• Title/Summary/Keyword: transcriptional analysis

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Homologous Expression and Quantitative Analysis of T3SS-Dependent Secretion of TAP-Tagged XoAvrBs2 in Xanthomonas oryzae pv. oryzae Induced by Rice Leaf Extract

  • Kim, S.H.;Lee, S.E.;Hong, M.K.;Song, N.H.;Yoon, B.;Viet, P.T.;Ahn, Y.J.;Lee, B.M.;Jung, J.W.;Kim, K.P.;Han, Y.S.;Kim, J.G.;Kang, L.W.
    • Journal of Microbiology and Biotechnology
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    • v.21 no.7
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    • pp.679-685
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    • 2011
  • Xanthomonas oryzae pv. oryzae (Xoo) produces a putative effector, XoAvrBs2. We expressed XoAvrBs2 homologously in Xoo with a TAP-tag at the C-terminus to enable quantitative analysis of protein expression and secretion. Addition of rice leaf extracts from both Xoo-sensitive and Xoo-resistant rice cultivars to the Xoo cells induced expression of the XoAvrBs2 gene at the transcriptional and translational levels, and also stimulated a remarkable amount of XoAvrBs2 secretion into the medium. In a T3SS-defective Xoo mutant strain, secretion of the TAPtagged XoAvrBs2 was blocked. Thus, we elucidated the transcriptional and translational expressions of the XoAvrBs2 gene in Xoo was induced in vitro by the interaction with rice and the induced secretion of XoAvrBs2 was T3SSdependent. It is the first report to measure the homologous expression and secretion of XoAvrBs2 in vitro by rice leaf extract. Our system for the quantitative analysis of effector protein expression and secretion could be generally used for the study of host-pathogen interactions.

Transcriptome profiling and identification of functional genes involved in H2S response in grapevine tissue cultured plantlets

  • Ma, Qian;Yang, Jingli
    • Genes and Genomics
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    • v.40 no.12
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    • pp.1287-1300
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    • 2018
  • Hydrogen sulfide ($H_2S$), a small bioactive gas, has been proved functioning in plant growth and development as well as alleviation of abiotic stresses, which including promoting seed germination, accelerating embryonic root growth, regulating flower senescence, inducing stomatal closure, and defending drought, heat, heavy metals and osmotic stresses etc. However, the molecular functioning mechanism of $H_2S$ was still unclear. The primary objective of this research was to analyze the transcriptional differences and functional genes involved in the $H_2S$ responses. In details, 4-week-old plantlets in tissue culture of grapevine (Vitis vinifera L.) cultivar 'Zuoyouhong' were sprayed with 0.1 mM NaHS for 12 h, and then transcriptome sequencing and qRT-PCR analysis were used to study the transcriptional differences and functional genes involved in the $H_2S$ responses. Our results indicated that 650 genes were differentially expressed after $H_2S$ treatment, in which 224 genes were up-regulated and 426 genes were down-regulated. The GO enrichment analysis and KEGG enrichment analysis results indicated that the up-regulated genes after $H_2S$ treatment focused on carbon metabolism, biosynthesis of amino acids, and glycolysis/gluconeogenesis, and the down-regulated genes were mainly in metabolic pathways, biosynthesis of secondary metabolites, and plant hormone signal transduction. Analyzing the transcription factor coding genes in details, it was indicated that 10 AP2/EREBPs, 5 NACs, 3 WRKYs, 3 MYBs, and 2 bHLHs etc. transcription factor coding genes were up-regulated, while 4 MYBs, 3 OFPs, 3 bHLHs, 2 AP2/EREBPs, 2 HBs etc. transcription factor coding genes were down-regulated. Taken together, $H_2S$ increased the productions in secondary metabolites and a variety of defensive compounds to improve plant development and abiotic resistance, and extend fruits postharvest shelf life by regulating the expression of AP2/EREBPs, WRKYs, MYBs, CABs, GRIP22, FERRITINs, TPSs, UGTs, and GHs etc.

Control of Tylosin Biosynthesis in Streptomyces fradiae

  • Cundliffe, Eric
    • Journal of Microbiology and Biotechnology
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    • v.18 no.9
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    • pp.1485-1491
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    • 2008
  • Tylosin biosynthesis is controlled in cascade fashion by multiple transcriptional regulators, acting positively or negatively, in conjunction with a signalling ligand that acts as a classical inducer. The roles of regulatory gene products have been characterized by a combination of gene expression analysis and fermentation studies, using engineered strains of S. fradiae in which specific genes were inactivated or overexpressed. Among various novel features of the regulatory model, involvement of the signalling ligand is not essential for tylosin biosynthesis.

Genetic Regulation of Corynebacterium glutamicum Metabolism

  • Wendisch Volker F.
    • Journal of Microbiology and Biotechnology
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    • v.16 no.7
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    • pp.999-1009
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    • 2006
  • Physiological, biochemical and genetic studies of Corynebacterium glutamicum, a workhorse of white biotechnology used for amino acid production, led to a waste knowledge mainly about amino acid biosynthetic pathways and the central carbon metabolism of this bacterium. Spurred by the availability of the genome sequence and of genome-based experimental methods such as DNA microarray analysis, research on genetic regulation came into focus. Recent progress on mechanisms of genetic regulation of the carbon, nitrogen, sulfur and phosphorus metabolism in C. glutamicum will be discussed.

Transcriptional Responses of Respiratory Epithelial Cells to Nontypable H. influenzae Infection: Identification of Differentially Regulated Genes by Microarray Analysis of Human cDNA

  • Lee, Na-Gyong
    • Proceedings of the PSK Conference
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    • 2002.10a
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    • pp.151-152
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    • 2002
  • Bacterial infection is a very complex process in which both pathogenic microorganisms and host cells play crucial roles, and it is the outcome of interactions between the two participants. To elucidate the bacterial pathogenesis mechanisms, therefore, it is essential to understand the cellular and systemic responses of the host as well as the virulence factors of the pathogen. Infection of a host by pathogenic bacteria causes drastic changes in the physiology of host cells, leading to activation of a program of various gene expression. (omitted)

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Characterization of PpsR, a Transcriptional Repressor of the Expression of Photosysem Gene, from Rhodobacter sphaeroides

  • Cho, Seung-Hyun;Kang, Sa-Ouk
    • Proceedings of the Korean Biophysical Society Conference
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    • 2002.06b
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    • pp.51-51
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    • 2002
  • PpsR from the facultative photohetrotroph Rhodobacter sphaeroides is involved in repression of photo system gene expression. SDS-PAGE analysis showed that some portion of PpsR is oxidized so that intra- or inter-disulfide bond is formed between the two cysteins in each subunit. The disulfide bond was reduced by dithiothreitol and the binding activity to puc promoter region was increased.(omitted)

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Optimized Internal Control and Gene Expression Analysis in Epstein-Barr Virus-Transformed Lymphoblastoid Cell Lines

  • Nam, Hye-Young;Kim, Hye-Ryun;Shim, Sung-Mi;Lee, Jae-Eun;Kim, Jun-Woo;Park, Hye-Kyung;Han, Bok-Ghee;Jeon, Jae-Pil
    • Genomics & Informatics
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    • v.9 no.3
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    • pp.127-133
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    • 2011
  • The Epstein-Barr virus-transformed lymphoblastoid cell line (LCL) is one of the major genomic resources for human genetics and immunological studies. Use of LCLs is currently extended to pharmacogenetic studies to investigate variations in human gene expression as well as drug responses between individuals. We evaluated four common internal controls for gene expression analysis of selected hematopoietic transcriptional regulatory genes between B cells and LCLs. In this study, the expression pattern analyses showed that TBP (TATA box-binding protein) is a suitable internal control for normalization, whereas GAPDH (glyceraldehyde-3-phosphate dehydrogenase) is not a good internal control for gene expression analyses of hematopoiesis-related genes between B cells and LCLs at different subculture passages. Using the TBP normalizer, we found significant gene expression changes in selected hematopoietic transcriptional regulatory genes (downregulation of RUNX1, RUNX3, CBFB, TLE1, and NOTCH2 ; upregulation of MSC and PLAGL2) between B cells and LCLs at different passage numbers. These results suggest that these hematopoietic transcriptional regulatory genes are potential cellular targets of EBV infection, contributing to EBV-mediated B-cell transformation and LCL immortalization.

ZNF552, a novel human KRAB/C2H2 zinc finger protein, inhibits AP-1- and SRE-mediated transcriptional activity

  • Deng, Yun;Liu, Bisheng;Fan, Xiongwei;Wang, Yuequn;Tang, Ming;Mo, Xiaoyang;Li, Yongqing;Ying, Zaochu;Wan, Yongqi;Luo, Na;Zhou, Junmei;Wu, Xiushan;Yuan, Wuzhou
    • BMB Reports
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    • v.43 no.3
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    • pp.193-198
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    • 2010
  • In this study, we report the identification and characterization of a novel C2H2 zinc finger protein, ZNF552, from a human embryonic heart cDNA library. ZNF552 is composed of three exons and two introns and maps to chromosome 19q13.43. The cDNA of ZNF552 is 2.3 kb, encoding 407 amino acids with an amino-terminal KRAB domain and seven carboxyl-terminal C2H2 zinc finger motifs in the nucleus and cytoplasm. Northern blotting analysis indicated that a 2.3 kb transcript specific for ZNF552 was expressed in liver, lung, spleen, testis and kidney, especially with a higher level in the lung and testis in human adult tissues. Reporter gene assays showed that ZNF552 was a transcriptional repressor, and overexpression of ZNF552 in the COS-7 cells inhibited the transcriptional activities of AP-1 and SRE, which could be relieved through RNAi analysis. Deletion studies showed that the KRAB domain of ZNF552 may be involved in this inhibition.

Expression Analysis of the csp-like Genes from Corynebacterium glutamicum Encoding Homologs of the Escherichia coli Major Cold-Shock Protein CspA

  • Kim, Wan-Soo;Park, Soo-Dong;Lee, Seok-Myung;Kim, Youn-Hee;Kim, Pil;Lee, Heung-Shick
    • Journal of Microbiology and Biotechnology
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    • v.17 no.8
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    • pp.1353-1360
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    • 2007
  • Three csp-like genes were identified in the Corynebacterium glutamicum genome and designated cspA, cspB, and cspA2. The genes cspA and cspA2 encode proteins, comprising of 67 amino acid residues, respectively. They share 83% identity with each other. Identity of those proteins with Escherichia coli Csp proteins was near 50%. The cspB gene encodes a protein composed of 127 amino acids, which has 40% and 35% sequence identity with CspA and CspA2, respectively, especially at its N-terminal region. Analysis of the gene expression profiles was done using transcriptional cat fusion, which identified not only active expression of the three genes at the physiological growth temperature of $30^{\circ}C$ but also growth phase-dependent expression with the highest activity at late log phase. The promoters of cspA and cspA2 were more active than that of cspB. The expression of the two genes increased by 30% after a temperature downshift to $15^{\circ}C$, and such stimulation was more evident in the late growth phase. In addition, the cspA gene appeared to show DNA-binding activity in vivo, and the activity increased at lower temperatures. Interestingly, the presence of cspA in multicopy hindered the growth of the host C. glutamicum cells at $20^{\circ}C$, but not at $30^{\circ}C$. Altogether, these data suggest that cspA, cspB, and cspA2 perform functions related to cold shock as well as normal cellular physiology. Moreover, CspA and its ortholog CspA2 may perform additional functions as a transcriptional regulator.