• Title/Summary/Keyword: Promoter strength

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Adhesive Properties of Epoxy Composite According to the Surface Treatment of Cu Substrate and Adhesion Promoter Content (구리기판의 표면처리 및 접착증진제 함량에 따른 에폭시 컴포지트의 접착특성)

  • Eun-jin Kim;Jung Soo Kim;Young-Wook Chang;Dong Hyun Kim
    • Journal of Adhesion and Interface
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    • v.23 no.4
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    • pp.108-115
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    • 2022
  • In this study, we synthesized poly(itaconic acid-co-acrylamide) (IAcAAM) used as a novel polymer adhesion promoter to improve the adhesion strength of surface-treated Cu lead frames and epoxy composites. IAcAAM comprising itaconic acid, acrylamide was prepared through radical aqueous polymerization. The chemical structure and properties of IAcAAM was analyzed by FT-IR, 1H-NMR, GPC, and DSC. The surface of the copper lead frame was treated with high temperature, alkali, and UV ozone to reduce the water contact angle and increase the surface energy. The adhesive strength of Cu lead frame and epoxy composite increased with the decrease of contact angle. The adhesive strength of Cu lead frame/epoxy composite increased with the addition of IAcAAM in epoxy composite. As silica content increased, the adhesive strength of Cu lead frame and epoxy composite tended to slightly decrease.

Expression of Inulinase Gene by Yeast Constitutive Promoters (효모의 구성적 Promoter들에 의한 Inulinase 유전자의 발현)

  • 김연희;남수완
    • Journal of Life Science
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    • v.9 no.2
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    • pp.153-159
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    • 1999
  • To express constitutively the inulinase gene (INUl) of Kluyveromyces marxianus in Saccharomyces cerevisiae, three yeast promoters such as GAPDH, ADH1 and ENO1 were connected upstream of INUl. The resulting plasmids, pYIGP, pADHl-INU, and pENO-INU were introduced to S. cerevisiae SEY2102 host strain, respectively, and then each transformants were selected by staining of colonies on sucrose-agar plate. When the yeast transformants were cultivated on 2$\%$ dextrose media, the total expression levels of inulinase reached to 1.11 unit/mL, 0.88 unit/mL, and 0.69 unit/mL for respective GAPDH, ADH1, and ENO1 promoter systems. On 4% dextrose media, however, the inulinase activities were observed at 2.00 unit/mL for pYIGP, 0.71 unit/mL for pADH1-INU, and 1.40 unit/mL for pENO-INU. This result indicates that the constitutive expression of INUl was significantly affected by the initial concentration of dextrose and the promoter strength was in the order GAPDH, ENO1, and ADH1 promoter at high dextrose concentration. Taking into account the plasmid stability, however, it is suggested that the ENO1 promoter system is more suitable for the INU1 expression on high dextrose medium or in the fed-batch cultivation accumulating ethanol at high level.

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Deletion Analysis of Pichia PGK1 Promoter and Construction of an Episomal Vector for Heterologous Protein Expression in P. pastoris (Pichia PGK1프로모터의 분석과 P. pastoris에 있어 외래단백질발현을 위한 Episomal벡터의 제조)

  • Lee, Sung-Jae;Hong, In-Pyo;Baek, Seon-Yeol;Choi, Shin-Geon
    • Microbiology and Biotechnology Letters
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    • v.35 no.3
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    • pp.184-190
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    • 2007
  • Approximately 2.0 kb of the promoter region of the Pichia pastoris phosphoglycerate kinase gene (PGK1) was reduced to a 266 bp fragment and this minimized portion was used for construction of a new episomal constitutive expression vector in P. pastoris. As an approach to developing a constitutive expression vector in P. pastoris, the GAP promoter region of the Pichia expression vector pGAPZB was replaced with sequentially deleted PGK1 promoter fragments fused to a beta-galactosidase gene. When a lacZ gene was used as a reporter gene, PGK1 promoter strength was lower than that of the constitutive GAP promoter but it was higher than TEF1. We report here the development of the pPGKZ-E vector as a new episomal expression vector for heterologous gene expression by removing non-essential regions of the PGK1 promoter. This broadens the choice of episomal expression vectors for controlled constitutive expression in P. pastoris.

Association between p16 Promoter Methylation and Thyroid Cancer Risk: A Meta-analysis

  • Wu, Wei;Yang, Sheng-Fu;Liu, Fei-Fei;Zhang, Ji-Hong
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.16
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    • pp.7111-7115
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    • 2015
  • Background: The aim of the meta-analysis was to derive a more precise assessment of the association between p16 promoter methylation and thyroid cancer risk. Materials and Methods: The PubMed, Web of Science databases and Chinese CNKI were searched for relevant articles. Ultimately, seventeen case-control studies were included with a total of 804 thyroid cancer cases and 487 controls analysis by R Software (R version 3.1.2) including meta. Crude odds ratios with 95% confidence intervals were calculated using the random-effects model which were used to assess the strength of relationship between p16 methylation and lung carcinogenesis. Funnel plots were carried out to evaluate publication bias. Results: The meta-analysis results showed that the frequency of p16 promoter methylation in cancer tissue/blood was significantly higher than that normal tissue/blood (OR=5.46, 95%CI 3.12-9.55, P<0.0001) by random effects model with small heterogeneity. Conclusions: Thus, p16 promoter methylation may be associated with thyroid cancer risk.

Development of Enhanced Yeast Expression System for GAP Promoter by Directed Evolution

  • Kang, Whan-Koo;Hwang, Sun-Duk;Kim, Bum-Chang;Lee, Chul-Woo;Son, Jeong-Il;Kim, Hyoung-Sik;Lee, Byung-Ryul;Lee, Bheong-Uk
    • 한국생물공학회:학술대회논문집
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    • 2003.10a
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    • pp.753-757
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    • 2003
  • Escherichia coli and Saccharomyces cerevisiae have been used as host for production of recombinant proteins. It is known that S. cerevisiae has advantages such as good folding and secretion capability, and safety as host over E. coli. But S. cerevisiae has shortcomings of low expression level which is just 20% of that of E. coli. To solve this problem, directed evolution method was tried to enhance the GAP promoter strength of S. cerevisiae in this study. As result, modified GAP promoter that has increased expression level of about 360% compared to that of wild type was selected.

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Effect of Promoters on the Heme Production in a Recombinant Corynebacterium glutamicum (재조합 Corynebacterium glutamicum으로부터 헴첼 생산에 미치는 프로모터의 효과)

  • Yang, Hyungmo;Kim, Pil
    • Microbiology and Biotechnology Letters
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    • v.47 no.3
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    • pp.337-342
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    • 2019
  • We published that bacterial heme was over-produced in a recombinant Corynebacterium glutamicum expressing 5-aminolevulinic acid synthase ($hemA^+$) under control of a constitutive promoter ($P_{180}$) and the heme-producing C. glutamicum had commercial potentials; as an iron feed additive for swine and as a preservative for lactic acid bacteria. To enhance the heme production, the $hemA^+$ gene was expressed under controls of various promoters in the recombinant C. glutamicum. The $hemA^+$ expression by $P_{gapA}$ (a constitutive glycolytic promoter of glyceraldehyde-3-phosphate dehydrogenase) led 75% increase of heme production while the expression by $P_{H36}$ (a constitutive, very strong synthetic promoter) resulted in 50% decrease compared with the control ($hemA^+$ expression by $P_{180}$ constitutive promoter). The $hemA^+$ expression by a late log-phase activating $P_{sod}$ (an oxidative-stress responding promoter of superoxide dismutase) led 50% greater heme production than the control. The $hemA^+$ expression led by a heat-shock responding chaperone promoter ($P_{dnaK}$) resulted in 121% increase of heme production at the optimized heat-shock conditions. The promoter strength and induction phase are discussed based on the results for the heme production at an industrial scale.

Versatile Luciferase Reporter Plasmids for Transcription Studies in Diverse Eukaryotic Cells (다양한 진핵생물 세포에서 전사 연구에 사용될 수 있는 Luciferase Reporter Plasmid의 개발)

  • 조영석;한동욱;백금희;박승필;윤상순;임운기;김정락;김한도;강호성
    • The Korean Journal of Zoology
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    • v.39 no.4
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    • pp.378-386
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    • 1996
  • We have developed a couple of new luciferase reporter plasmida with very low background reporter actlvltlea. One can be used to measure the promoter strength, after insertion of some promoter fragment into the reporter plasmid, and the other, with very low basal promoter actlvltlea, allis In studying eukaryotic transcriptional regulators. The latter reporter plasmid contains such cli elements as a 17 nucleotide long inftlator, Spl.blndIng sftes, GAL4 binding sltea, and bInding sitea for a certain Drosophila homeodomain proteins. In an attempt to construct an improved reporter plaimid by fadlltating transcriptional termination and minimizing any interference by cryptic promoters which may be preaent in the reporter pleamld DNA, we have inserted transsrlptional termination-related signals, a three tandem repeat of SV4O polyadenylatlon signal (AAA) and the putative transcrtptional termination signal (UMS) of the mouse c-mos gene, Into just upstream of the initIator, and the promoter actlvitiea were measured by a transIent expression assay employing the Drosophila Schneider line 2 cells. As expected, the basal promoter activitIes decreased maximally when both transcription termination related elements were inserted. Moreover, the reporter plasmld with the two elements allowed more sensitive measurement of transcriptional activation than the reporter piasmid without them. Theae reporter plasmids can be used for studying transcriptional regulators of higher organisms Including mammals as well as Droiophlla melanogaiter.

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A Study on the Bond Strength of BCB-bonded Wafers (BCB 수지로 본딩한 웨이퍼의 본딩 결합력에 관한 연구)

  • Kwon, Yongchai;Seok, Jongwon;Lu, Jian-Qiang;Cale, Timothy;Gutmann, Ronald
    • Korean Chemical Engineering Research
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    • v.45 no.5
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    • pp.479-486
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    • 2007
  • Four point bending is used to study the dependences of bond strength of benzocyclobutene(BCB) bonded wafers and BCB thickness, the use of an adhesion promoter, and the materials being bonded. The bond strength depends linearly on BCB thickness, due to the thickness-dependent contribution of the plastic dissipation energy of the BCB and thickness independence of BCB yield strength. The bond strength increases by about a factor of two with an adhesion promoter for both $2.6{\mu}m$ and $0.4{\mu}m$ thick BCB, because of the formation of covalent bonds between adhesion promoter and the surface of the bonded materials. The bond strength at the interface between a silicon wafer with deposited oxide and BCB is about a factor of three higher than that at the interface between a glass wafer and BCB. This difference in bond strength is attributed to the difference in Si-O bond density at the interfaces. At the interfaces between plasma enhanced chemical vapor deposited (PECVD) oxide coated silicon wafers and BCB, and between thermally grown oxide on silicon wafers and BCB, 12~13 and $15{\sim}16bonds/nm^2$ need to be broken. This corresponds to the observed bond energies, $G_0s$, of 18 and $22J/m^2$, respectively. Maximum 7~8 Si-O $bonds/nm^2$ are needed to explain the $5J/m^2$ at the interfaces between glass wafers and BCB.

Association of RASSF1A Promoter Methylation with Lung Cancer Risk: a Meta-analysis

  • Huang, Ying-Ze;Wu, Wei;Wu, Kun;Xu, Xiao-Ning;Tang, Wen-Ru
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.23
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    • pp.10325-10328
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    • 2015
  • RASSF1A, regarded as a candidate tumor suppressor, is frequently silenced and inactivated by methylation of its promoter region in many human tumors. However, the association between RASSF1A promoter methylation and lung cancer risk remains unclear. To provide a more reliable estimate we conducted a meta-analysis of cohort studies to evaluate the potential role of RASSF1A promoter methylation in lung carcinogenesis. Relevant studies were identified by searches of PubMed, Web of Science, ProQest and Medline databases using the following key words: 'lung cancer or lung neoplasm or lung carcinoma', 'RASSF1A methylation' or 'RASSF1A hypermethylation'. According to the selection standard, 15 articles were identified and analysised by STATA 12.0 software. Combined odds ratio (OR) and 95% confidence interval (CI) were used to assess the strength of the association between RASSF1A promoter methylation and lung cancer risk. A chi-square-based Q test and sensitivity analyses were performed to test between-study heterogeneity and the contributions of single studies to the final results, respectively. Funnel plots were carried out to evaluate publication bias. Overall, a significant relationship between RASSF1A promoter methylation and lung cancer risk (OR, 16.12; 95%CI, 11.40-22.81; p<0.001) with no between-study heterogeneity. In subgroup analyses, increased risk of RASSF1A methylation in cases than controls was found for the NSCLC group (OR, 13.66, 95%CI, 9.529-19.57) and in the SCLC group (OR, 314.85, 95%CI, 48.93-2026.2).

Expression System for Optimal Production of Xylitol Dehydrogenase (XYL2) in Saccharomyces cerevisiae (출아효모에서 xylitol dehydrogenase (XYL2)의 최적 생산을 위한 발현 시스템 구축)

  • Jung, Hoe-Myung;Kim, Yeon-Hee
    • Journal of Life Science
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    • v.27 no.12
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    • pp.1403-1409
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    • 2017
  • In this study, the xylitol dehydrogenase (XYL2) gene was expressed in Saccharomyces cerevisiae as a host cell for ease of use in the degradation of lignocellulosic biomass (xylose). To select suitable expression systems for the S.XYL2 gene from S. cerevisiae and the P.XYL2 gene from Pichia stipitis, $pGMF{\alpha}-S.XYL2$, $pGMF{\alpha}-P.XYL2$, $pAMF{\alpha}-S.XYL2$ and $pAMF{\alpha}-P.XYL2$ plasmids with the GAL10 promoter and ADH1 promoter, respectively, were constructed. The mating factor ${\alpha}$ ($MF{\alpha}$) signal sequence was also connected to each promoter to allow secretion. Each plasmid was transformed into S. cerevisiae $SEY2102{\Delta}trp1$ strain and the xylitol dehydrogenase activity was investigated. The GAL10 promoter proved more suitable than the ADH1 promoter for expression of the XYL2 gene, and the xylitol dehydrogenase activity from P. stipitis was twice that from S. cerevisiae. The xylitol dehydrogenase showed $NAD^+$-dependent activity and about 77% of the recombinant xylitol dehydrogenase was secreted into the periplasmic space of the $SEY2102{\Delta}trp1/pGMF{\alpha}-P.XYL2$ strain. The xylitol dehydrogenase activity was increased by up to 41% when a glucose/xylose mixture was supplied as a carbon source, rather than glucose alone. The expression system and culture conditions optimized in this study resulted in large amounts of xylitol dehydrogenase using S. cerevisiae as the host strain, indicating the potential of this expression system for use in bioethanol production and industrial applications.