The Construction of a Vector for Endosperm-specific Expression of IPT Gene and It's Transformation in Rice

为研究细胞分裂素在水稻籽粒发育中的作用,构建了P1300-P Pg-5α-IPT-nOS植物表达载体,并对水稻进行遗传转化。以水稻品种9311为材料,采用PCr法获得种子中特异表达的Pg-5α基因启动子,并将此启动子与P1300相连接,构建P1300-P Pg-5α载体,用nCOⅠ和SPEⅠ双酶切P Sg516,获得IPT-nOS核酸片段,然后将此核酸片段插入到P1300-P Pg-5α中,构建P1300-P Pg-5α-IPT-nOS植物表达载体。以水稻品种日本晴愈伤组织为材料,采用农杆菌介导的方法进行遗传转化。成功构建了植物表达载体P1300-P Pg-5α-IPT-nOS,获得了阳性率为81.3%转IPT基因群体。获得转基因株系后,将为进一步筛选高效表达株系及观察其籽粒生长发育过程提供基础。In order to specifically express IPT gene in the endosperm of developing rice seed,the binary vector p1300-p PG-5α-IPT-Nos was constructed,firstly,the promoter of PG-5α was obtained by PCR amplification from rice genomic DNA( variety: 9311) and p1300-p PG-5α was constructed by inserting p PG-5α into p1300,then the IPT-Nos fragment was obtained by cutting p SG516 with Nco Ⅰ and Spe Ⅰ.After IPT-Nos inserted into p1300-p PG-5α,p1300-p PG-5α-IPT-Nos was constructed.p PG-5α-IPT-Nos was transformated into Oryza sativa( subsp.japonicacv.Nipponbare) cells by agrobacterium mediated method and regenerative seedlings were obtained.The results of PCR showed that about 81.3% seedlings were positive.唐山师范学院博士基金项目(09A01); 农业部“948”项目(2011-G1(3)-07); 河北省科技计划项目(13396401D

Expression and Localization of hnRNP A2/B1 during Differentiation of Human Osteosarcoma MG-63 Cells Induced by HMBA

背景与目的:核基质蛋白的差异表达与细胞癌变和增殖分化调控关系密切。本研究观察了hnRNP A2/B1在诱导分化处理前后人成骨肉瘤MG-63细胞核基质中的存在和分布,及其与Actin、Prohibitin的共定位关系。方法:选择性抽提经环六亚甲基双乙酰胺(hexamethylene bisacetamide,HMBA)诱导处理前后的MG-63细胞核基质,并运用双向电泳、质谱分析、蛋白质印记杂交、免疫荧光、激光共聚焦等技术检测hnRNP A2/B1在核基质中的表达与定位变化,及其与相关蛋白的共定位关系。结果:双向电泳及蛋白质印迹杂交结果证实了hnRNP A2/B1存在于MG-63细胞核基质蛋白组分中,并在HMBA处理后细胞核基质中表达下调;免疫荧光显微镜观察显示hnRNP A2/B1定位在核基质上,经HMBA处理后hnRNP A2/B1表达减弱。激光共聚焦显微镜观察结果显示,hnRNP A2/B1与细胞核基质蛋白组分Actin、细胞增殖相关调控因子Prohibitin具有共定位关系,但在诱导处理后细胞内的共定位关系减弱。结论:hnRNP A2/B1在MG-63细胞诱导分化过程中的表达分布,及其与Actin、Prohibitin的共定位关系的改变对MG-63细胞分化具有重要影响,值得进一步探索和研究。BACKGROUND & OBJECTIVE: The differentially expressed nuclear matrix proteins have great effects on canceration and regulation of cell differentiation. This study was to explore the existence and distribution of ribonucleoprotein hnRNP A2/B1 in nuclear matrix and its co-localization with Actin and Prohibitin in human osteosarcoma MG-63 cells before and after hexamethylene bisacetamide (HMBA) treatment. METHODS: The nuclear matrix of MG-63 cells before and after treatment of HMBA were selectively extracted. The expression and localization of hnRNP A2/B1 in nuclear matrix were detected by 2-D PAGE, MALDI-TOF-MS, Western blot, and immunofluorescent staining. The co-localization of hnRNP A2/B1 with Actin and Prohibitin was observed under laser scanning confocal microscope (LSCM). RESULTS: hnRNP A2/B1 was detected in the component of nuclear matrix proteins of MG-63 cells by Western blot and immunogold staining and its expression was decreased after treatment of HMBA. hnRNP A2/B1 was located in the nuclear matrix, and its expression was weakened after HMBA treatment. hnRNP A2/B1 was co-localized with Actin or Prohibitin in MG-63 cells, while the co-localization relationship was weakened during differentiation of MG-63 cells. CONCLUSIONS: hnRNP A2/B1 is a kind of nuclear matrix protein, and localizes in the nuclear matrix. The distribution and expression of hnRNP A2/B1 and its co-localization with Actin and Prohibitin play important roles during the differentiation of MG-63 cells.国家自然科学基金项目(No.30470877)~

圣·约翰草提取物片治疗抑郁障碍专家共识

结合循证医学证据,本文归纳圣·约翰草提取物片治疗抑郁障碍的专家共识

The contents and distribution of phosphorus in natural colloids

国家教育部高等学校骨干教师计划基金资助项目;; 国家自然科学基金重大研究计划重点项目资助(90411016

ABA促进针叶树体细胞胚胎分化

文章介绍ABA促进针叶树体细胞胚胎分化调控机制和应用的研究进展

Determination of hydrogen peroxide in food

利用猪血红蛋白(PHb)的催化特性,以3,3',5,5'-四甲基联苯胺(TMb)为底物,建立了一种催化光度终止法测定食品中残留过氧化氢(H2O2)的方法。实验考察了终止剂的类型和浓度、反应时间、PH、温度、TMb浓度对H2O2检测的影响。在选定的反应条件下,体系吸光值与H2O2浓度在1.46x10-6~2.91x10-5MOl/l范围内呈良好的线性关系,线性相关系数为0.9958,方法对H2O2的检出限为5x10-7MOl/l,测定标准偏差为4.9%,样品的加标回收率为92.9%~103%,利用该方法进行了实际样品中的H2O2分析,结果与国标法无显著性差异。A new spectrophotometric method was developed for the determination of hydrogen peroxide(H2O2) in foods using 3,3′,5,5′-tetramethylbenzidine(TMB) as a terminating agent in the catalytic reaction between hemoglobin porcine(pHb) and H2O2.The influence of terminating agents,reaction time,pH,temperature and the concentration of TMB was discussed.Under the optimal experimental conditions,the absorbance value was linearly correlated to H2O2 concentration in the range of 1.46×10-6～2.91×10-5 mol/L with the correlation coefficient of 0.9958.The detection limit was found to be 5×10-7 mol/L and RSD was 4.9 %.The recoveries in the range of 92.9 %～103 % were obtained.The determination results corresponded well with those by the national standard method

浅海井口抗冰装置结构计算的有限单元法

对辽东湾浅海单井井口保护装置采用隔水套管直接抗冰的可行性进行了研究。采用多种三维混合单元建立了一典型井口装置的有限元计算模型，该模型利用有限差分的原理真实地反映了地层对结构的约束。计算结果表明，在辽东湾浅海的60cm单层平整冰作用下，结构的位移和应力都在许可范围之内，最大位移为结构的顶部，最大应力发生在隔水套管与表层套管的上部连接环板上，而隔水套管内加固的水泥层对增加结构的强度、抵抗外载荷的作用起了不可忽视的作用。计算结果和现场两年多的应用表明，用增厚的隔水套管直接抗冰是可行的

新疆伊犁河流域生态服务价值对土地利用变化的响应

以伊犁河流域为研究对象,运用GIS手段和生态经济学方法,采用Costanza生态系统服务价值计算公式,参照谢高地等人的中国陆地生态系统服务单位面积价值,结合敏感度分析,探讨了伊犁河流域1985-2005年土地利用和生态系统服务价值的变化特征。研究结果表明:1985-2005年间伊犁河流域土地利用处于发展阶段,土地利用程度呈增大趋势。研究期内生态系统服务价值总体有所上升,从482.83×108元上升到506.44×108元,增加了23.61×108元:水域和耕地对研究区总体服务功能变化的贡献率最大,而牧草地生态服务功能的下降对研究区总体生态服务功能的增加具有较大负贡献。研究区生态经济的发展已处于协调水平的边缘,需要加强对生态环境的保护力度