본 연구에서는 FT-IR 스펙트럼데이터의 다변량 통계분석 기법을 활용하여 인도수집 옥수수 계통 및 품종으로부터 단백질 함량이 높은 옥수수를 신속하게 선발할 수 있는 선발체계를 확립함과 동시에 lysine과 tryptophan의 함량분석을 목적으로 연구를 수행하였다. 총 48시료의 인도수집 옥수수 계통 및 품종과 국내산 품종을 이용하여 종자로부터 FT-IR 스펙트럼을 조사하였으며, 무작위로 선발된 24시 료를 대상으로 총 단백질 함량을 조사하였다. 대조구로 사용한 광평옥 모계(GPO1)의 경우 단백질 함량 이 9.34 ± 0.3mg/g dw인데 비하여 H4 계통의 경우 단백질 함량이 10.26 ± 0.5mg/g dw로 48개 옥 수수 시료 중에서 가장 높게 나타났다. 특히 옥수수 H4, H6, H11, 그리고 H12 계통의 경우 총 단백질 함량이 각각 10mg/g dw 이상으로 측정되어 광평옥 모계(9.34mg/g dw)와 부계(9.36mg/g dw) 및 이 들의 F1(9.14mg/g dw)보다 총 단백질 함량이 높은 계통으로 판명되었다. Cross-validation test에서 옥수수 종자 내 총 단백질 함량예측 PLS regression model의 regression coefficient(R2) 는 0.77로 비교적 정확하게 총 단백질 함량예측이 가능한 것으로 나타났다. 따라서 본 PLS regression model을 이용하여 단백질 함량이 높은 사일리지 옥수수 계통의 선발이 가능할 것으로 기대되며, 더 나아가 다양 한 옥수수 계통의 신속한 대사체 수준 평가가 가능할 것으로 예상된다.
The use of high throughput screening (HTS) in drug development is principally for the selection new drug candidates or screening of chemical toxicants. This system minimizes the experimental environment and allows for the screening of candidates at the same time. Umbilical cord-derived stem cells have some of the characteristics of fetal stem cell and have several advantages such as the ease with which they can be obtained and lack of ethical issues. To establish a HTS system, optimized conditions that mimic typical cell culture conditions in a minimal space such as 96 well plates are needed for stem cell growth. We have thus established a novel HTS system using human umbilical cord derived-mesenchymal stem cells (hUC-MSCs). To determine the optimal cell number, hUC-MSCs were serially diluted and seeded at 750, 500, 200 and 100 cells per well on 96 well plates. The maintenance efficiencies of these dilutions were compared for 3, 7, 9, and 14 days. The fetal bovine serum (FBS) concentration (20, 10, 5 and 1%) and the cell numbers (750, 500 and 200 cells/well) were compared for 3, 5 and 7 days. In addition, we evaluated the optimal conditions for cell cycle block. These four independent optimization experiments were conducted using an MTT assay. In the results, the optimal conditions for a HTS system using hUC-MSCs were determined to be 300 cell/well cultured for 8 days with 1 or 5% FBS. In addition, we demonstrated that the optimal conditions for a cell cycle block in this culture system are 48 hours in the absence of FBS. In addition, we candidates using our HTS system which demonstrates the feasibility if using hUC-MSCs for this type of screen. Moreover, the four candidate compounds can be tested for stem cell research application.
The purpose of this study was to establish a system for plant fluorescence image acquisition and to verify the possibility of plant fluorescence image analysis as a non-destructive method to screen the salt tolerance of soybean (Glycine max). Two-weeks-old seedlings of soybean at the V1 growth stage were treated with 0, 50, and 100 mM of NaCl for salt stress and plant fluorescence images were taken by CCD camera (EOS-600D, Canon, Japan) equipped with band pass filter (XNiteBPB, LPD LLC, USA) at 0, 15, 30, 60, 120 and 240 second after blue light exposure at 1 day after treatment. Red color intensity was extracted using MatLab 8.1 (The MathWorks Inc., USA) for estimation of plant fluorescence intensity. Red color intensity of soybean image decreased 0 (F0-10) to 240 (F240-250) second after blue light exposure irrespective of NaCl concentration, while F0-10/F240-250 decreased with NaCl concentration, resulting in significant relationship with plant fluorescence (Fv/Fm) and salt stress intensity. Therefore, our results suggest that our plant fluorescence image acquisition and analysis methods can be a part of high-throughput screening system for salt tolerance of soybean varieties
The protein in soybean seeds accounts for approximately 40% of the dry seed weight. Two major storage proteins, 7S and 11S, constitute 70-80% of the total storage proteins in the seeds. In this study, the variation of total soluble protein extracts from 1152 soybean landraces that have been collected from South Korea were studied using high-throughput screening method with HT Protein Express Labchip (Caliper Life Sciences, Inc.). Seven distinct protein band patterns - four protein sub-units of 11S and three sub-units of 7S, were taken into account and their presence or absence were analyzed. Among the 1152 landraces, 525 genotypes were identified as lacking lipoxygenase, 255 lacking α1 subunit, 680 lacking α subunit, 169 lacking β subunit, 140 lacking acidic, 114 lacking Kunitz Trypsin Inhibitor (KTi) and 199 lacking basic protein patterns. The high-throughput protein analysis is helpful in screening a large number of populations with less time and minimum labor. The selected genotypes with low amounts or lacking of anti-nutritional factors such as trypsin inhibitor, lipoxygenase and α subunit would be used for future breeding purpose of quality improvement in soybean protein.
식물 피노믹스 분야에서 초고속 대량선발이 가능하도록 만든 화상기술(imaging technology)을 온실자동화 기술, 이미지 촬영 및 분석기술 등으로 분류하여 개념을 정리하고, 화상기술을 개발 및 응용하고 있는 주요 연구기관의 현황을 소개하였다. 연구동향 파악을 위해 작물의 내재해성 검정, 병해충진단, 종자활력 검정, 수확후 관리, 생체리듬 연구 등 다양한 분야에서 응용되고 있는 사례들을 살펴보았다. 향후 열 화상, 형광 화상 기술을 UV-induc
The effects of angiogenesis inhibitor from the extract libraries of Korean and Chinese medicinal plants were investigated using a protein microarray chip. Protein chip was constructed by immobilization of integrin α5β1 on protein chip base plates and employed far screening active extracts that inhibit the integrin-fibronectin interaction from the extract libraries. The 100 extracts of medicinal plants were obtained from extract bank of National Institute of Crop Science, RDA. The 14 extracts among 100 extract libraries were shown efficient inhibition activity for the interaction between integrin-fibronectin. The medicinal plants of 14 extracts were Vitex negundo var. incisa (Lam.) C.B. Clarke, Epimedium koreanum Nakai, Cedrela sinensis A. Juss, Ipomea aquatica Forsk, Schisandra chinensis Baill, Pulsatilla koreana Nakai, Paeonia lactiflora Pall. var.hortensis Makino, Oenothera odorata, Allium chinense, Allium victorialis var. platyphyllum MAKINO, Polygonatum odoratum Druce var. pluriflorum Ohwi, Hosta lancifolia, Agrimonia pilosa L. var. japonica Nakai and Potentilla chinensis SER. The Paeonia lactiflora, Oenothera, and Agrimonia pilosa from these 14 extracts libraries were shown strong inhibition activity of integrin α5β1.
산화적인 스트레스(Oxidative stress)는 신경염증의 발병요인 중의 하나로 알려져 있다. 이에 본 연구는 약용곤충추출물을 이용하여 항산화 물질을 찾고자 초고속 적용가능한 스크리닝 방법을 적용하였다. 우선, 분자염증은 활성산소 관련의 물질과 밀접한 관련이 있으므로 이들의 억제를 동반하는 추출물을 먼저 선별하였다. 항산화 실험과 관련하여 DPPH (1,1-Diphenyl-2-picrylhydrazyl), FRAP(Ferric ion redu
천식과 관련하여 산화스트레스 (Oxidant stress)는 그 발병요인 중의 하나로 알려져 있다. 이에 본 연구는 농산물과 한약재를 이용하여 항산화 물질을 찾고자 하였으며, 시간과 비용의 단축을 위하여 HTS인 throughput screening)을 이용을 하였다. 항산화 실험과 관련하여서 DPPH(1,1-diphenyl-2-picrylhydrazyl), FRAP(ferric ion reducing antioxidant power), HO(h