCR analyses exhibited different expression ratios of related genes. The data obtained showed that determination of mRNA levels could constitute a new approach to study the stress response of P. davidiana after adaptation during growth in salinity condition.

Sequence-specific interaction between ABD-B homeodomain and castor gene in Drosophila

  • Kim, Keon-Hee;Yoo, Siuk
    • BMB Reports
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    • v.47 no.2
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    • pp.92-97
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    • 2014
  • We have examined the effect of bithorax complex genes on the expression of castor gene. During the embryonic stages 12-15, both Ultrabithorax and abdominal-A regulated the castor gene expression negatively, whereas Abdominal-B showed a positive correlation with the castor gene expression according to real-time PCR. To investigate whether ABD-B protein directly interacts with the castor gene, electrophoretic mobility shift assays were performed using the recombinant ABD-B homeodomain and oligonucleotides, which are located within the region 10 kb upstream of the castor gene. The results show that ABD-B protein directly binds to the castor gene specifically. ABD-B binds more strongly to oligonucleotides containing two 5'-TTAT-3' canonical core motifs than the probe containing the 5'-TTAC-3' motif. In addition, the sequences flanking the core motif are also involved in the protein-DNA interaction. The results demonstrate the importance of HD for direct binding to target sequences to regulate the expression level of the target genes.

Heat Inducible Expression of the CDC70 Gene Under the Control of Heat Shock Element in Saccharomyces Cerevisiae

  • Lee, Seok-Jae;Jahng, Kwang-Yeop;Lee, Young-Hoon;Chae, Keon-Sang
    • Journal of Microbiology
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    • v.33 no.3
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    • pp.196-200
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    • 1995
  • In order to express the CDC70 gene of Saccharomyces cerevisiae by heat shock, we have designed heat inducibe hybrid promoters using the Drosophila melanogaster heat shock elements (HSEs). A 220 bp-long upstream fragment of the D. melanogaster hsp70 gene comprised of four HSEs was placed upstream of the putative proximal TATA box of the CDC70 gene. Hybrid promoters containing different fusion joints were tested for their ability to drive the CDC70 gene expression by heat shock. The results showed that the HSEs of D. melanogaster conferred the heat-induced CDC70 gene expression, but the heat inducibility was much lower than that in D. melanogaster.

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Imaging Gene Expression (유전자 발현 영상기법)

  • Lee, Kyung-Han
    • The Korean Journal of Nuclear Medicine
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    • v.34 no.1
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    • pp.1-9
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    • 2000
  • The rapid progress of molecular genetic methods over the past two decades has necessitated the development of methods to detect and quantify genetic activity within living bodies. Reporter genes provide a rapid and convenient tool to monitor gene expression by yielding a readily measurable phenotype upon expression when introduced into a biological system. Conventional reporter systems, however, are limited in their usefulness for in vivo experiments or human gene therapy because of its invasive nature which requires cell damage before assays can be performed. This offers an unique opportunity for nuclear imaging techniques to develope a novel method for imaging both the location and amount of gene expression noninvasively. Current developments to achieve this goal rely on utilizing either reporter enzymes that accumulate radiolabeled substrates or reporter receptors that bind specific radioligands. This overview includes a brief introduction to the background for such research, a summary of published results, and an outlook for future directions.

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Differential gene expression by chrysotile in human bronchial epithelial cells

  • Seo, Yoo-Na;Lee, Yong-Jin;Lee, Mi-Young
    • Animal cells and systems
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    • v.16 no.2
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    • pp.95-103
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    • 2012
  • Asbestos exposure has been known to contribute to several lung diseases named asbestosis, malignant mesothelioma and lung cancer, but the disease-related molecular and cellular mechanisms are still largely unknown. To examine the effects of asbestos exposure in human bronchial epithelial cells at gene level, the global gene expression profile was analyzed following chrysotile treatment. The microarray results revealed differential gene expression in response to chrysotile treatment. The genes up- and down-regulated by chrysotile were mainly involved in processes including metabolism, signal transduction, transport, development, transcription, immune response, and other functions. The differential gene expression profiles could provide clues that might be used to understand the pathological mechanisms and therapeutic targets involved in chrysotile-related diseases.

A modified partial least squares regression for the analysis of gene expression data with survival information

  • Lee, So-Yoon;Huh, Myung-Hoe;Park, Mira
    • Journal of the Korean Data and Information Science Society
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    • v.25 no.5
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    • pp.1151-1160
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    • 2014
  • In DNA microarray studies, the number of genes far exceeds the number of samples and the gene expression measures are highly correlated. Partial least squares regression (PLSR) is one of the popular methods for dimensional reduction and known to be useful for the classifications of microarray data by several studies. In this study, we suggest a modified version of the partial least squares regression to analyze gene expression data with survival information. The method is designed as a new gene selection method using PLSR with an iterative procedure of imputing censored survival time. Mean square error of prediction criterion is used to determine the dimension of the model. To visualize the data, plot for variables superimposed with samples are used. The method is applied to two microarray data sets, both containing survival time. The results show that the proposed method works well for interpreting gene expression microarray data.

Bcl-2 Gene Expression in Human Breast Cancers in Iran

  • Rostamizadeh, Leila;Fakhrjou, Ashraf;Montazeri, Vahid;Estiar, Mehrdad Asghari;Naghavi-Behzad, Mohammad;Hosseini, Somayyeh;Sakhinia, Masoud;Sakhinia, Ebrahim
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.7
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    • pp.4209-4214
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    • 2013
  • Background: Breast cancer is among the five most common cancers and ranks first among cancers diagnosed in Iranian women. Screening and treatment of this disease with molecular methods, especially regarding high incidences at early age and advanced stage, is essential. Several genes with altered expression have been identified by cDNA microarray studies in breast cancer, with the Bcl-2 gene indicated as a likely candidate. In this study, we studied Bcl-2 gene expression levels in parallel tumor and non-tumor breast tissues. Materials and Methods: Forty samples including 21 tumor, 16 non tumor (marginal) and 3 benign breast tissues which were all pathologically diagnosed, were subjected to RNA extraction and polyA RT-PCR with the expression level of Bcl-2 quantified using real-time PCR. Results: There is higher expression levels of the Bcl-2 gene in tumor samples compared with marginal samples, but not attaining significance(p>0.05). Bcl-2 expression in 14 (66.7%) of the cases of tumor samples and 9 (56.3%) cases of the marginal samples were positive. Comparison of the expression of the Bcl-2 gene in histological grade showed that a high expression of Bcl-2 was associated with a high histological grade (p<0.41). Conclusions: Our data suggests that dysregulated Bcl-2 gene expression is potentially involved in the pathogenesis of breast cancer. Using gene expression analysis may significantly improve our ability for screening cancer patients and will prove a powerful tool in the diagnosis and prognostic evaluation of the disease whilst aiding the cooperative group trials in the Bcl-2 based therapy project.

Statistical Tests for Time Course Microarray Experiments

  • Park, Tae-Seong;Lee, Seong-Gon;Choe, Ho-Sik;Lee, Seung-Yeon;Lee, Yong-Seong
    • Proceedings of the Korean Statistical Society Conference
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    • 2002.05a
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    • pp.85-90
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    • 2002
  • Microarray technology allows the monitoring of expression levels for thousands of genes simultaneously. In time-course experiments in which gene expression is monitored over time we are interested in testing gene expression profiles for different experimental groups. We propose a statistical test based on the ANOVA model to identify genes that have different gene expression profiles among experimental groups in time-course experiments. Using this test, we can detect genes that have different gene expression profiles among experimental groups. The proposed model is illustrated using cDNA microarrays of 3,840 genes obtained in an experiment to search for changes in gene expression profiles during neuronal differentiation of cortical stem cells.

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Consensus Clustering for Time Course Gene Expression Microarray Data

  • Kim, Seo-Young;Bae, Jong-Sung
    • Communications for Statistical Applications and Methods
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    • v.12 no.2
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    • pp.335-348
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    • 2005
  • The rapid development of microarray technologies enabled the monitoring of expression levels of thousands of genes simultaneously. Recently, the time course gene expression data are often measured to study dynamic biological systems and gene regulatory networks. For the data, biologists are attempting to group genes based on the temporal pattern of their expression levels. We apply the consensus clustering algorithm to a time course gene expression data in order to infer statistically meaningful information from the measurements. We evaluate each of consensus clustering and existing clustering methods with various validation measures. In this paper, we consider hierarchical clustering and Diana of existing methods, and consensus clustering with hierarchical clustering, Diana and mixed hierachical and Diana methods and evaluate their performances on a real micro array data set and two simulated data sets.

Construction of Yeast Vectors Potentially Useful for Expression of Eukaryotic Genes as ${\beta}$-galactosidase Fusion Proteins

  • Chung, Kyung-Sook;Choi, Won-Ja;Lee, Hee-Won;Kim, Kyu-Won;Yoo, Hyang-Sook
    • BMB Reports
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    • v.29 no.4
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    • pp.359-364
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    • 1996
  • By both in vitro hydroxylamine mutagenesis of the wild type 3-phosphoglycerate kinase gene (PGK) promoter DNA and insertion of the leu2-d gene, we have created yeast expression vectors potentially useful for production of eukaryotic genes in yeast. The guanine (G) to adenine (A) change at the -3 position from the ATG start codon of the PGK promoter-based vector rendered a 6~7 times elevated expression of the adjacent eukaryotic gene, and insertion of the leu2-d gene in the vector containing the mutated PGK promoter further enhanced the expression of the gene. When expression of the AIDS virus HIV1-gagP17 gene in a lacZ fusion form was examined with this new vector, a 15 times higher level of expression than that from the original PGK promoter was observed. Northern and Southern analysis showed that this elevated expression is due to the production of a high copy number of mRNA by leu2-d gene functioning and by efficient translation of the produced mRNA. Thus, the vector that contained the A at the -3 position from the ATG start codon in the promoter region and the leu2-d gene shows increased expression capability and will be potentially useful for production of eukaryotic genes in yeast.

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