• Title/Summary/Keyword: Promoter analysis

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A Numerical Study on Performance of Air-to-Air Plastic Plate Heat Exchanger

  • Chung, Min-Ho;Yoo, Seong-Yeon;Han, Kyu-Hyun;Yoon, Hong-Ik;Kang, Hyoung-Chul
    • International Journal of Air-Conditioning and Refrigeration
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    • v.17 no.2
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    • pp.52-60
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    • 2009
  • The purpose of this research is to develop high efficiency plastic plate heat exchangers which can be substituted for conventional aluminum plate heat exchangers. Four simulation models of plastic plate heat exchangers are designed and simulated: that is, flat plate type, turbulent promoter type, corrugate type and dimple type heat exchanger. The flat plate type is designed as the reference model in order to evaluate how much thermal performance increases. The turbulent promoter type is fabricated with cylindrical-type vortex generators and rib-type turbulent promoters. The corrugate type is obtained from the conventional stainless steel compact heat exchangers, which are called the herringbone-type compact heat exchangers. The dimple type has a number of dimples on its surface. In this study, the flow and heat transfer characteristics of the plastic plate heat exchanger are investigated using numerical simulation and compared with experimental results. Numerical simulation is carried out using the FLUENT code. The flows are assumed as a three-dimensional, incompressible and turbulent model. The computational analysis and experimental results both show that the friction coefficient and Nu number is highest in the corrugate type. The tendency of numerical simulation results is in good agreement with that of the experimental results.

Structure and expression analysis of the OsCam1-1 calmodulin gene from Oryza sativa L.

  • Phean-o-pas, Srivilai;Limpaseni, Tipaporn;Buaboocha, Teerapong
    • BMB Reports
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    • v.41 no.11
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    • pp.771-777
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    • 2008
  • Calmodulin (CaM) proteins, members of the EF-hand family of $Ca^{2+}$-binding proteins, represent important relays in plant calcium signals. Here, OsCam1-1 was isolated by PCR amplification from the rice genome. The gene contains an ORF of 450 base pairs with a single intron at the same position found in other plant Cam genes. A promoter region with a TATA box at position-26 was predicted and fused to a gus reporter gene, and this construct was used to produce transgenic rice by Agrobacterium-mediated transformation. GUS activity was observed in all organs examined and throughout tissues in cross-sections, but activity was strongest in the vascular bundles of leaves and the vascular cylinders of roots. To examine the properties of OsCaM1-1, the encoding cDNA was expressed in Escherichia coli. The electrophoretic mobility shift when incubated with $Ca^{2+}$ indicates that recombinant OsCaM1-1 is a functional $Ca^{2+}$-binding protein. In addition, OsCaM1-1 bound the CaMKII target peptide confirming its likely functionality as a calmodulin.

Role of Intergenic and 3'-Proximal Noncoding Regions in Coat Protein Expression and Replication of Barley yellow dwarf virus PAV

  • Moon, Jae-Sun;Nancy K. McCoppin;Leslie L. Domier
    • The Plant Pathology Journal
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    • v.17 no.1
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    • pp.22-28
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    • 2001
  • Barley yellow dwarf virus PAV (BYDV-PAV) has a 5.7-kb positive-sense single-stranded RNA genome that contains six open reading frames (ORFs). BYDV-PAV produces three subgenomic RNAs (sgRNAs). The largest of which encodes the coat, 17-kDa, and readthrough proteins from two initiation codons. To investigate the role of intergenic and 3'-proximal noncoding regions (NCRs) in coat protein (CP) expression and BYDV-PAV replication, a full-length infectious cDNA of the RNA genome of an Illinois isolate of BYDV-PAV was constructed downstream of the Cauliflower mosaic virus-35S promoter. Linear DNA molecules of these cDNAs were infectious, expressed the 22-kDa CP, and produced both genomic RNA sgRNAs in ratios similar to those observed in protoplasts inoculated with viral RNA. The portion of 5'NCR of sgRNA1 between ORFs 2 and 3 was not required for, but enhanced translation of CP from ORF3. Mutants containing deletions in the NCR downstream of ORF5 failed to replicate in oat protoplasts. These results indicate that an intact 3$^1$NCR is required for BYDV-PAV replication.

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Expression of resveratrol synthase gene and accumulation of resveratrol in transgenic potatoes (Solanum tuberosum L.)

  • Yi, Jung Yoon;Seo, Hyo Won;Yun, Song Joong;Ok, HyunChoong;Park, YoungEun;Cho, Ji Hong;Cho, HyunMook
    • Korean Journal of Breeding Science
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    • v.41 no.4
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    • pp.385-390
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    • 2009
  • A resveratrol synthase (RS) gene was isolated from peanut (Arachis hypogaea, L. cv. Jinpoong) plants. This gene was placed under the control of the cauliflower mosaic virus 35S promoter (CaMV35S) and introduced into two Korean varieties of potato (Solanum tuberosum L. cvs. Jasim and Jowon) plants by Agrobacterium-mediated gene transfer. Putative transformants were screened by PCR with primers designed from CaMV 35S promoter, NOS terminator and RS gene. Most of selected transgenic potato plants showed the amplification of expected fragments by PCR of genomic DNA with gene-specific primers, while they were absent in untransformed control plants. Expression of the resveratrol synthase gene was also examined by northern blot analysis. The transformants showed a band which was lacking in the control plant, confirming that the introduced gene is transcribed into mRNA in the transformants. The strength of the band, which reflected the level of mRNA expression, differed among the individual transformants. Among the transformants obtained, the highest trans-resveratrol content in the transgenic young leaves of purple-fleshed "Jashim" was $2.11{\mu}gg^{-1}$ fresh weight and that in the microtubers in vitro of purple fleshed "Jashim" was $8.31{\mu}gg^{-1}$ fresh weight. This amount of resveratrol may have a positive biological effect on human health.

Solving the Mystery of Consistent Negative/Low Net Promoter Score (NPS) in Cross-Cultural Marketing Research

  • Seth, Sanjay;Scott, Don;Svihel, Chad;Murphy-Shigematsu, Stephen
    • Asia Marketing Journal
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    • v.17 no.4
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    • pp.43-61
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    • 2016
  • This paper has identified some theoretical reasons and empirical evidence for negative scores that occur in Japan and Korea or unstable NPS scores that can be experienced. A psychological analysis of NPS results sheds light on the validity of the negative NPS scores that are often found in Japan and Korea. Usually customer experience surveys utilize a "single stimulus" such as the "company" or the "company's products / services." However, in the case of the "recommendation to friend" question of the NPS system there are two stimuli namely the "company product/service" and the influence of "friends." Hence, the survey outcomes from this question can be very different when compared with other single stimulus questions such as "overall satisfaction" or "repurchase." Japanese and Korean people may have a positive attitude towards the company but they will provide low NPS scores because they are reflecting that they would not run the risk of ruining their relationships with their friends by making a recommendation. As a result, in the NPS system these people will be labeled as "detractors" when in fact they are "ambivalent customers." Using several Japanese and Korean based marketing research industry examples and case studies, different strategies are proposed to address the issue of negative scores in the NPS system in Japan and Korea. The Customers Psyche appears to be the key determinant factors for both types of behavioural items (items with a single stimulus as well as items with two stimuli).

Vorinostat-induced acetylation of RUNX3 reshapes transcriptional profile through long-range enhancer-promoter interactions in natural killer cells

  • Eun-Chong Lee;Kyungwoo Kim;Woong-Jae Jung;Hyoung-Pyo Kim
    • BMB Reports
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    • v.56 no.7
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    • pp.398-403
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    • 2023
  • Natural killer (NK) cells are an essential part of the innate immune system that helps control infections and tumors. Recent studies have shown that Vorinostat, a histone deacetylase (HDAC) inhibitor, can cause significant changes in gene expression and signaling pathways in NK cells. Since gene expression in eukaryotic cells is closely linked to the complex three-dimensional (3D) chromatin architecture, an integrative analysis of the transcriptome, histone profiling, chromatin accessibility, and 3D genome organization is needed to gain a more comprehensive understanding of how Vorinostat impacts transcription regulation of NK cells from a chromatin-based perspective. The results demonstrate that Vorinostat treatment reprograms the enhancer landscapes of the human NK-92 NK cell line while overall 3D genome organization remains largely stable. Moreover, we identified that the Vorinostat-induced RUNX3 acetylation is linked to the increased enhancer activity, leading to elevated expression of immune response-related genes via long-range enhancer-promoter chromatin interactions. In summary, these findings have important implications in the development of new therapies for cancer and immune-related diseases by shedding light on the mechanisms underlying Vorinostat's impact on transcriptional regulation in NK cells within the context of 3D enhancer network.

Tumor antigen PRAME is a potential therapeutic target of p53 activation in melanoma cells

  • Yong-Kyu Lee;Hyeon Ho Heo;Nackhyoung Kim;Ui-Hyun Park;Hyesook Youn;Eun-Yi Moon;Eun-Joo Kim;Soo-Jong Um
    • BMB Reports
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    • v.57 no.6
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    • pp.299-304
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    • 2024
  • Upregulation of PRAME (preferentially expressed antigen of melanoma) has been implicated in the progression of a variety of cancers, including melanoma. The tumor suppressor p53 is a transcriptional regulator that mediates cell cycle arrest and apoptosis in response to stress signals. Here, we report that PRAME is a novel repressive target of p53. This was supported by analysis of melanoma cell lines carrying wild-type p53 and human melanoma databases. mRNA expression of PRAME was downregulated by p53 overexpression and activation using DNA-damaging agents, but upregulated by p53 depletion. We identified a p53-responsive element (p53RE) in the promoter region of PRAME. Luciferase and ChIP assays showed that p53 represses the transcriptional activity of the PRAME promoter and is recruited to the p53RE together with HDAC1 upon etoposide treatment. The functional significance of p53 activation-mediated PRAME downregulation was demonstrated by measuring colony formation and p27 expression in melanoma cells. These data suggest that p53 activation, which leads to PRAME downregulation, could be a therapeutic strategy in melanoma cells.

Resveratrol epigenetically regulates the expression of zinc finger protein 36 in non-small cell lung cancer cell lines

  • Ahmad Fudhaili;Nal Ae Yoon;Seokmin Kang;Jinhyun Ryu;Joo Yeon Jeong;Dong Hoon Lee;Sang Soo Kang
    • Oncology Reports
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    • v.41 no.2
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    • pp.1377-1386
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    • 2019
  • Zinc finger protein 36 (ZFP36) is an AU-rich element protein that binds to 3'-untranslated regions and promotes the decay of target mRNAs. Downregulation of ZFP36 expression in turn results in stabilization of target mRNAs. A recent study indicated that downregulation of ZFP36 expression in human liver cancer is caused by epigenetic mechanisms. The purpose of the present study was to investigate the potential of resveratrol (Res) to induce ZFP36 expression. Promoter methylation was analyzed using methylation-sensitive restriction analysis. It was determined that Res treatment increased ZFP36 expression and decreased the mRNA levels of ZFP36 target genes in A549 lung cancer cells. Additionally, Res suppressed the expression of DNA (cytosine-5)-methyltransferase 1 and induced demethylation of the ZFP36 promoter. Collectively, the present results demonstrated that Res has anticancer activity through its epigenetic regulation of ZFP36 in non-small cell lung cancer.

Production of porcine fibroblasts carrying a vector enforced specific expression of CD73 to endothelial cells (돼지 혈관내피세포 특이적 CD73 발현 벡터가 도입된 돼지 섬유아세포 생산)

  • Oh, Keon Bong;Lee, Haesun;Hwang, Seongsoo;Ock, Sun-A;Chung, Hak-Jae;Byun, Sung June;Lee, Poongyeon;Im, Gi-Sun
    • Journal of Embryo Transfer
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    • v.31 no.3
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    • pp.161-168
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    • 2016
  • Nucleotide metabolism in endothelium is variable between different species. Recent studies demonstrated that this variability could contribute coagulation dysfunction, even though organs of the alpha 1,3-galactosyltransferase gene knockout pig were transplanted into the primate. CD73 (ecto-5'-nucelotidase) is an enzyme at cell surface catalyzing the hydrolysis of adenosine triphosphate to adenosine, which plays role on a substance for anti-inflammatory and anti-coagulant. Thus, overexpression of CD73 in endothelial cells of the pig is considered as an approach to reduce coagulopathy. In this study, we constructed a human CD73 expression vector under control of porcine Icam2 promoter (pIcam2-hCD73), which is expressed specifically at endothelial cells, and of CMV promoter as a control (CMV-CD73). First, we transfected the CMV-CD73 vector into HEK293 cells, and then confirmed CD73 expression at cell surface by flow cytometry analysis. Next, we transfected the pIcma2-CD73 and CMV-CD73 vectors into primary porcine fibroblasts and endothelial cells. Consequence was that the pIcma2-CD73 vector was expressed only at the porcine endothelial cells, meaning that the pIcam2 promoter lead to endothelial cell-specific expression of CD73 in vitro. Finally, we nucleofected the pIcam2-hCD73 vector into passage 3 fibroblasts, and enforced hygromycin selection of 400mg/ml. We were able to obtain forty three colonies harboring pIcam2-CD73 to provide donor cells for transgenic cloned porcine production.

Improvement of production of active cyclodextrin glucanotransferase by coexpression GroEL/ES chaperons in E. coli (E. coli에서 GroEL/ES chaperone 공발현에 의한 활성형 cyclodextrin glucanotransferase의 생산 증대)

  • 권미정;박소림;김병우;김성구;남수완
    • Journal of Life Science
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    • v.12 no.6
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    • pp.688-693
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    • 2002
  • Molecular chaperones prevent the misfolding of newly synthesized polypeptides in the cell. The coexpression of molecular chaperones could be expected to improve the production of soluble and active recombinant proteins. In this study, the effect of coexpression of E. coli GroEL/ES chaperone on the active production of Bacillus macerans cyclodextrin glucanotransferase (CGTase) in E. coli was investigated. Two plasmids, pTCGT1 and pGro7 in which the cgt and the groEL/ES genes are under the control of 77 promoter and araB promoter, respectively, were co-transformed into E. coli. With a series of cultures of recombinant E. coli cells, the optimal concentrations of IPTG and L-arabinose were found be 1 mM and 0.3 mg/$m\ell$, respectively. When IPTG and L-arabinose were added at 0.8~1.0 $OD_{600}$ and 0.4~0.5 $OD_{600}$, active CGTase production was increased significantly. This coexpression condition resulted in 1.5-fold increased level of soluble CGTase (0.7~0.73 unit/$m\ell$), compared to the level of CGTase in the single expression (0.36~0.56 unit/$m\ell$). An SDS-PACE analysis revealed that about 33.6% of CGTase in the total CGTase protein was found in the soluble fraction by coexpression of GroEL/ES chaperone.