• Title/Summary/Keyword: Heterologous

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Development of Non-protoplast transformation System in Aspergillus oryzae

  • Lee Jae Won;Hahm Young Tae
    • Proceedings of the Microbiological Society of Korea Conference
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    • 2000.10a
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    • pp.85-91
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    • 2000
  • Aspergillus oryzae is a filamentous fungus classified in the group Aspergillaceae Ascomycetes. It is an important microorganism for industrial production of enzymes and fermented food productions. It secrets large quantities of proteins or enzymes into the culture medium which makes this organism appealing for the production of heterologous proteins. Recently Electric field-mediated transformation method, electroporation, has been applied to fungal transformation. In this study, fungal transformation was carried out by bypassing the protoplast isolation step, decreasing the culturing time and non-protoplast transformation for the increment of transformation efficiency. Transformants were obtained with electroporation in optimal condition 2,500 voltage, 1,540 ohm and 0.50 capacitance. More than 1,000 transform ants were obtained with 6-10 hrs cultured mycelia without enzyme treatment, called non-protoplast transformation.

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Enhanced Flavonoid Production in Streptomyces venezuelae via Metabolic Engineering

  • Park, Sung-Ryeol;Ahn, Mi-Sun;Han, Ah-Reum;Park, Je-Won;Yoon, Yeo-Joon
    • Journal of Microbiology and Biotechnology
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    • v.21 no.11
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    • pp.1143-1146
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    • 2011
  • Metabolic engineering of plant-specific phenylpropanoid biosynthesis has attracted an increasing amount of attention recently, owing to the vast potential of flavonoids as nutraceuticals and pharmaceuticals. Recently, we have developed a recombinant Streptomyces venezuelae as a heterologous host for the production of flavonoids. In this study, we successfully improved flavonoid production by expressing two sets of genes predicted to be involved in malonate assimilation. The introduction of matB and matC encoding for malonyl-CoA synthetase and the putative dicarboxylate carrier protein, respectively, from Streptomyces coelicolor into the recombinant S. venezuelae strains expressing flavanone and flavone biosynthetic genes resulted in enhanced production of both flavonoids.

Application of Digital Polymerase Chain Reaction for Human Gene Detection in Heterologous Tissues (이종 조직에서 사람 유전자 검출을 위한 디지털 중합효소연쇄반응의 적용)

  • Kim, Jin-Hee
    • Proceedings of the Korea Contents Association Conference
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    • 2018.05a
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    • pp.561-562
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    • 2018
  • 디지털 중합효소연쇄반응(Digital PCR)은 3세대 PCR로 명명하며, 1세대인 일반 PCR과 2세대인 정량 PCR(Real-time PCR)의 단점을 보완하여 개발된 방법이다. Digital PCR System은 소량의 PCR 반응을 10만개 이상의 반응통(wall)에 적재하는 방식의 나노유체칩에서 쪼개어 증폭시킨 후, target DNA를 계수한다. Target DNA의 증폭 여부에 따라 positive(1)와 negative(0)로 digital signal처럼 받아들여 계수하고, 포아송 분포를 통해 target DNA의 copy를 계산해 최종적으로 샘플 microlitr당 Copy수로 결과 값을 확인할 수 있다. 본 연구에서는 종(種)이 다른 동물의 조직이 서로 섞여있을 때 사람의 조직을 탐색하는 방법으로 유전자 증폭을 할 경우, digital PCR의 유효성에 대해 증명하였다.

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Enhanced Gene Expression by Fusion to Rice-ubiquitin in Yeast

  • Kim, Young-Mi
    • Journal of Applied Biological Chemistry
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    • v.43 no.1
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    • pp.1-4
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    • 2000
  • Chloramphenicol acetyl CoA transferase (CAT) and angiotensin- converting enzyme inhibitory peptide (ACEI) were fused to C-terminal region of rice ubiquitin to examine the level of transcripts or enzyme activities in yeast. When two chimeric genes under an inducible Gall promoter control were transformed into Saccharomyces cerevisaie, both CAT and ACE inhibitory activities were enhanced by three to four-fold as compared to those containing no ubiquitin gene. However, the levels of transcripts of ubiquitin fused and un fused genes were not significantly different each other. Therefore, it was suggested that the expression of foreign genes was post-transcriptionally enhanced by fusion of plant ubiquitin in heterologous organisms such as yeast.

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A Study on the Partial Discharge Resistance Characteristic for Optimizing the Mixing Ratio of Heterogeneous Inorganic Insulated Materials for Environmentally Friendly GIS Spacer (친환경 GIS Spacer용, 이종 무기물 절연소재의 혼합비 최적화를 위한 부분방전 저항성 특성 연구)

  • Park, Jae-Jun
    • The Transactions of The Korean Institute of Electrical Engineers
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    • v.67 no.9
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    • pp.1189-1196
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    • 2018
  • 7 type composites (40, 45, 50, 55, 60, 65, and 70 wt.%)were prepared for the environmentally friendly GIS Spacer. Five kinds of samples were prepared for optimization of the filler content ratio (MS: MA = 1: 9, 3: 7, 5: 5, 7: 3, 9: 1) of epoxy / microsilica and microalumina. As a result of evaluation of the partial discharge resistance characteristic, surface erosion is generally slowed down as the fill amount of micro silica is increased. Also, partial discharge resistance characteristics for the development of insulating materials with optimal mixing ratios of heterologous showed a higher partial resistance of discharge and a decrease in erosion, as the filler content ratio of micro silica was larger. In the future, various researches such as electrical, mechanical, and thermal studies will be needed to develop insulating materials that can commercialize power devices in environmentally friendly insulating gas.

A STUDY ON ANTIGENICITY OF RECOMBINANT HUMAN GROWTH HORMONE (LBD-007) IN MICE AND GUINEA PIGS

  • Park, Jong-Il;Han, Sang-Seop;Roh, Jung-Koo
    • Toxicological Research
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    • v.9 no.1
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    • pp.125-132
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    • 1993
  • Antigenic potential of recombinant human growth hormone (LBD-007), a newly developed drug for growth hormone deficiency, was investigated in mice and guinea pigs. 1. Mice showed production of antibodies against LBD-009 (1.5IU/kg) with aluminum hydroxide gel(alum) as an adjuvant, Judaged by the heterologous anaphylaxis (PCA) test using rats. On the other hand, antibodies against ovalbumin (OVA) inoculated with alum were definitely detected. 2. In the studies with guinea pigs, both the inoculation of LBD-009 (0.15IU/kg-1.5IU/kg) only and of LBD-009 with complete Freund's adjuvant (CFA) as an adjuvant did produce weak positive reactions in homologous passive cutaneous anaphylaxis (PCA). On the other hand, the inoculation of ovalbumin with complete Freund's adjuvant (CFA) produced positive reaction in PCA.

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Studies on Hypersensitivity of Recombinant Hepatitis B Vaccine (LBD-008) in Mice and Guinea pigs

  • Park, Jong-Il;Ha, Chang-Su;Han, Sang-Seop
    • Biomolecules & Therapeutics
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    • v.2 no.2
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    • pp.108-113
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    • 1994
  • Toxicity study of recombinant hepatitis B vaccine (LBD-008), a newly developed drug for acute and chronic hepatitis, was investigated in mice and guinea pigs. 1. Mice showed no production of antibodies against LBD-008 inoculated with aluminum hydroxide gel (Alum) as an adjuvant, judged by the heterologous anaphylaxis (PCA) test using rats. On the other hand, antibodies against ovalbumin (OVA) inoculated with alum were definitely detected. 2. In the studies with guinea pigs, both the inoculation of LBD-008 only and of LBD-008 with complete Freund's adjuvant (CFA) as an adjuvant did not produce positive reactions in any of homologous active systemic anaphylaxis (ASA). On the other hand, the inoculation of ovalbumin with complete Freund's adjuvant (CFA) produced positive reaction in both of PCA and ASA. 3. These findings suggested that LBD-008 has no antigenic potential in mice or guinea pigs.

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Identification of a Cryptic Type III Polyketide Synthase (1,3,6,8-Tetrahydroxynaphthalene Synthase) from Streptomyces peucetius ATCC 27952

  • Ghimire, Gopal Prasad;Oh, Tae-Jin;Liou, Kwangkyoung;Sohng, Jae Kyung
    • Molecules and Cells
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    • v.26 no.4
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    • pp.362-367
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    • 2008
  • We identified a 1,134-bp putative type III polyketide synthase from the sequence analysis of Streptomyces peucetius ATCC 27952, named Sp-RppA, which is characterized as 1,3,6,8-tetrahydroxynaphthalene synthase and shares 33% identity with SCO1206 from S. coelicolor A3(2) and 32% identity with RppA from S. griseus. The 1,3,6,8-tetrahydroxynaphthalene synthase is known to catalyze the sequential decarboxylative condensation, intramolecular cyclization, and aromatization of an oligoketide derived from five units of malonyl-CoA to give 1,3,6,8-tetrahydroxynaphthalene, which spontaneously oxidizes to form 2,5,7-trihydroxy-1,4-naphthoquinone (flaviolin). In this study, we report the in vivo expression and in vitro synthesis of flaviolin from purified gene product (Sp-RppA).

Bacillus cellulyticus K-12 Crystalline Cellulose-Degrading Avicelase Gene and Expression in Eschterichia coli

  • Cheorl-Ho Kim;Woo
    • The Korean Journal of Food And Nutrition
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    • v.6 no.4
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    • pp.314-321
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    • 1993
  • We have cloned the Bacillus cellulyticus K-12 avicelase (Avi, E.C.3.2.1.4) gene (ace A) In E. coli. This was accompanied by using the vector PT7T3U 19 and Hind W -Hind m libraries of Bacillus cellulyticus K-12 chromosomal inserts created in 5.cofi. The Libraries were screened for the expression of avicelase by monitoring the immunoreaction of the anti-avicelase (immunoscreening). Positive clones (Ac-3, Ac-5, and Ac-7) contained the identical 3.5kb Hind III fragment as determined by restriction mapping and Southern hybridization, and expressed avicelase efficiently and constituvely using its own promoter in the heterologous host. From the immunoblotting analysis, a polypeptide which showed a CMCase activity with an Mr of 54000 was detected.

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