• Title/Summary/Keyword: expression vectors

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Hemocyte-specific Promoter for the Development of Transgenic Silkworm, Bombyx mori

  • Park, Seung-Won;Goo, Tae-Won;Kim, Seong-Ryul;Choi, Gwang-Ho
    • International Journal of Industrial Entomology and Biomaterials
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    • v.25 no.1
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    • pp.111-114
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    • 2012
  • In previous studies we have shown that a sw17255 gene was expressed in hemocyte-specific tissues of the silkworm, Bombyx mori L. (Lepidoptera: Bombycidae). It was verified that the sw17255 core promoter region contains elements that regulate the expression of this gene in hemocyte tissue; the selected promoter region spans nucleotides -1 to -2,112 upstream of the start codon. Each of the luciferase reporter gene expression vectors under the control of 4 different kinds of promoter candidates, (-2,112/-1), (-1,640/-1), (-1,169/-1) and (-579/-1), and the control reporter plasmid DNA, were introduced into B. mori larval coelom by direct injection using a syringe. The promoter candidate [E] (-579/-1) showed more than 1.67 fold transcriptional activity compared to control promoter activity. Higher productivity of an expressed gene in the transgenic silkworm by this promoter combination could be achieved in the near future. The foreign recombinant protein could be easily harvested from the blood of the transgenic silkworm.

Expression of Starch-degrading Genes in Escherichia Coli and Kactococcus Lactis

  • Jeong, Jong-Jin;Kim, Tea-Youn;Moon, Gi-Seong;Lee, Hyo-Jeong;Kim, Jong-Sang;Kim, Jeong -Hwan
    • Preventive Nutrition and Food Science
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    • v.3 no.1
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    • pp.98-104
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    • 1998
  • As an efffort ot construct LAB (latice acid bacteria), capable of utilizing starch as fermentation substrate without the aid of externally supplied enzymes, plasmid vectors containing the amyL($\alpha$-amylase/pullulansase gene) from Clostridium thermophydrosulfuricum, and glucoamylase cDNA from Asperigillus shirousamii were constructed and introduced itno E. coli and L. lactis. For expression in procaryotes , 1.9kb glucoamylase cDNA encoding the mature form of enzyme was PCR amplified and translationaly fused to a PCR amplified 260 bp fragment containing the promotor and secretion signals of amyl in the same reading frame. The production of $\alpha$-amylase, Apu, and glucoamlase in E. coli and L. lactis was confirmed by enzyme assay and zymography . Enzymeswere detected in both cellpellets and supernatants, indicating theworking of scretion signals in heterologous hosts. The efficiencies of secretion were varibale depending on the gene and host. The highest $\alpha$- amylase acitivity observed was 1.1 units and most activiity was detected from thecell pellets. The degree of gene expression in both hosts and the effect on the growth of hosts were examined.

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A Design and Implementation of 3D Facial Expressions Production System based on Muscle Model (근육 모델 기반 3D 얼굴 표정 생성 시스템 설계 및 구현)

  • Lee, Hyae-Jung;Joung, Suck-Tae
    • Journal of the Korea Institute of Information and Communication Engineering
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    • v.16 no.5
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    • pp.932-938
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    • 2012
  • Facial expression has its significance in mutual communication. It is the only means to express human's countless inner feelings better than the diverse languages human use. This paper suggests muscle model-based 3D facial expression generation system to produce easy and natural facial expressions. Based on Waters' muscle model, it adds and used necessary muscles to produce natural facial expressions. Also, among the complex elements to produce expressions, it focuses on core, feature elements of a face such as eyebrows, eyes, nose, mouth, and cheeks and uses facial muscles and muscle vectors to do the grouping of facial muscles connected anatomically. By simplifying and reconstructing AU, the basic nuit of facial expression changes, it generates easy and natural facial expressions.

Expression and Biochemical Characterization of CMCase Gene of Rhizobium fredii Usda193 in Escherichia coli (E. coli에서 근류균 섬유소 분해효소 유전자의 발현 및 생화학적 특성조사)

  • Yun, Ho-Jong;Park, Yong-Woo;Lim, Sun-Tech;Kang, Kyu-Young;Yun, Han-Dae
    • Microbiology and Biotechnology Letters
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    • v.23 no.3
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    • pp.275-281
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    • 1995
  • From the plasmid pYA300 carring a CMCase of Rhizobium fredii USDA193 plasmid was subcloned into pBluescript II KS(+)/pBluescript II SK(+) vectors and designated pYA500 and pYA600, respectively. Escherchia coli cells transformed with pYA500 porduced the CMCase more than with pYA600. The orientation of the cloned fragment in pBluescript vector had the effect on gene expression in E. coli background. When the 1.7 kb CMCase gene fragment of R. fredii USDA193 was hybridized to EcoRI-digested total DNA from R. meliloti and R. fredii USDA 191 the unique bands hybridized respectively, indicating that some genetic diversity exists in the EcoRI restriction enzyme site for CMCase gene in Rhizobium strains. The optimum pH of enzyme activity was 7 and the optimum temperature of that was nearly 37$\circ$C. The cellulase-minus derivatives of pYA500 were constructed by Tn5 insertional mutation. Among 6000 transconjugants, two mutant plasmids (designated pYA500::Tn5a and pYA500::Tn5b) were detected from the cellulase- negative transconjugants. The product of CMCase gene was analyzed by one dimensional SDS- PAGE of the cell extracts. About 45 kDa protein was considered to be a product of CMCase gene.

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Transgenesis in Fish: Indian Endeavour and Achievement

  • Pandian, T.J
    • Journal of Aquaculture
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    • v.16 no.1
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    • pp.51-58
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    • 2003
  • The first Indian transgenic fish was generated in 1991 using borrowed constructs from foreign sources. To construct transformation vectors for the indigenous fishes, growth hormone genes of rohu (r-CH), Labeo rohita and catfish, Heteropneustes fossilis were isolated, cloned and sequenced; their fidelity was confirmed in prokaryotic and eukaryotic systems. A vector was constructed with grass carp b-actin promoter driving the expression of r-GH. Rohu eggs are large. fragile and swell 2~3 times. when fertilized. Hence they were amenable only for electroporated sperm-mediated gene transfer. Accordingly, the sperm electroporation technique was standardized to ensure 25% hatchling survival and 37% Presumptive transgenics without suffering any deformity. Southern analysis confirmed genomic integration in 15% of the tested individuals (Ti) belonging to family lines 2 and 3: another 25% of the Juveniles (Te) were also proved transgenic but with the transgene persisting extrachromosomally for longer than 1 to 2 years. perhaps due to the presence of replicon in the vector. Transgenics belonging to different family lines grew 6~8 times faster than the respective controls. Difference in growth trends of Ti and Te within a family line was not significant. In the Ti family 3 remarkable growth acceleration was sustained for a period longer than 36 weeks but in those of family 2, it gradually decreased. All transgenic fishes including the rohu converted the food at a significantly higher efficiency. Barring the transgenic mudloach, all the other transgenic fishes consumed food at significantly reduced rate.

Expression of dirigent protein and Pinoresinol/Lariciresinol reductase genes of forsythia in transgenic potatoes.

  • Chuong, Tran-Van;Kim, Hyunsoon;Park, Jiyoung;Jaeyoul Joung;Jungwon Youm;Jaeheung Jeon
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2001.11a
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    • pp.49-58
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    • 2001
  • We tried to introduce two forsythia genes related in lignan biosynthesis, dirigent protein and pinoresinol/lariciresinol (P/L) reductase, into potatoes for accumulation of lignans in transgenic potatoes. We made binary vectors overexpressing dirigent protein gene and P/L reductase gene driven by a CaMV35S promoter and transformed into potatoes via Agrobacterium mediated transformation. And in order to control the metabolic flux of lignan biosynthesis pathway, we tried to inhibit chalcone synthase genes of potatoes by antisense inhibition technique also. We tried to use PCR screening method for selection of transgenic plants of different vectors. We tried to determine and compare lignan contents from different transgenic potato lines.

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Implementation of Underwater Simulation of a Net using OpenMP (OpenMP 병렬프로그램을 이용한 그물의 수중형상 시뮬레이션 구현)

  • Park, Myeong-Chul;Park, Seok-Gyu
    • Journal of the Korea Society of Computer and Information
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    • v.13 no.2
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    • pp.11-17
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    • 2008
  • The net shape effects by the various vectors in underwater. Each particle of the net calculating the effect of all vectors augments an accuracy and reality. But, the time complexity becomes larger because of huge calculation. The previous techniques reduced a physics reality. And embodied the underwater virtual reality which augments visual reality with simulation. In this paper, parallel processing the particles, it embodied the simulation which is satisfied a physical reality and time reality. The parallel processing used the OpenMP, and the reality graphic expression used the OpenGL. The simulation which this paper Proposes will be the possibility becoming the fundamental data for a model analysis or a specialist system from game and marine field.

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Expression of Dengue virus EIII domain-coding gene in maize as an edible vaccine candidate

  • Kim, Hyun A;Kwon, Suk Yoon;Yang, Moon Sik;Choi, Pil Son
    • Journal of Plant Biotechnology
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    • v.41 no.1
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    • pp.50-55
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    • 2014
  • Plant-based vaccines possess some advantages over other types of vaccine biotechnology such as safety, low cost of mass vaccination programs, and wider use of vaccines for medicine. This study was undertaken to develop the transgenic maize as edible vaccine candidates for humans. The immature embryos of HiII genotype were inoculated with A. tumefaciens strain C58C1 containing the binary vectors (V662 or V663). The vectors carrying nptII gene as selection marker and scEDIII (V662) or wCTB-scEDIII (V663) target gene, which code EIII proteins inhibite viral adsorption by cells. In total, 721 maize immature embryos were transformed and twenty-two putative transgenic plants were regenerated after 12 weeks selection regime. Of them, two- and six-plants were proved to be integrated with scEDIII and wCTB-scEDIII genes, respectively, by Southern blot analysis. However, only one plant (V662-29-3864) can express the gene of interest confirmed by Northern blot analysis. These results demonstrated that this plant could be used as a candidated source of the vaccine production.

A Study on the Length of DMZ and MDL (비무장지대 및 군사분계선의 길이에 관한 연구)

  • KIM, Chang-Hwan
    • Journal of the Korean Association of Geographic Information Studies
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    • v.22 no.1
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    • pp.19-27
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    • 2019
  • This study is to measure the length of the Demilitarized Zone and the Military Demarcation Line(MDL) on the Korean Peninsular. For this purpose, maps of the Armistice Agreement Volume II were used. These maps are nine sheets. In order to extract the MDL shown on the map, coordinates were assigned to the scanned image maps using the georeferencing module of ArcGIS based on the sheet line coordinates. The accuracy of the extracted vectors was checked by overlaying them on the maps of the Armistice Agreement Volume II. And I tried to validate these vectors through comparative analysis with vectors extracted from Kim(2007). Vectors extracted from Kim(2007) had errors in the curvilinear parts of the MDL, but the vectors extracted from this study exactly matched the MDL in the Armistice Agreement Volume II. The measured length is 239.42km(148.77miles). This means that the expression '155mile MDL' or '248km DMZ' in papers, reports or mass media has so far been inappropriate. I think this study will be able to provide information on the exact length of the DMZ in studies related with DMZ or in policy decisions by the national and local government. However, it is deemed necessary to verify this result by national organizations such as the NGII(National Geographic Information Institute). After these verification procedures, I hope that the national government will inform the people of the exact length of DMZ and MDL.

Construction of CpG Motif-enriched DNA Vaccine Plasmids for Enhanced Early Immune Response

  • Park Young Seoub;Hwang Seung Ha;Choi Cha-Yong
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.10 no.1
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    • pp.29-33
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    • 2005
  • A DNA vaccine methodology using eukaryote expression vectors to produce immunizing proteins in the vaccinated hosts is a novel approach to the development of vaccine and immuno-therapeutics, and it has achieved considerable success over several infectious diseases and various cancers. To further enhance its efficiency, attempts were made to develop novel plasmid vectors containing multiple immunostimulatory CpG motifs, for rapid and strong immune response. First, a 2.9 kb compact plasmid vector (pVAC), containing CMV promoter, polycloning site, BGH poly(A) terminator, ampicillin resistance gene and pBR322 origin was constructed. A pVAC-hEPO was also constructed, which contained a human erythropoietin gene, for evaluating the transfection efficiency of naked plasmid DNA both in vitro and in vivo. To examine the adjuvant effect of multi-CpG motifs on naked plasmid DNA, 22 and 44 enriched and unmethylated CpG motifs were introduced into pVAC to generate pVAC-ISS1 and pVAC-ISS2, respectively. $100{\mu}g$ of pSecTagB, pVAC, pVAC-ISS1 or pVAC-ISS2 were each injected intramuscularly into the tibilias anterior muscle of Balb/c mice. The level of interleukin-6 induced in the mice injected with pVAC-ISS1 and pVAC-ISS2 were significantly elevated after 12 hours, which were almost 2 and 2.5 times higher than that in the mice injected with pSecTagB, respectively. These results suggest that DNA vaccine plasmids with enriched CpG motifs can induce rapid secretion of interleukin-6 by lymphocytes. In conclusion, these vectors can contribute to the development of adjuvant-free DNA vaccinations against infectious diseases and various cancers.