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植物染色质免疫共沉淀方法 被引量:4
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作者 李东明 宋渊 安黎哲 《草业科学》 CAS CSCD 北大核心 2014年第4期659-667,共9页
染色质结构的动态变化和基因的适时适量表达,在植物生长发育过程中起着非常重要的作用。研究染色质结合蛋白和DNA的时空结合关系,对认识这些复杂的生命过程有重大意义。染色质免疫共沉淀(Chromatin Immunoprecipitation)就是在体内染色... 染色质结构的动态变化和基因的适时适量表达,在植物生长发育过程中起着非常重要的作用。研究染色质结合蛋白和DNA的时空结合关系,对认识这些复杂的生命过程有重大意义。染色质免疫共沉淀(Chromatin Immunoprecipitation)就是在体内染色质环境下研究蛋白质和DNA结合关系的方法,本研究利用先提核再甲醛交联的方法,详细叙述了该方法的6个主要步骤。通过对拟南芥(Arabidopsis thaliana)野生型Col-0和突变体met1中组蛋白H3K9甲基化在At4g03870转座子上差异的研究,认为染色质免疫共沉淀方法可行、稳定,此方法不仅适用于单个基因与蛋白结合关系的研究,还适用于特异蛋白结合位点的全基因组学研究。 展开更多
关键词 染色质结构 提细胞核 超声波破碎 染色体免疫共沉淀
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Prunella vulgaris L. extract improves cellular immunity in MDR-TB challenged rats 被引量:3
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作者 Lu Jun Qin Rui +1 位作者 Ye Song Yang Mei 《Journal of Medical Colleges of PLA(China)》 CAS 2011年第4期230-237,共8页
Objective: To study the effect of the extract of Prunella vulgaris L. on multiple drugs resistant bacillus tuberculosis (MDR-TB). Methods: Experimental animal model in rats was induced by MDR-TB. Normal group mode... Objective: To study the effect of the extract of Prunella vulgaris L. on multiple drugs resistant bacillus tuberculosis (MDR-TB). Methods: Experimental animal model in rats was induced by MDR-TB. Normal group model group and Prunella vulgaris L. group were set up. The contents of IFN-7, IL-4, IL-10 and IL-12 were examined by ELISA. Their genome mRNAs were extracted, the target genes were amplified by PCR. RT-PCR was used to detect the mRNA levels of them. Results: The content of IFN-q, of the extract of Prunella vulgaris L. group was 1.98±0.67 pg/ml, IL-4 was 6.47±1.46 pg/ml, IL-10 was 12.13±3.43 pg/ml and IL-12 was 3.02±0.86 pg/ml. Compared with the model group, Prunella vulgaris L. group was notable difference in serum IFN-γ, IL-12 and IL-10 (P〈0.05). The mRNA levels of IFN-γ, IL-12 increased and IL-10 decreased obviously, the differences were quite significant (P〈0.05), but IL-4 had no obvious change. Conclusion: The extract of Prunella vulgaris L. can enhance the cellar immunological function in rats from up-regulation of the level of genetic transcription, accordingly provide the theory basis of healing of tuberculosis with it. 展开更多
关键词 Prunella vulgaris L. EXTRACT Multiple drugs resistant bacillus tuberculosi CYTOKINE Immunological function RATS
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Nuclear assembly of purified Crythecodinium cohnii chromosomes in cell-free extracts of Xenopus laevis eggs
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作者 LIU XIAO LING YAN SHEN +1 位作者 ER JIE CHEN ZHONG HEZHAI(E-mail: zhaizh@plum.lsc.pku.edu.cn Tel: 86-10-62754293 Fax: 86-10-62751526)(College of Life Sciences, Peking University, Beijing 100871,China) 《Cell Research》 SCIE CAS CSCD 2000年第2期127-137,共11页
Incubation of dinoflagellate Crythecodinium cohnii chromosomes in cytoplasmic extracts of unfertilized Xenopus laevis eggs resulted in chromosomes decondensation and recondensation, nuclear envelope assembly, and nucl... Incubation of dinoflagellate Crythecodinium cohnii chromosomes in cytoplasmic extracts of unfertilized Xenopus laevis eggs resulted in chromosomes decondensation and recondensation, nuclear envelope assembly, and nuclear reconstitution.Dinoflagellate Crythecodinium cohnii is a kind of primitive eukaryote which possesses numerous permanently condensed chromosomes and discontinuous double-layered nuclear membrane throughout the cell cycle. The assembled nuclei, being surrounded by a continuous double membrane containing nuclear pores and the uniformly dispersed chromatin fibers are morphologically distinguishable from that of Dinoflagellate Crythecodinium cohnii. However, incubation of dinoflagellate Crythecodinium cohnii chromosomes in the extracts from dinoflagellate Crythecodinillm cohnii cells does not induce nuclear reconstitution. 展开更多
关键词 Dinoflagellate Crythecodinium cohnii CHROMOSOME nuclear reconstitution cell-free system Xenopus laevis egg extracts
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Association of DNA with nuclear matrix in in vitro assembled nuclei induced by rDNA from Tetrahymena shanghaiensis in Xenopus egg extracts
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作者 CHEN YING BO ZHANG +1 位作者 XIU FEN LI ZHONG HE ZHAI(Department of Cell Biology and Genetics, College of Life Sciences, Peking University, Beijing 100871) 《Cell Research》 SCIE CAS CSCD 1997年第1期107-117,共11页
The nuclei assembled from exogenous DNA or chromatin in egg extracts resemble their in vivo counterparts in many aspects. However, the distribution pattern of DNA in these nuclei remains unknown. We introduced rDNA fr... The nuclei assembled from exogenous DNA or chromatin in egg extracts resemble their in vivo counterparts in many aspects. However, the distribution pattern of DNA in these nuclei remains unknown. We introduced rDNA from the macronuclei of Tetrabymena into Xenopus cellfree extracts to examine the association of specific DNA sequences with nuclear matrix (NM) in the nuclei assembled in vitro. Our previous works showed the 5’NTS (nontranscription sequences) of the rDNA specifically bind to the NM system in the macronuclei. We show now the rDNA could induce chromatin assembly and nuclear formation in Xenopus cell-free system. When we extracted the NM system and compared the binding affinity of different regions of rDNA with the NM system, we found that the 5’NTS still hold their binding affinity with insoluble structure of the assembled nuclei in the extracts of Xenopus eggs. 展开更多
关键词 Nuclear assembly nuclear matrix Xenopus egg extracts Tetrahymena rDNA
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Bone Marrow Mononuclear Cell Extraction. Does the Operator Performance Affect Processing Efficacy?
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作者 Jakobsons Eriks Cakstina Inese +9 位作者 Erglis Kristaps Erglis Martins Pirtniece Liene Elina Krumina NarbuteInga Briede Ieva Jegere Sanda Smolova Rita Muiznieks Indrikis Erglis Andrejs 《Journal of Pharmacy and Pharmacology》 2016年第5期192-198,共7页
Different cell populations from bone marrow are used in various clinical trials for cardiac diseases during last decade. Four clinical studies are on going in our institution and enrol patients with cardiac diseases, ... Different cell populations from bone marrow are used in various clinical trials for cardiac diseases during last decade. Four clinical studies are on going in our institution and enrol patients with cardiac diseases, coronary disease and type 2 diabetes, patients with osteoarthritis. Density gradient is used to separate bone marrow mononuclear cells. Cell processing looses are significant. To find out critical control points we analysed processing process and differences in cell yields between operators performing cell extraction. Bone marrow mononuclear cells were isolated using Ficoll density gradient centrifugation. Cells were counted using flow cytometry for mononuclear cell total counts, CD34+ population count and viability analysis. The patients who underwent bone marrow aspiration followed by cell isolation received cell suspension for transplantation. Two cells processing for separate patients were performed at once. Same standard operation procedures were applied. Processing looses between operators performing cell extraction were analysed. Bone marrow samples from eight patients were processed. Mononuclear cells were extracted. Operator performances were compared. Operator A average bone marrow mononuclear cell yield in starting material was 9,97 ± 9,98 %, CD34+ population yield 75,46±79,67%. Operator B average bone marrow mononuclear cell yield in starting material was 24,68 ± 14,8 %, CD34+ population yield 70,42 ±44.84%. Operator A average cell viability in starting material was 45,24 ± 9,55%, after cell processing 42,96 ± 23,66 %. Operator B average cell viability in starting material was 49,85 ± 5,48%, after cell processing 69,52 ± 6,65 %. 展开更多
关键词 Bone marrow mononuclear cells cell yield cell extraction cell count
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