• Title/Summary/Keyword: Cell Screening

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Establishment of a New Herbicides Screening Method Using Photoautotrophic Cultured Cell I. Studies on Culture Conditions for Developing Photoautotrophic Cells in Tobacco (광독립영양세포(光獨立營養細胞)를 이용(利用)한 새로운 제초제(除草劑) 선발법(選拔法) 확립(確立) I. 담배의 광독립영양세포(光獨立營養細胞) 육성(育成)을 위한 배양조건(培養條件)의 구명(究明))

  • Kim, K.U.;Suh, S.K.;Kwon, S.T.
    • Korean Journal of Weed Science
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    • v.11 no.1
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    • pp.68-73
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    • 1991
  • This study was conducted to determine the appropriate culture conditions for developing photoautotrophic cells from tobacco using plant growth regulators, sucrose and light condition in LS medium. The greatest callus induction was observed in the LS medium supplemented with $10^{-5}$ M NAA and $10^{-4}$ M BA, showing 3.08 g of callus fresh weight determined at 30 days after incubation. The highest chlorophyll content of callus was obtained in the LS medium supplemented with $10^{-5}$M NAA and $10^{-6}$ M BA, showing 28.42${\mu}g/g$. In both light and dark conditions, callus induction increased as the concentration of sucrose increased from 0.5%to 3.0%. In particular, in the light condition, the greatest callus induction was made in the LS medium supplemented with 2% sucrose. On the other hand, the heighest chlorophyll content was observed at 0.5% sucrose and chlorophyll wasn't induced in dark condition. The chlorophyll content of callus cultured in LS medium containing 0.75% sucrose was similar to 1% sucrose, as far as the chlorophyll content of callus was concerned.

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Screening of Anti-inflammatory Compound-producing Wild Yeasts and Their Microbiological Characteristics (항염증 물질 생산 능력이 우수한 야생효모의 선별 및 이들의 균학적 특성)

  • Bae, Sang-Min;Han, Sang-Min;Lee, Jong-Soo
    • The Korean Journal of Mycology
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    • v.45 no.3
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    • pp.212-223
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    • 2017
  • To screen for potent anti-inflammatory compound-producing yeasts, we evaluated nitric oxide production inhibitory activities in RAW 264.7 macrophage cells using cell-free extracts from 182 non-pathogenic yeasts. Rhodotorula graminis YJ36-1 and Meyerozyma guilliermondii YJ34-2 showed high inhibitory activities of 57.4% and 47.0%, respectively. The microbiological characteristics of these yeasts were investigated. Rhodotorula graminis YJ36-1 formed ascospores and pseudomycelium. This species grew well at $25^{\circ}C$ in yeast extract-peptone-dextrose (YPD) medium, vitamin-free medium, and 5% NaCl-containing YPD medium. Meyerozyma guilliermondii YJ34-2 was an asporogenous yeast and did not form pseudomycelium. This strain also grew well at $30^{\circ}C$ in YPD medium, vitamin-free medium, and 5% NaCl-containing YPD medium.

Construction of a Transgenic Plant to Develop a New Method for the Isolation of Calmodulin-Binding Proteins (새로운 방법을 이용한 칼모둘린 결합 단백질 분리를 위한 형질 전환 식물체의 구축)

  • Kim, Sun-Ho;Lee, Kyung-Hee;Kim, Kyung-Eun;Jung, Mi-Soon;Lim, Chae-Oh;Lee, Shin-Woo;Chung, Woo-Sik
    • Journal of Life Science
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    • v.17 no.9 s.89
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    • pp.1177-1181
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    • 2007
  • Calmodulin (CaM), a ubiquitous calcium-binding protein, regulates diverse cellular functions by modulating the activity of a variety CaM-binding proteins (CaMBPs). Because eukaryotes have multiple CaMBPs, it is important to isolate and characterize them in different tissues and conditions. So far a number of CaMBPs have been identified through classical screening methods. Many classes of proteins have been predicted to bind CaMs based on their structural homology with already known targets. In an effort to develop a method for large-scale analysis of CaMBPs in Arabidopsis, we have generated a transgenic plants overexpressing AtCaM2-GFP. We performed protein pull-down assay to test whether exogenously expressed AtCaM2-GFP proteins can interact with CaMBPs. The exogenously expressed AtCaM2-GFP could strongly interact with a CaMBP, AS1 protein. This result suggests that AtCaM2-GFP in transgenic plants may interact with many CaMBPs in plant cell. Therefore, we will be able to isolate kinds of CaMBPs by using these transgenic plants in many different tissue and environments.

Effects of 5-azacytidine, a DNA methylation inhibitor, on embryogenic callus formation and shoot regeneration from rice mature seeds (벼 성숙종자로부터 배상체 캘러스 형성 및 식물체 재분화에 DNA methylation 억제제인 5-azacytidine의 영향)

  • Lee, Yeon-Hee;Lee, Jung-Sook;Kim, Soo-Yun;Sohn, Seong-Han;Kim, Dool-Yi;Yoon, In-Sun;Kweon, Soon-Jong;Suh, Seok-Chul
    • Journal of Plant Biotechnology
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    • v.35 no.2
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    • pp.133-140
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    • 2008
  • The modification of DNA and histone plays an important role for gene expression in plant development. The objective of this research is to observe the effects of methylation on the gene expression during dedifferentiation from rice mature seeds to callus and differentiation from callus to shoots. The embryogenic callus with ability to shoot regeneration was not induced on the N6A medium supplemented with 5-azacytidine and abnormal callus with brown color was formed. When the normal rice callus was placed on the regeneration MSRA medium supplemented with 5-azacytidine, the shoot regeneration was inhibited. The results showed that 5-azacytidine, DNA demethylating agent, had negative effects on normal embryogenic callus formation and shoot regeneration. This suggested that DNA methylation of some genes was required for normal cell dedifferentiation and differentiation in tissue culture. The microarray and $GeneFishig^{TM}$ DEG screening were used to observe the gene transcript profile in callus induction and regeneration on N6A (N6 medium + 5-azaC) and MSRA (MS regeneration medium + 5-azaC). Subsets of genes were up-regulated or down-regulated in response to 5-azaC treatments. The genes related with epigenetic regulation, electron transport, nucleic acid metabolism and response to stress were up and down regulated. The different expression of some genes (germin like protein etc.) during callus induction and shoot regeneration was confirmed using RT-PCR and northern blot analysis.

Immune Function-enhancing Effects of Bojungikkitanggami-bang (보중익기탕가미방의 면역기능 증진 효과)

  • Lee Sang Hun;Lee Seung Eon;Lee Si Hyeong;Shin Jo Young
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.18 no.2
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    • pp.528-533
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    • 2004
  • The immune system acts to protect the host from infectious agents that exist in the environment and from other noxious insults. The immune system has two functional divisions: the innate and the acquired. Both components involve various factors such as cytokines. A number of methodologies exist to assess aspects of immune function. There are large inter-individual variations in many immune functions even among the healthy. Genetics, age, gender, smoking habits, habitual levels of exercise, alcohol consumption, diet, stage in the female menstrual cycle, stress, history of infections and vaccinations, and early life experiences are likely to be important contributors to the observed variation. While it is clear that individuals with immune responses significantly below 'normal' are more susceptible to infectious agents and exhibit increased infectious morbidity and mortality, it is not clear how the variation in immune function among healthy individuals relates to variation in susceptibility to infection. Oriental medicine is an important factor contributing to immune competence. The author investigated the immune enhancement effects of Bojungikkitanggami-bang (BITB). The forced swimming test (FST) has been used as a screening model for new immune enhancement agents. In the present study, the author investigated the effects of BITB on FST and blood biochemical parameters related to fatigue, glucose (Glc); blood urea nitrogen (BUN); lactate dehydrogenase (LDH); creatinine; and total protein (TP). The author found that BITB (1 g/kg) significantly reduced the immobility time in the FST compared to the control. In addition, the contents of Glc, LDH, BUN, TP in the blood serum were increased in BITB (1g/kg)-fed group. Also, the author investigated the effects of BITB on the production of cytokines in human T-cell line, MOLT-4 cells. BITB (1 mg/ml) significantly increased the interferon (IFN)-vproduction compared with media control (about 2.2-fold for IFN-γ) at 24 h. However, BITB has not affect the production of IL-2 and IL-4. In addition, BITB increased the protein expression level of IFN-γ in MOLT-4 cells. Thus, BITB may have therapeutic value in generating or enhancing immune function in a clinical setting.

Real-time Micro-algae Flocculation Analysis Method Based on Lens-free Shadow Imaging Technique (LSIT) (렌즈프리 그림자 이미징 기술을 이용한 실시간 미세조류 응집현상 분석법)

  • Seo, Dongmin;Oh, Sangwoo;Dong, Dandan;Lee, Jae Woo;Seo, Sungkyu
    • Journal of the Korean Society for Marine Environment & Energy
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    • v.19 no.4
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    • pp.341-348
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    • 2016
  • Micro-algae, one of the biological resources for alternative energy, has been heavily studied. Among various methods to analyze the status of the micro-algae including counting, screening, and flocculation, the flocculation approach has been widely accepted in many critical applications such as red tide removal study or microalgae resource study. To characterize the flocculation status of the micro-alga. A traditional optical modality, i.e., photospectrometry, measuring the optical density of the flocs has been frequently employed. While this traditional optical method needs shorter time than the counting method in flocculation status analysis, it has relatively lower detection accuracy. To address this issue, a novel real-time micro-algae flocculation analysis method based on the lens-free shadow imaging technique (LSIT) is introduced. Both single cell detection and floc detection are simultaneously available with a proposed lens-free shadow image, confirmed by comparing the results with optical microscope images. And three shadow parameters, e.g., number of flocs, effective area of flocs, and maximum size of floc, enabling quantification of the flocculation phenomenon of micro-alga, are firstly demonstrated in this article. The efficacy of each shadow parameter is verified with the real-time flocculation monitoring experiments using custom developed cohesive agents.

Fermentation Studies on Pseudomonas aeruginosa Producing Antifungal Secondary Metabolite, PAFS. (항진균물질을 생합성하는 Pseudomonas aeruginosa의 배양생리적 특성 연구)

  • 송성기;윤권상;정용섭;전계택
    • Microbiology and Biotechnology Letters
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    • v.32 no.1
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    • pp.52-59
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    • 2004
  • When both fructose and galactose were added to a production medium as carbon sources, the productivity of PAFS (Psedomonas Antifungal Substance) biosynthesized by Pseudomonas aeruginosa was observed to be reduced significantly due to the well-known phenomenon of catabolite repression. In order to overcome this phenomenon by use of fermentation bioprocess, fed-batch cultivation method was examined. In addition, a high producer mutant strain, AP-20 obtained by a rational screening method was tested for its productivity of PAFS in both batch and fed-batch fermentation processes. Notably fed-batch operation showed approximately 4 fold higher PAFS productivity than traditional batch operation process. It was appeared that galactose was utilized principally for the cell growth of Pseudomonas aeruginosa whereas large portion of fructose was used for the biosynthesis of PAFS. Furthermore it was observed that composition and feeding rate of production media should be optimized even in the fed-batch fermentation bioprocess. As an example, very slow feeding of carbon sources gave rather negative effect on the production of PAFS due to significant limitation of carbon and energy sources available for the producer microorganism.

In Vitro Screening for Compounds Derived from Traditional Chinese Medicines with Antiviral Activities Against Porcine Reproductive and Respiratory Syndrome Virus

  • Cheng, Jia;Sun, Na;Zhao, Xin;Niu, Li;Song, Meiqin;Sun, Yaogui;Jiang, Junbing;Guo, Jianhua;Bai, Yuansheng;He, Junping;Li, Hongquan
    • Journal of Microbiology and Biotechnology
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    • v.23 no.8
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    • pp.1076-1083
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    • 2013
  • Seventeen compounds derived from traditional Chinese medicines (TCMs) were tested for their antiviral activity against porcine reproductive and respiratory syndrome virus (PRRSV) in vitro. Visualization with the cytopathologic effect (CPE) assay and the 3-(4, 5-dimethyithiazol-2-yl)-2,5-diphenyltetrazolium bromide test were used to determine the 50% cytotoxic concentration ($CC_{50}$) and 50% effective concentration ($EC_{50}$) in cultured Marc-145 cells. Among the tested compounds, chlorogenic acid and scutellarin showed potential anti-PRRSV activity. The $EC_{50}$ values were $270.8{\pm}14.6{\mu}g/ml$ and $28.21{\pm}26.0{\mu}g/ml$ and the selectivity indexes were >5.54 and 35.5, respectively. The time-of-addition and virucidal assay indicated that the anti-PRRSV activity of the two compounds could be due to their inhibiting the early stage of virus replication and/or inactivating the virus directly. The inhibition of the virus attachment was not observed in the adsorption inhibition assay. The inhibition ratios of chlorogenic acid and scutellarin were, respectively, 90.8% and 61.1% at the maximum non-cytotoxic concentrations. The results have provided a basis for further exploration of their antiviral properties and mechanisms in vivo. We believe that the chlorogenic acid and scutellarin have a great potential to be developed as new anti-PRRSV drugs for clinical application.

Gene Transfer into Pig and Goat Fetal Fibroblasts by Co-transfection of tPA Transgene and $Neo^r$ Gene

  • Kim, Bae-Chul;Han, Rong-Xun;Kim, Myung-Yoon;Shin, Young-Min;Park, Chang-Sik;Jin, Dong-Il
    • Reproductive and Developmental Biology
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    • v.33 no.2
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    • pp.107-111
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    • 2009
  • The transfection efficiency of a transgene into pig and goat fetal fibroblast cells (PFF and GFF, respectively) was tested using co-transfection of a human tissue-type plasminogen activator (tPA) transgene and neomycin-resistant ($Neo^r$) gene, followed by G418 selection. To initially test G418 resistance, GFF and PFF were incubated in culture medium containing different concentration of G418 for 2 weeks, and cell survival was monitored over time. Based on the obtained results, the concentrations chosen for G418 selection were 800 ug/ml and 200 ug/ml for GFF and PFF, respectively. For co-transfection experiments, the pBC1/tPA and $Neo^r$ vectors were co-transfected into GFF and PFF ($1{\times}10^6$ cells in each case) using the FuGENE6 transfection reagent, and resistant colonies were obtained following 14 days of G418 selection. We obtained 96 and 93 drug-resistant colonies of GFF and PFF, respectively, only 54 and 39 of which, respectively, continued proliferating after drug selection. PCR-based screening revealed that 23 out of 54 analyzed GFF colonies and 5 out of 39 analyzed PFF colonies contained insertion of the tPA gene. Thus, the experimentally determined transfection efficiencies for tPA gene co-transfection with the $Neo^r$ gene were 42.6% for GFF and 12.8% for PFF. These findings suggest that co-transfection of a transgene with the $Neo^r$ gene can aid in the successful integration of the transgene into fetal fibroblast cells.

Anti-inflammatory and Anti-oxidative Activities of Flavonoids Extracted from Dendranthema indicum Flowers in Jeju Island (제주 자생 감국 꽃 추출물 유래 flavonoid 화합물의 항산화 및 항염 활성)

  • Hyun, Ju Mi;Jo, Yeon Jeong;Kim, Yun Beom;Park, Sung-Min;Yoon, Kyung-Sup;Lee, Nam Ho
    • Journal of the Korean Applied Science and Technology
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    • v.36 no.4
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    • pp.1259-1267
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    • 2019
  • Anti-inflammatory and anti-oxidative activities were examined on the extract of Dendranthema indicum (D. indicum) flowers. The flowers were extracted two times for 24 h each with 70% ethanol. Upon the biological activities screening, the ethanol extract exhibited potent free radical scavenging activities and inhibited the production of nitric oxide on LPS-induced RAW264.7 macrophages effectively without causing cell toxicity. Further purification by medium pressure liquid hromatography (MPLC) and identification of the isolates led to identification of cynaroside (1) and apigetrin (2). The chemical structures of the isolated compounds were elucidated based on spectroscopic data including nuclear magnetic resonance (NMR) spectra, as well as comparison of the data to the literature values. Also, the quantitative analysis of the compounds was perfromed by high-performance liquid chromatography (HPLC). The isolates 1 and 2 were determined to inhibite the nitric oxide (NO) production dose-dependently. Based on these results, it was suggested that D. indicum extract could be useful as anti-inflammatory agents in cosmetics applications.