• 제목/요약/키워드: Cell transformation

검색결과 627건 처리시간 0.03초

진핵생물 유전자 조작을 위한 효모 vector계 이용에 관한 기초연구 -생효모 형질전환 최적조건과 숙주별 plasmid안정성에 관하여 - (Development of Yeast-Vector System for Eukaryotic Gene Cloning - Optimum Condition for Intact Yeast Cell Transformation and Plasmid Stability in the Transformants -)

  • 기우경;조성환;김범규;조무제
    • 한국미생물·생명공학회지
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    • 제14권2호
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    • pp.125-131
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    • 1986
  • 각 효모 숙주 및 vector에 따라 lithium염 처리에 의한 생효모형질전환 최적 조건을 얻기 위하여 5종의 효모 숙주(S. cerevisiae Dl3-lA, DKD-5D, DBY-746, MC-16 및 S2022D)에 3종의 효모plasmid vector(YRp 7, YEp 13 및 YIp 5 )의 형질전환 실험과 아울러 이들 각 형질전환체내에서 도입된 plasmid들의 안정성을 조사하였다. lithium 염의 경우 LiCl가 Li-acetate에 비하여 좋은 효과를 보였으며 LiCl처리에 의한 최적 형질전환 조건은 각 숙주-vector계에 있어서 공히 균체 배양 시간은 16시간 (5.4 $\times$ $10^6$ - 2.4$\times$$10^8$cells/$m{\ell}$ ) 내외, LiCl의 농도는 0.1-0.2M, PEG(4000)의 농도는 35%, induction시간은 60분 내외 열처리는 42$^{\circ}C$에서 5분간, LiCi처리 buffer는 0.1M tris-HCl(pH 7.6)에서 가장 높은 형질전환 빈도를 보였다. 한편 Protoplast 형질전환법과 형질전환빈도를 비교해 본 결과 DKD-5 D(YEp13)과 Dl3-lA(YRp7)의 경우는 protoplast법이 DKD-5D(YRp 7), 및 DBY-746(YEp 13)에서는 LiCl처리법이 형질전환 빈도가 높았으며 MC-16(YEp 13)의 경우는 양방법에서 공히 비교적 낮은 빈도를 보였다. 각 형질전환체내에서의 도입된 plasmid의 안정성은 선택배지에서 배양했을 경우- YRp 7이 Dl3-lA 및 DKD- 5D 숙주에서 70세대후에는 80-85%가 유실되었으나 YEp13은 DKD-5D 및 DBY-746에서 35%밖에 유실되지 않았으며 MC-16숙주에서는 55% 유실로서 비교적 안정하였다. 또한 비선택배지에 배양시에는 선택배지에 배양했을 경우 보다 안정성이 다소 낮았으나 같은 경향은 보였다.

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Characterization of the Cell-Surface Barriers to Plasmid Transformation in Corynebacterium glutamicum

  • Jang, Ki-Hyo;Paul J. Chambers;Chun, Uck-Han;Margare L.Britz
    • Journal of Microbiology and Biotechnology
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    • 제11권2호
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    • pp.294-301
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    • 2001
  • The effects of including glycine and isonicotinic acid hydrazide (INH) in the growth medium (Luria broth, LBG) on the subsequent lysozyme-imduced protoplast formation and transformation efficiency of Corynebacterium glutamicum were studied. The transformation efficiency of C. glutamicum AS019 increased up to 100-fold as the ocncentrationof glycine in the media increased from 0% to 5% (w/v), relative to cells grown in the absence of glycine. The presence of 5 mg/ml INH in the growth medium led to a further 10-fold increase in transformation efficiency. In addition, this transformation protocol was successfully applied to other strains of C. glutamicum. Both chemicals affected the mycolic acid attachment to the cell surface of C. glutamicum, when INH, the relative percentage of fatty acids of AS019 to the total lipids (mycolic acid plus fatty acids) decreased from 76.9% (in LBG) to 72.9% (in LBG-2% glycine) and 66.4% (in LBG-8 mg InG/ml), thereby suggeting that these chemicals also inhibit fatty acid synthesis.

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PREVENTION OF CANCER BY DIETARY FACTORS: TARGETING MAP KINASE/AP-1 SIGNAL TRANSDUCTION PATHWAYS

  • Dong, Zigang
    • 한국독성학회:학술대회논문집
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    • 한국독성학회 2001년도 International Symposium on Dietary and Medicinal Antimutgens and Anticarcinogens
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    • pp.29-30
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    • 2001
  • MAP kinase/ AP-1 signal transduction components are rapidly initiated by many extracellular stimuli, especially environmental carcinogens. We have investigated the role of MAP kinases (Erks, JNKs, and p38 kinases) and AP-1 signal transduction pathways in the process of cell transformation and carcinogenesis. Incubation of Cl 41 cells with tumor promoters such as TPA, EGF, arsenic, or TNF-$\alpha$ led to cell transformation and activation of MAP kinases.(omitted)

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Overcoming of Barriers to Transformation in Monocot Plants

  • Toyama Koichi;Bae, Chang-Hyu;Seo, Mi-Suk;Song, In-Ja;Lim, Yong-Pyo;Song, Pill-Soon;Lee, Hyo-Yeon
    • Journal of Plant Biotechnology
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    • 제4권4호
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    • pp.135-141
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    • 2002
  • Agrobacterium-mediated transformation has been unsuccessful for monocot plants except for a few important crops such as barley, rice, maize and wheat. We discussed here that a successful transformation of monocots demands certain critical conditions. The requirements for an efficient transformation are a selection of target tissues competent for plant regeneration and Agrobacterium-infection, and various factors promoting Agrobacterium-infection. The factors were divided into two to activate Agrobacterium and to increase plant cell's susceptibility against Agrobacterium. Optimization of these factors significantly increased transformation efficiency of zoysia grass and rice plants. A technical improvement in transformation system for monocots will promote improvement of the breed as well as a study of gene functions in monocots.

액정셀의 전기광학적 편광투과특성을 이용한 액정표시소자의 최적설계: I. 단색광의 경우 (Optimal Design of Liquid Crystal Display Devices Using Electro-optic Characteristics of Liquid Crystal Cell: I. Monochromatic Case)

  • 노봉규;김규석;김진승
    • 한국광학회지
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    • 제5권3호
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    • pp.411-417
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    • 1994
  • 액정층을 지나는 빛의 편광상태의 변화를 뽀앙카레공에서의 회전변환으로 나타내어, 액정층에 걸어준 전압에 대한 회전변환이 회전각과 회전축의 변화를 쟀다. 이로부터 액정셀에 걸린 전압이 비선택전압에서 선택전압으로 바뀔 때의 회전변환의 회전각이 $180^{\circ}$에 가장 가까운 구동전압을 결정하고, 이 전압으로 액정셀을 구동할 때 화면의 콘트라스트 및 투과율이 최대가 되는 현광판과 검광판의 투과축의 방향을 회전변환의 일반적인 특성을 써서 결정하고 실험적으로 확인하였다.

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Protective effect and mechanism of ginsenoside Rg2 on atherosclerosis

  • Qianqian Xue;Tao Yu;Zhibin Wang;Xiuxiu Fu;Xiaoxin Li;Lu Zou;Min Li;Jae Youl Cho;Yanyan Yang
    • Journal of Ginseng Research
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    • 제47권2호
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    • pp.237-245
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    • 2023
  • Background: Ginsenoside Rg2 (Rg2) has a variety of pharmacological activities and provides benefits during inflammation, cancer, and other diseases. However, there are no reports about the relationship between Rg2 and atherosclerosis. Methods: We used 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) to detect the cell viability of Rg2 in vascular smooth muscle cells (VSMCs) and human umbilical vein endothelial cells (HUVECs). The expression of inflammatory factors in HUVECs and the expression of phenotypic transformation-related marker in VSMCs were detected at mRNA levels. Western blot method was used to detect the expression of inflammation pathways and the expression of phenotypic transformation at the protein levels. The rat carotid balloon injury model was performed to explore the effect of Rg2 on inflammation and phenotypic transformation in vivo. Results: Rg2 decreased the expression of inflammatory factors induced by lipopolysaccharide in HUVECs-without affecting cell viability. These events depend on the blocking regulation of NF-κB and p-ERK signaling pathway. In VSMCs, Rg2 can inhibit the proliferation, migration, and phenotypic transformation of VSMCs induced by platelet derived growth factor-BB (PDGF-BB)-which may contribute to its anti-atherosclerotic role. In rats with carotid balloon injury, Rg2 can reduce intimal proliferation after injury, regulate the inflammatory pathway to reduce inflammatory response, and also suppress the phenotypic transformation of VSMCs. Conclusion: These results suggest that Rg2 can exert its anti-atherosclerotic effect at the cellular level and animal level, which provides a more sufficient basis for ginseng as a functional dietary regulator.

우췌성 백반증에서 기원한 우췌성 암종의 치험례 (A CASE PRESENTATION OF VERRUCOUS CARCINOMA ORIGINATED FROM VERRRUCOUS LEKOPLAKIA)

  • 최문기
    • Maxillofacial Plastic and Reconstructive Surgery
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    • 제28권4호
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    • pp.339-347
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    • 2006
  • Oral leukoplakia is the most common premalignant lesion and malignant transformation has been reported from verrucal lekoplakia. Homogenous, benign leukoplakia develops into a line of squamous cell carcinoma such as verrrucous carcinoma, papillary squamous cell carcinoma and invasive squamous cell carcinoma. Early diagnosis and intervention of premalignant leukoplakia is up-most important to prevent transformation into a oral squamous cell carcinoma. Any change in surface, size and color warrants repeated biopsy. If verrucous carcinoma is evidently derived from the previous leukoplakia, wide surgical excision and periodic follow up is needed. Surgically removed lesion of leukoplakia has the tendency to recur. Follow-up is very important to patient and clinician. Although many therapies have been reported to oral leukoplakia and verrucal carcinoma, accepted treatment principle is not exist so far. But surgical removal is recommended as the treatment of choice.

액상 투과전자현미경 분석기법 소개 및 최신 연구동향 (Current Status of Liquid-cell Transmission Electron Microscopy)

  • 홍재영;천동원
    • 세라미스트
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    • 제22권4호
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    • pp.417-428
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    • 2019
  • Even though, nanoscale materials of various shapes and compositions have been synthesized in the liquid, their underlying growth and transformation mechanisms are not well understood due to a lack of analytical methods. The advent of liquid cell for transmission electron microscope (TEM) enables the direct imaging of chemical reactions that occur in liquids with nanometer resolution of the electron microscope (EM). Here, the technical development of liquid cell TEM equipment and their applications to the study of nanomaterials analysis in liquid are discussed. Also new findings discovered through liquid cell TEM studies such as nucleation & growth, coalescence process and transformation are discussed.

두경부에 발생한 Progressive Transformation of Germinal Centers(PTGC) 환자 2예 (Two Cases of Progressive Transformation of Germinal Centers(PTGC) in Head and Neck Area)

  • 한원규;김태민;윤희철;염건휘;오경호;권순영
    • 대한두경부종양학회지
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    • 제31권1호
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    • pp.34-38
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    • 2015
  • Progressive transformation of germinal centers(PTGC) is a rare benign lymph node enlargement characterized by chronic lymphadenopathy and the presence of germinal centers with fused and expanded mantles within a lymph node that are 3-5 times the size of a typical reactive follicle. Although PTGC is reported in 3.5-10% of chronic lymphadenopathy, head and neck surgeons do not often experience this condition and its clinical implications. Althoug PTGC is benign disease, it could not be excluded the association with other malignant lymphoid diseases. Therefore, periodic follow up of PTGC patient could be required. We report two cases at this study ; a 16 years old man who visited out-patient clinic due to palpable mass of right cervical area, several months ago, and a 35 years old man who visited out-patient clinic for evaluating right cervical mass. Both of two patients were diagnosed with PTGC. In this study, we focus on the clinic-pathologic features of this uncommon disease along with other literature reviews.

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전기천공법에 의한 Bacillus brevis P176-2의 형질전환 (Transformation of Bacillus brevis P176-2 with Plasmid DNA by Electroporation)

  • 채기수;엄경일
    • 한국식품영양학회지
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    • 제5권2호
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    • pp.77-83
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    • 1992
  • The optimum conditions and mechanisms for the plasmid-mediated genetic transformation of intact cells of Bacillus brevis Pl76-2, an extracellular protein producing bacterium by electroporation were investigated. It was found that pUB110 Plasmid DNA can be introduced into intact bacterial cells by electroporation. The frequency of transformation by this electroporation system depended upon the initial electric field strength, the capacity of the electric discharge capacitor, growth stage, number of successive pulses and composition of electroporation buffer. It was effective for transformation that cells were harvested, washed and resuspended with HSM [7M HEPES(PH 7.4), 272mM sucrose, 1 mM MgCl2] electroporation buffer when cell growth was attained to 1.2 at OD660. A maximum frequency of transformation of 2.40$\times$104 transformants per$\mu$g plasmid DNA was obtained by two succesive Pulses with an initial electric field strength of 12.5kV/cm and with a capacitance of 7.3uF.

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