• Title/Summary/Keyword: gene expression analysis

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Molecular Cloning and Expression of $\alpha$-Amylase Gene from Bacillus stearothermophilus in Zymomonas mobilis ZM4

  • Song, Ki-Bang
    • Journal of Microbiology and Biotechnology
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    • v.2 no.2
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    • pp.115-121
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    • 1992
  • In order to broaden the spectrum of substrate utilization of a Gram negative bacterium Zymomonas mobilis which has a great potential as an industrial ethanol producing microorganism, cloning of $\alpha$-amylase gene into Z. mobilis ZM4 was tried. The $\alpha$-amylase gene was isolated from Bacillus stearothermophilus. By Southern blot analysis, it was proven that the $\alpha$-amylase gene fragment was originated from a naturally occuring plasmid of B. stearothermophilus ATCC 31195. To place $\alpha$-amylase gene under the control of Z. mobilis promoter, two different Z. mobilis expression vectors, pZA26 and pLOI204, were used. The truncated $\alpha$-amylase gene was then introduced into these vectors. Both qualitative and quantitative activities of $\alpha$-amylase were observed in Z. mobilis cells harboring these plasmids with the $\alpha$-amylase gene inserted. Gas chromatographic analysis of ethanol showed that one of the Z. mobilis transconjugants was capable of producing 67 mM ethanol from rich medium(RM) containing 5% soluble starch as a sole carbon source.

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Rice Transformation by DNA Imbibition and Construction of Plant Vector (DNA imbibition을 이용한 벼의 형질전환과 vector 개발)

  • 유준희;남홍길정구흥
    • KSBB Journal
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    • v.8 no.2
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    • pp.104-109
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    • 1993
  • A vector for plant transformation which had two reporter genes(Gus and Hpt genes) in a single plasmid was constructed. After rice embryos imbibed DNA solution, DNA uptake and gene expression in rice were monitored. Main expression sites of the Gus gene were meristem of root and coleoptiles. There was no difference in Hpt gene expression between a single Hpt vector and the constructed vector in viability of rice in the hygromycin medium after DNA imbibition, The genomic DNA and total RNA extracted from rice transformant survived in the hygromycin medium were subjected to PCR and RT PCR analysis, respectively. As a result, we found the existence of the Hpt gene and its expression in rice.

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The Sliding Window Gene-Shaving Algorithm for Microarray Data Analysis

  • 이혜선;최대우;전치혁
    • Proceedings of the Korean Society for Bioinformatics Conference
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    • 2002.06a
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    • pp.139-152
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    • 2002
  • Gene-shaving(Hastie et al, 2000) is a very useful method to identify a meaningful group of genes when the variation of expression is large. By shaving off the low-correlated genes with the leading principal component, the primary genes with the coherent expression pattern can be identified. Gene-shaving method works well If expression levels are varied enough, but it may not catch the meaningful cluster in low expression level or different expression time even with coherent patterns. The sliding window gene-shaving method which is to apply gene-shaving in each sliding window after hierarchical clustering is to compensate losing a meaningful set of genes whose variation is not large but distinct. The performance to identify expression patterns is compared for the simulated profile data by the different variance and expression level.

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Characterization and gene expression of heat shock protein 90 in marine crab Charybdis japonica following bisphenol A and 4-nonylphenol exposures

  • Park, Kiyun;Kwak, Ihn-Sil
    • Environmental Analysis Health and Toxicology
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    • v.29
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    • pp.2.1-2.7
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    • 2014
  • Objectives Heat shock protein 90 (HSP90) is a highly conserved molecular chaperone important in the maturation of a broad spectrum of protein. In this study, an HSP90 gene was isolated from Asian paddle crab, Charybdis japonica, as a bio-indicator to monitor the marine ecosystem. Methods This work reports the responses of C. japonica HSP90 mRNA expression to cellular stress by endocrine disrupting chemicals (EDCs), such as bisphenol A (BPA) and 4-nonylphenol (NP) using real-time. reverse transcription polymerase chain reaction. Results The deduced amino acid sequence of HSP90 from C. japonica shared a high degree of homology with their homologues in other species. In a phylogenetic analysis, C. japonica HSP90 is evolutionally related with an ortholog of the other crustacean species. The expression of HSP90 gene was almost distributed in all the examined tissues of the C. japonica crab but expression levels varied among the different body parts of the crabs. We examined HSP90 mRNA expression pattern in C. japonica crabs exposed to EDCs for various exposure times. The expression of HSP90 transcripts was significantly increased in C. japonica crabs exposed to BPA and NP at different concentrations for 12, 24, 48 and 96 hours. The mRNA expression of HSP90 gene was significantly induced in a concentration- and time-dependent manner after BPA or NP exposures for 96 hours. Conclusions Taken together, expression analysis of Asian paddle crab HSP90 gene provided useful molecular information about crab responses in stress conditions and potential ways to monitor the EDCs stressors in marine environments.

A Gene Encoding Phosphatidyl Inositol-specific Phospholipase C form Cryphonectria parasitica Modulates the Hypoviral-modulated Laccase1 Expression

  • Kim, Dae-Hyuk
    • Proceedings of the Microbiological Society of Korea Conference
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    • 2005.05a
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    • pp.159-161
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    • 2005
  • Hypovirus infection of the chestnut blight fungus Cryphonectria parasitica is a useful model system to study the hypoviral regulation of fungal gene expression. The hypovirus is known to downregulate the fungal laccase1 (lac 1), the modulation of which is tightly governed by the inositol triphosphate ($IP_3$) and calcium second messenger system in a virus-free strain. We cloned the gene cplc1 encoding a phosphatidyl inositol-specific phospholipase C (PLC), in order to better characterize the fungal gene regulation by hypovirus. Sequence analysis of the cplc1 gene indicated that the protein product contained both the X and Y domains, which are the two conserved regions found in all known PLCs, with a 133 amino acid extension between the 2nd ${\beta}$-strand and the ${\alpha}$-helix in the X domain. In addition, the gene organization appeared to be highly similar to that of a ${\delta}$ type PLC. Disruption of the cplc1 gene resulted in slow growth and produced colonies characterized by little aerial mycelia and deep orange in color. In addition, down regulation of lac1 expression was observed. However, temperature sensitivity, osmosensitivity, virulence, and other hypovirulence-associated characteristics did not differ from the wild-type strain. Functional complementation of the cplc1-null mutant with the PLC1 gene from Saccharomyces cerevisiae restored lac1 expression, which suggests that the cloned gene encodes PLC activity. The present study indicates that the cplc1 gene is required for appropriate mycelial growth, and that it regulates the lac1 expression, which is also modulated by the hypovirus. Although several PLC genes have been identified in various simple eukaryotic organisms, the deletion analysis of the cplc1 gene in this study appears to be the first report on the functional analysis of PLC in filamentous fungi.

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Notch Signal Transduction Induces a Novel Profile of Kaposi's Sarcoma-Associated Herpesvirus Gene Expression

  • Chang Hee-Soon
    • Journal of Microbiology
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    • v.44 no.2
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    • pp.217-225
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    • 2006
  • Kaposi's sarcoma-associated herpesvirus (KSHV) RTA transcription factor is recruited to its responsive elements through interaction with RBP-Jk that is a downstream transcription factor of the Notch signaling pathway that is important in development and cell fate determination. This suggests that KSHV RTA mimics cellular Notch signal transduction to activate viral lytic gene expression. Here, I demonstrated that unlike other B lymphoma cells, KSHV -infected primary effusion lymphoma BCBL1 cells displayed the constitutive activation of ligand-mediated Notch signal transduction, evidenced by the Jagged ligand expression and the complete proteolytic process of Notch receptor I. In order to investigate the effect of Notch signal transduction on KSHV gene expression, human Notch intracellular (hNIC) domain that constitutively activates RBP-Jk transcription factor activity was expressed in BCBL1 cells, TRExBCBL1-hNIC, in a tetracycline inducible manner. Gene expression profiling showed that like RTA, hNIC robustly induced expression of a number of viral genes including KS immune modulatory gene resulting in downregulation of MHC I and CD54 surface expression. Finally, the genetic analysis of KSHV genome demonstrated that the hNIC-mediated expression of KS during viral latency consequently conferred the downregulation of MHC I and CD54 surface expression. These results indicate that cellular. Notch signal transduction provides a novel expression profiling of KSHV immune deregulatory gene that consequently confers the escape of host immune surveillance during viral latency.

Effect of Korean Red Ginseng through comparative analysis of cardiac gene expression in db/db mice

  • Jang, Young-Jin;Aravinthan, Adithan;Hossain, Mohammad Amjad;Kopalli, Spandana Rajendra;Kim, Bumseok;Kim, Nam Soo;Kang, Chang-Won;Kim, Jong-Hoon
    • Journal of Ginseng Research
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    • v.45 no.3
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    • pp.450-455
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    • 2021
  • Korean Red Ginseng (KRG) is an herbal oriental medicine known to alleviate cardiovascular dysfunction. To analysis the expression of diabetic cardiac complication-associated genes in db/db mice, we studied the cardiac gene expression following KRG treatment. In result, a total of 585 genes were found to be changed in db/db mice. Among the changed expression, 245 genes were found to 2-fold upregulated, and 340 genes were 2-fold downregulated. In addition, the changed gene expressions were ameliorated by KRG. In conclusion, KRG may be possible to normalize cardiac gene expressions in db/db mice.

Expression of Indica rice OsBADH1 gene under salinity stress in transgenic tobacco

  • Hasthanasombut, Supaporn;Ntui, Valentine;Supaibulwatana, Kanyaratt;Mii, Masahiro;Nakamura, Ikuo
    • Plant Biotechnology Reports
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    • v.4 no.1
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    • pp.75-83
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    • 2010
  • Glycine betaine has been reported as an osmoprotectant compound conferring tolerance to salinity and osmotic stresses in plants. We previously found that the expression of betaine aldehyde dehydrogenase 1 gene (OsBADH1), encoding a key enzyme for glycine betaine biosynthesis pathway, showed close correlation with salt tolerance of rice. In this study, the expression of the OsBADH1 gene in transgenic tobacco was investigated in response to salt stress using a transgenic approach. Transgenic tobacco plants expressing the OsBADH1 gene were generated under the control of a promoter from the maize ubiquitin gene. Three homozygous lines of $T_2$ progenies with single transgene insert were chosen for gene expression analysis. RT-PCR and western blot analysis results indicated that the OsBADH1 gene was effectively expressed in transgenic tobacco leading to the accumulation of glycine betaine. Transgenic lines demonstrated normal seed germination and morphology, and normal growth rates of seedlings under salt stress conditions. These results suggest that the OsBADH1 gene could be an excellent candidate for producing plants with osmotic stress tolerance.

Differential Display Analysis of Gene Expression Induced under DCA Treatment in Rat Liver

  • Choi, Soon-Yong;Park, Ock-Jin
    • BMB Reports
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    • v.29 no.3
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    • pp.272-275
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    • 1996
  • The expression of genes induced by Dichloroacetate (DCA) treatment was analyzed by mRNA differential display. Purified total RNAs from rat liver treated with saline or DCA (100 mg/100 g b.w.) were reverse transcribed by using a set of oligonucleotide primers. The PCR products were resolved on a denaturing sequencing gel. PCR band representing mRNA expressed specifically in DCA-treated liver was excised and reamplified by PCR. A 120-bp c-DNA clone named IC1 was isolated and the DNA sequence of IC1 was analyzed. IC1 revealed 50% homology with 3' end of a mouse fibroblast growth factor mRNA This result indicates that DCA induces the expression of a gene which has a 50% homology with a Mouse fibroblast growth factor, and expression of this gene might be involved in non genotoxic process caused by DCA.

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