• 제목/요약/키워드: expression construct

검색결과 399건 처리시간 0.023초

Host-Induced gene silencing of fungal pathogenic genes confer resistance to fungal pathogen, Magnaporthe Oryzae in rice

  • Jin, Byung Jun;Chun, Hyun Jin;Kim, Min Chul
    • 한국작물학회:학술대회논문집
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    • 한국작물학회 2017년도 9th Asian Crop Science Association conference
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    • pp.134-134
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    • 2017
  • Recently, host-induced gene silencing (HIGS) system has been successfully applied into development of resistant crops against insects, fungal and viral pathogens. To test HIGS-mediated resistance in rice against rice blast fungus, Magnaporthe oryzae, we first tested possibility of movement of small non-coding RNA from rice cells to rice blast fungus. The rice blast fungus expressing GFP transgene were inoculated to transgenic rice plants ectopically expressing dsRNAi construct targeting fungal GFP gene. Expression of dsRNAi construct for GFP gene in transgenic plants significantly suppressed GFP expression in infected fungal cells indicating that small RNAs generated in plant cells can move into infected fungal cells and efficiently suppress the expression of fungal GFP gene. Consistent with these results, expression of dsRNAi constructs against 3 fungal pathogenic genes of M. oryzae in transgenic rice specifically and efficiently suppressed not only the expression of fungal pathogenic genes, but also fungal infection. The conidia of M. oryzae applied on leaf sheath of transgenic rice expressing dsRNAs against 3 fungal pathogenic genes showed abnormal development of primary hyphae and malfunction of appressorium, which is consistent with the phenotypes of corresponding fungal knock-out mutants. Taken these results together, here, we suggest a novel strategy for development of antifungal crops by means of HIGS system.

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진화연산에 기반한 유전자 발현 데이터로부터의 유전자 상호작용 네트워크 구성 (Construction of Gene Interaction Networks from Gene Expression Data Based on Evolutionary Computation)

  • 정성훈;조광현
    • 제어로봇시스템학회논문지
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    • 제10권12호
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    • pp.1189-1195
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    • 2004
  • This paper investigates construction of gene (interaction) networks from gene expression time-series data based on evolutionary computation. To illustrate the proposed approach in a comprehensive way, we first assume an artificial gene network and then compare it with the reconstructed network from the gene expression time-series data generated by the artificial network. Next, we employ real gene expression time-series data (Spellman's yeast data) to construct a gene network by applying the proposed approach. From these experiments, we find that the proposed approach can be used as a useful tool for discovering the structure of a gene network as well as the corresponding relations among genes. The constructed gene network can further provide biologists with information to generate/test new hypotheses and ultimately to unravel the gene functions.

Maltose Binding Protein 융합단백질에 의한 인간유래의 앤지오스타틴과 앤도스타틴의 대장균에서 수용성 단백질발현 (Soluble Expression of Human Angiostatin and Endostatin by Maltose Binding Protein (MBP) Fusion in E. coli)

  • 박선열;최신건
    • 산업기술연구
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    • 제28권B호
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    • pp.59-63
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    • 2008
  • Rapid production of therapeutic proteins such as angiostatin and endostatin angiogenic inhibititors has been highly demanded for cancer treatment. In this regard, recombinant human angiostatin and endostatin were successfully expressed as soluble forms by maltose binding protein (MBP)-mediated fusion expression in Escherichia coli. PCR amplified, angiostatin and endostatin genes from human placenta cDNA library were inserted into an expression vector pMAL-c2e to construct prokaryotic expression vectors, pMAL-c2e/AS and pMAL-c2e/ES, respectively. Recombinant angiostatin and endostatin were efficiently expressed in E. coli origami (DE3) after IPTG induction and protein expression were confirmed by SDS-PAGE analyses. The expressed recombinant proteins were purified near homogenity using an amylose affinty column chromatography. In contrast that previous E. coli expressions were all insoluble, our results first time demonstrated that MBP fused human angiostatin and endostatin were soluble in E. coli.

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Effects of the Vitreoscilla Hemoglobin Gene on the Expression of the Ferritin Gene in Escherichia coli

  • Chung, Yun-Jo;Kim, Kyung-Suk;Jeon, Eun-Soon;Park, Kie-In;Park, Chung-Ung
    • BMB Reports
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    • 제31권5호
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    • pp.503-507
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    • 1998
  • To investigate the effects of the Vitreoscilla hemoglobin (VHb) gene on the production of a heterologous protein, a comparative expression system for VHb and ferritin was constructed. First, the VHb gene was inserted into the downstream and upstream regions of the ferritin gene to construct pHF2 and pHF3, respectively. Next, the two plasmids pACHB1 and pVUTFH10, having the VHb gene and the ferritin gene respectively, were constructed in order to express the two genes in different plasmids by using a coplasmid expression system. It was observed that the cell growth was improved in all strains containing the VHb gene. Furthermore, in our coplasmid expression system, the presence of the VHb gene increased production of the ferritin by 1.8 times, as much as that in a strain not having the VHb gene.

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An approach to modeling a bionic self organization and functional expression

  • Yokoi, Hiroshi;Kakazu, Yukinori
    • 제어로봇시스템학회:학술대회논문집
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    • 제어로봇시스템학회 1992년도 한국자동제어학술회의논문집(국제학술편); KOEX, Seoul; 19-21 Oct. 1992
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    • pp.334-339
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    • 1992
  • This study is concerned how to construct a model of life as physical/ mathematical representation. This model is called here a bionic model and vibrating potential field is introduced as fundamental world background of the model. Namely, required information creating/ processing/ controlling are done on this field. Especially this paper reports how to realize the simulation of a bionic self organization and its functional expression based on the mutual actions among a set of life units.

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Expression of de novo Designed High Nutritional Peptide (HEAAE) in Tobacco

  • Kim, Jae-Ho;Lee, Chang-Kook;Hong, Bun-Shik
    • Journal of Microbiology and Biotechnology
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    • 제7권2호
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    • pp.138-143
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    • 1997
  • We have designed and constructed a gene encoding novel high essential amino acid encoding protein(HEAAE). The resultant DNA fragment was tested for in vitro and in vivo expression and then cloned into plant expression vector pBI121, under the control of the cauliflower mosaic virus 35S promoter. Agrobacterium tumefaciens, strain LBA4404, was subsequently transformed with this new construct and Nicotiana tabacum var. Xanthi transgenic plants were obtained. DNA analysis by Southern procedure confirmed the presence of the multi-copy number of genes in the transformed plants. Analysis of RNA and protein synthesized in these transgenic plants demonstrated the stable expression of this gene.

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LL과 LR에서의 효율적인 수식 타입 체킹에 대한 연구 (A Research on the Efficient Type-Checking for an Expression Using LL and LR)

  • 정용주
    • 한국인터넷방송통신학회논문지
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    • 제11권1호
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    • pp.177-183
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    • 2011
  • 수식의 자료형 확인을 위하여 많이 사용하는 방법의 하나가 속성문법이다, 그런데 이 속성문법은 구성하기가 어려운 단점이 있다. 파싱 과정을 잘 이해하고 속성문법의 두 개의 속성들을 사용하여야 하기 때문이다. 그래서 본 논문에서는 이 수식 속성문법의 구성을 보다 쉽게 하기 위한 방법을 제시한다. 문법 구성을 통하여 하나의 속성만으로 수식의 자료형 확인이 가능함을 보여준다.

Introduction and Expression of Foreign Genes in Rice Cells by Particle Bombardment

  • Jeon, Jong-Seong;Jung, Hou-Sung;Sung, Soon-Kee;Lee, Jong-Seob;Choi, Yang-Do;Kim, Han-Jip;Lee, Kwang-Woong
    • Journal of Plant Biology
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    • 제37권1호
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    • pp.27-36
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    • 1994
  • For establishing a transformation system of rice, an efficient introduction of foreign genes into embryogenic cell suspension by particle bombardment was conducted. The particle inflow gun based on the acceleration of DNA-coated tungsten particles using pressurized helium was constructed for delivery of DNA into rice cells. Several bombardment parameters were optimized using the transient expression of GUS gene. The conditions that gave the highest GUS gene expression of about 1000 blue spots per g fresh weight of bombarded cells include treatment of the cells with 0.5 M osmotic pressure, and use of the 410 kPa helium, 110 mm target distance, 13 mm syringe filter holder and 5 $\mu$L DNA/tungsten mixtures. It was also confirmed that rice actin promoter-intron construct gave the highest expression of all promoter-sequences studied. Eight weeks after the bombardment, stably transformed calluses were obtained on the selection medium containing 100 mg/L G418 and showed the strong activity in in situ GUS assay.

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Identification of the Regulatory Region Responsible for Vascular Tissue-Specific Expression in the Rice Hd3a Promoter

  • Pasriga, Richa;Cho, Lae-Hyeon;Yoon, Jinmi;An, Gynheung
    • Molecules and Cells
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    • 제41권4호
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    • pp.342-350
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    • 2018
  • Flowering time is determined by florigens. These genes include, Heading date 3a (Hd3a) and Rice FT 1 (RFT1) in rice, which are specifically expressed in the vascular tissues of leaves at the floral transition stage. To study the cis-regulatory elements present in the promoter region of Hd3a, we generated transgenic plants carrying the 1.75-kb promoter fragment of Hd3a that was fused to the ${\beta}$-glucuronidase (GUS) reporter gene. Plants expressing this construct conferred a vascular cell-specific expression pattern for the reporter gene. However, GUS was expressed in leaves at all developmental stages, including the early seedling stage when Hd3a was not detected. Furthermore, the reporter was expressed in roots at all stages. This suggests that the 1.75-kb region lackings cis-elements that regulate leaf-specific expression at the appropriate developmental stages. Deletion analyses of the promoter region indicated that regulatory elements determining vascular cell-specific expression are present in the 200-bp region between -245 bp and -45 bp from the transcription initiation site. By transforming the Hd3a-GUS construct to rice cultivar 'Taichung 65' which is defective in Ehd1, we observed that Ehd1 is the major regulatory element that controls Hd3a promoter activity.

Cooperation between Human DAF and CD59 in Protecting Cells from Human Complement-mediated Lysis

  • Xu, Li;Wu, Wenlan;Zhao, Zhouzhou;Shao, Huanjie;Liu, Wanhong;Liu, Hui;Li, Wenxin
    • BMB Reports
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    • 제39권6호
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    • pp.743-748
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    • 2006
  • The complement (C) regulatory proteins decay accelerating factor (DAF, CD55) and CD59 could protect host cells using different mechanisms from C-mediated damage at two distinct levels within the C pathway. Co-expression of DAF and CD59 would be an effective strategy to help overcome host C-induced xenograft hyperacute rejection. In this study, we made a construct of recombinant expression vector containing DAF and CD59 cDNA and the stable cell lines were obtained by G418 selection. Extraneous genes integration and co-expression were identified by PCR, RT-PCR and Western blot analysis. Human c-mediated cytolysis assays showed that NIH/3T3 cells transfected stably with pcDNA3-CD59, pcDNA3-DAF, and pcDNA3-CD59DAF-DP were protected from C-mediated damage and that synchronously expressed human CD59 and DAF provided the most excellent protection for host cells as compared with either human CD59 or DAF expressed alone. Therefore, the construct represents an effective and efficacy strategy to overcome C-mediated damage in cells and, ultimately, in animals.