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A novel chimeric promoter that is highly responsive to hypoxia and metals
Lee, J-Y,Lee, Y-S,Kim, J-M,Kim, K L,Lee, J-S,Jang, H-S,Shin, I-S,Suh, W,Jeon, E-S,Byun, J,Kim, D-K Nature Publishing Group 2006 Gene Therapy Vol.13 No.10
To develop a potent hypoxia-inducible promoter, we evaluated the usefulness of chimeric combinations of the (Egr-1)-binding site (EBS) from the Egr-1 gene, the metal-response element (MRE) from the metallothionein gene, and the hypoxia-response element (HRE) from the phosphoglycerate kinase 1 gene. In transient transfection assays, combining three copies of HRE (3 × HRE) with either EBS or MRE significantly increased hypoxia responsiveness. When a three-enhancer combination was tested, the EBS–MRE-3 × HRE (E–M–H) gave a hypoxia induction ratio of 69. The expression induced from E–M–H-pGL3 was 2.4-fold higher than that induced from H-pGL3 and even surpassed the expression from a human cytomegalovirus promoter-driven vector. The high inducibility of E–M–H was confirmed by validation studies in different cells and by expressing other cDNAs. Gel shift assays together with functional overexpression studies suggested that increased levels of hypoxia-inducible factor 1α, metal transcription factor-1 and Egr-1 may be associated with the high inducibility of the E–M–H chimeric promoter. E–M–H was also induced by hypoxia mimetics such as Co<SUP>2+</SUP> and deferoxamine (DFX) and by hydrogen peroxide. Gene expression from the E–M–H was reversible as shown by the reduced expression of the transgene upon removal of inducers such as hypoxia and DFX. In vivo evaluation of the E–M–H in ischemic muscle revealed that erythropoietin secretion and luciferase and LacZ expression were significantly higher in the E–M–H group than in a control or H group. With its high induction capacity and versatile means of modulation, this novel chimeric promoter should find wide application in the treatment of ischemic diseases and cancer.Gene Therapy (2006) 13, 857–868. doi:10.1038/sj.gt.3302728; published online 9 February 2006
김진석(J.S. Kim),김한수(H.S. Kim),이동윤(D.Y. Lee),이석우(S.W. Lee),강은구(E.G. Kang) 한국생산제조학회 2011 한국생산제조시스템학회 학술발표대회 논문집 Vol.2011 No.4
In recently, trends of the miniaturization of micro parts machining pattern by laser machining methods have been emerged. However, the conventional laser processing methods have limitations on the resolution. So, e-beam process has been considered as alternatives. Especially, e-beam lithography has been taken accounted of tools for micro and nano pattern. But e-beam lithography equipment have some disadvantages such as lower productivity and smaller processing area etc. than other micro parts machining tools. Therefore we have carried out the development of micro column electron beam lithography equipment for achieving the more higher productivity, lager processing area, and also higher resolution. And then we tried to develop e-beam lithography system with micro column type and using wafer lens for multi column e-beam lithography system and modular e-beam lithography unit. In this paper, we developed the electro static lens system, composed of accelerator lens, condenser lens, objective lens, beam blanker, stigmator and deflector.
소 품종별 Melanocortin Receptor 1(MC1R) 유전자의 유전자형 빈도에 관한 연구
한재용,김경남,오성종,정일정,김태헌,윤두학,박응우,이혜영,탁태영 한국동물자원과학회 2000 한국축산학회지 Vol.42 No.6
The melanocortin 1 receptor (MC1R) plays a central role in regulation of eumelanin(black/brown) and phaeomelanin (red / yellow) synthesis within the mammalian melanocyte and encoded by the classical Extension (E) coat color locus. The objectives of this study were carried out to observe the genotype frequencies of MC1R in 1,044 samples of imported beef and eight different cattle breeds including Hanwoo, Holstein, Angus, Brown-Swiss, Charolais, Limousin, Simmental and Hereford. The primers for the amplification of bovine MC1R gene were designed based on a bovine MC1R gene sequence (GenBank accession no. Y19103). A size of 350 by was amplified by polymerase chain reaction(PCR), digested with each or together of two different restriction enzyme, BsrFI and MspAII, and electrophoresed in 2.5% or 4% Metaphore agarose gel for determination of genotypes. One thousand and forty four samples including eight different cattle breeds and imported beef were determinated their genotypes by PCR-RFLP. Genotype frequencies of Hanwoo were 0.10 in E e and 0.90 in ee. Allele E^D was shown in all of Holstein and Angus breeds tested which have black coat color phenotypes. However, genotype frequencies of Limousin and Simmental breeds were 0.21 and 0.07 in E^DE^D, 0.00 and 0.03 in E^De^+ , 0.38 and 0.35 in E^De, respectively. These results of this study suggest that alleles of the bovine MC1R gene are associated strongly with coat color except for Simmental and Limousin breeds with dark brown, light yellow or mosaic coat color of red and brown. Therefore, it needs to be studied on relationship between A and E loci and coat color phenotypes. Furthermore, genotypes of the bovine MC1R gene can be used as a DNA marker for distinguishing. beef between Hanwoo and Holstein and Angus having black coat color phenotype.
Lee, K.H.,Choi, E.Y.,Hyun, M.S.,Jang, B.I.,Kim, T.N.,Kim, S.W.,Song, S.K.,Kim, J.H.,Kim, J.-R. S. Karger AG 2007 European surgical research Vol.39 No.4
<P><I>Background:</I> Proteolytic shedding of the ectodomain of a variety of transmembrane proteins, including cell-to-cell adhesion molecules, has been observed in solid cancers. We have investigated whether extracellular cleavage of E-cadherin mediated by matrix metalloproteinase-7 (MMP-7) is involved in hepatocyte growth factor (HGF) induced in vitro invasion in stomach cancer cells. <I>Methods:</I> The effects of HGF on the expression of E-cadherin/β-catenin and MMP-7 at both the protein and mRNA levels were assessed in stomach cancer cells, NUGC-3 and MKN-28, and in cells in which the expression of MMP-7 was downregulated by transfection with a MMP-7 short hairpin RNA plasmid. <I>Results:</I> Treatment with HGF increased the extracellular cleavage of E-cadherin and the release of MMP-7 and reduced the level of E-cadherin in a dose- and time-dependent manner. HGF treatment repressed the phosphorylation of β-catenin in a Triton-soluble fraction, but enhanced this phosphorylation in a Triton-insoluble fraction. The association of E-cadherin with β-catenin was decreased by HGF treatment in the Triton-soluble fraction. In addition, treatment of MMP-7 short hairpin RNA transfected NUGC-3 cells with HGF resulted in no extracellular cleavage of E-cadherin and also decreased the in vitro cell invasion. <I>Conclusions:</I> These results suggest that incubation with HGF mediated the release of MMP-7, resulting in extracellular cleavage of E-cadherin from stomach cancer cells. This might be a key mechanism in HGF-induced in vitro invasion and metastasis.</P><P>Copyright © 2007 S. Karger AG, Basel</P>
Balueva, E.N.,Lee, J.W. 한국응용곤충학회 2016 Journal of Asia-Pacific Entomology Vol.19 No.2
<P>In this paper, a taxonomic review of the genus Woldstedtius Carlson in Korea with a key to the twelve Eastern Palearctic species is provided. Two species-W. yokohamensis and W. karafutensis, which were described by Uchida from Japan-are re-described based on holotype re-examination and the study of Korean material. The male of W. karafutensis Uchida is reported for the first time. Three species, W corvinus sp. nov., W. nigrosignatus sp. nov., and W. pallidus sp. nov., are described as new. Five species, W. biguttatus Gravenhorst, W. citropectoralis Schmiedeknecht, W. holarcticus Diller, W. karafutensis Uchida, and W. yokohamensis Uchida, are reported from Korea for the first time. (C) 2015 Published by Elsevier B.V. on behalf of Korean Society of Applied Entomology, Taiwan Entomological Society and Malaysian Plant Protection Society.</P>
p34 is a novel regulator of the oncogenic behavior of NEDD4-1 and PTEN
Hong, S-W,Moon, J-H,Kim, J-S,Shin, J-S,Jung, K-A,Lee, W-K,Jeong, S-Y,Hwang, J J,Lee, S-J,Suh, Y-A,Kim, I,Nam, K-Y,Han, S,Kim, J E,Kim, K-p,Hong, Y S,Lee, J-L,Lee, W-J,Choi, E K,Lee, J S,Jin, D-H,Kim, Macmillan Publishers Limited 2014 CELL DEATH AND DIFFERENTIATION Vol.21 No.1
PTEN is one of the most frequently mutated or deleted tumor suppressors in human cancers. NEDD4-1 was recently identified as the E3 ubiquitin ligase for PTEN; however, a number of important questions remain regarding the role of ubiquitination in regulating PTEN function and the mechanisms by which PTEN ubiquitination is regulated. In the present study, we demonstrated that p34, which was identified as a binding partner of NEDD4-1, controls PTEN ubiquitination by regulating NEDD4-1 protein stability. p34 interacts with the WW1 domain of NEDD4-1, an interaction that enhances NEDD4-1 stability. Expression of p34 promotes PTEN poly-ubiquitination, leading to PTEN protein degradation, whereas p34 knockdown results in PTEN mono-ubiquitination. Notably, an inverse correlation between PTEN and p34/NEDD4-1 levels was confirmed in tumor samples from colon cancer patients. Thus, p34 acts as a key regulator of the oncogenic behavior of NEDD4-1 and PTEN.