人肿瘤坏死因子相关凋亡诱导配体基因植物表达载体的构建及瞬时表达

人肿瘤坏死因子相关凋亡诱导配体具有特异性诱导多种肿瘤细胞凋亡而对正常细胞无毒性的特点,被认为是肿瘤凋亡疗法较有希望的候选药物。植物生物反应器在药用蛋白质生产方面具备较大优势。构建了可溶性人肿瘤坏死因子相关凋亡诱导配体基因的植物表达载体,进行本氏烟(Nicotiana benthamiana)瞬时表达,并对目的蛋白的体外活性进行检测。结果显示,本氏烟叶片中目的蛋白平均表达量为68.60 pg/mg TSP;在浓度为200 pg/m L时,对NCI-H460细胞株的抑制率为28.75%。研究表明,经密码子优化的编码基因能够在受体植物中表达出有活性的目的蛋白,为开展目的基因的稳定转化工作奠定了基础。     

根癌农杆菌介导转化籼稻影响因素的研究

选用我国籼型杂交稻中的几个重要亲本 ,以携带有双元载体 p CAMBIA1 3 0 1的根癌农杆菌 EHA1 0 5为载体 ,以GUS基因的瞬间表达为依据 ,研究根癌农杆菌介导转化籼稻的影响因素 ,确立对转化频率有重要影响的几个条件。结果表明 :经短期预培养 (4～ 5d)的幼胚是较为理想的转化受体材料 ;在共培养基中添加较高浓度的乙酰丁香酮 (3 0 0～ 4 0 0μmol/ L)可提高 GUS基因的瞬间表达频率 ;以 CC培养基作为共培养后抗性愈伤筛选的基本培养基有利于抗性愈伤组织的筛选培养。按此确立的条件 ,在特青、龙特甫 (B)、珍汕 97(B)、盐恢 559等籼稻品种或亲本中的 GUS瞬间表达率高达 80 % ,抗性愈伤组织转化率高达 70 %     

减阻剂室外埋地评价环道及瞬态模型

针对减阻剂室内评价环道存在的问题,设计安装了管径159mm、长300m的室外埋地评价环道,其管径、长度、油品流速更接近于实际管道.该装置采用高压氮气作为动力源,测试段沿线布设高精度压力传感器和高频压力传感器,测试段出口设有超声波流量计,同时安装有耐压罐和常压罐.由于在环道评价试验中装置的压力不断变化,因此建立了试验过程氮气管路、油品管路、氮气罐、耐压罐的瞬态模型,使用VB语言编制了计算程序,用以计算不同粘度和初始压力下的油品流速和压力变化,雷诺数范围和试验时间,判断试验条件的可行性.     

农杆菌浸润瞬时表达分析番木瓜miR162a的研究

农杆菌浸润瞬时表达方法不仅在蛋白互作、鉴定基因沉默抑制子及基因功能研究方面得到广泛应用,目前还应用在小RNA的研究中.在对植物microRNA的研究中,农杆菌浸润方法通过将将转基因导入农杆菌浸润本生烟瞬时表达,可以快速、稳定地对植物microRNA进行检测和鉴定,样品在几天内就可分析完成,比获得稳定遗传的转基因有优势.笔者借鉴国外拟南芥microRNA瞬时表达分析的方法,克隆了番木瓜的miR62a前体序列连接到双元表达载体pSuper1300上构建成35S::cpa-miR162a质粒,导人农杆菌GV3101后浸润本生烟,浸润后48 h提取本生烟叶片总RNA后通过miRNA northern blot检测到miR162a,而本生烟和番木瓜自身则无法检测到低丰度的miR162a.本文介绍的农杆菌浸润瞬时表达方法能够在体外加工产生成熟的miRNA,该技术有助于对microRNA的结构-功能的相关性及靶标验证进一步研究.     

长输管道水力瞬变仿真

根据各种管道边界的瞬变方程和管道水击的连锁Ａｌｌｉｅｖｉ方法，针对具有实际元器件和边界的长输管道的不稳定流动，设计了基于特征线差分解法的分析流程和仿真计算程序。利用长输管道水击数值计算所得线路上各节点的瞬态值进行Ｂ样条曲线拟合，实现了水力瞬变过程中线路水力坡降线及某些水力参数变化情况的图形输出 。     

梅PmARF17克隆及其在花发育中与内源激素的调控模式

【目的】分析梅PmARF17的生物学功能,探究梅花发育进程中其表达丰度与内源激素动态变化的关系,为梅花发育的调控研究提供依据。【方法】以梅品种‘大嵌蒂’为试材,克隆PmARF17,利用生物信息学软件分析基因结构、系统进化及其与其他物种同源蛋白的差异;亚细胞定位确定PmARF17蛋白在细胞中作用的部位;以梅品种‘大嵌蒂’和‘龙眼’不同发育阶段的花芽、叶芽、花器官为试材,利用qRT-PCR检测PmARF17时空表达模式,通过UPLC法测定IAA、GA₃、ABA、ZT含量的动态变化,并与PmARF17的表达进行相关性分析;克隆PmARF17启动子,分析启动子的顺式作用元件,利用瞬时表达解析PmARF17与GA₃的调控模式。【结果】从梅品种‘大嵌蒂’中克隆得到PmARF17,系统进化树分析表明PmARF17蛋白与其他植物的ARF蛋白序列高度同源;亚细胞定位表明其作用于细胞核和细胞膜上;qRT-PCR表达和内源激素含量的相关性分析表明,PmARF17的表达与IAA含量的变化趋势没有明显的相关性。PmARF17在雌蕊完好花芽中的表达水平相对不完全花芽显著上调,而GA₃含量与PmARF17的表达趋势一致。ABA和ZT含量总体上与PmARF17的表达呈相反的趋势,表明两者可能抑制PmARF17的表达。PmARF17启动子含有GA顺式元件,且具有启动活性和组织表达特异性,在花瓣、雄蕊及根部特异表达。【结论】 PmARF17可能是梅花发育的正调控基因,促进梅雌蕊的正常发育。PmARF17的表达可能受到GA₃的正调控,其可能通过作用于雄蕊和花瓣,进而影响梅的雌蕊发育进程。     

Time dependence of hydraulic parameters estimation from transient analysis of mini disc infiltrometer measurements

The transient analysis of mini disc infiltrometer (MDI) measurements is an established method for characterising near-surface hydraulic characteristics of soils. The reliability of hydraulic characteristics obtained from transient analysis depends on the (1) adequacy of model, (2) adequacy of data, (3) measurement time and (4) measurement footprint. The measurement time dependence recommendations are reported only for a few soil textures, initially wet samples and tension infiltrometer (TI) with a higher measurement footprint than the MDI. This study investigated the adequacy of infiltration data (using cumulative linearization [CL] and differentiated linearization [DL]) and measurement time influence on the hydraulic parameters determined from the transient analysis of MDI measurements for six soil textures. The objective of the study is to identify suitable MDI measurement durations for different soil textures for the initially dry state, considering both adequacy of data and time fractionation (measurement time influence). The data adequacy time obtained from the DL (T_DL) was found to be 0.8 times less than the value obtained from CL (T_CL). The marginal difference in T_DL and T_CL had a significant influence on the determination of infiltration equation coefficient C₁ and negligible influence on coefficient C₂. The time fractionation procedure adopted for identifying adequate MDI measurement time (T_m) was found to be comparable based on sorptivity (S₀) and hydraulic conductivity (K₀). The average T_m was also comparable with T_DL and T_CL with a strong positive correlation. The C₂ values obtained based on T_m, T_DL, and T_CL were in better agreement than the corresponding C₁ values. The adequate MDI measurement times identified by considering T_m, T_DL, and T_CL were texture dependent, ranging from 45 min for silt to 120 min for silt loam and silty clay loam. For loamy sand, it was 50 min; for sand, it was 70 min, followed by 60 min for loam.     

太阳自动跟踪系统机构静力学及脉动风模拟分析

运用大型有限元软件ANSYS建立有限元模型,对太阳自动跟踪系统中的几个典型构件组合以及整体机构进行结构静力学模拟仿真分析,同时根据瞬态动力学理论模拟脉动风对机构的影响,探讨了模拟分析的细节问题并给出了求解全过程。组合结构以及整体机构的静力学模拟仿真分析结果表明了该机构可以达到设计要求。通过对机构进行脉动风共振模拟试验,由瞬态动力学分析结果表明脉动风使机构产生的振动频率小于固有频率,机构不会发生共振现象。并通过对两种模拟分析结果的比较,表明脉动风对机构的受力影响大。     

3-D Simulation on the Transient Heat Transfer Characteristics of Municipal Solid Waste Lump in Clean Incinerator

To be effectively heated in incinerator for MSW is very important for stabilization and completeness combustion, the smaller produce of the PCDD/Fs. At the same time, the transient heat transfer characteristics of the MSW lump decides the stay time of the MSW in incinerator and effects on the operation feature of the combustion equipments. All of these would lead to design the reasonable and clean combustion project, the optimum incinerator and recover energy system. The component, shape, dimension, property, the hardly and inner structure feature of the MSW lump are taken in account, and the effective conduct heat coefficient has been adopted. And then, the physical model is setup with different shape(ball, cylinder, squareness), and the 3-D mathematical model and conduct heat efficient with porous feature for transient heat transfer of MSW lump are presented, at the same time, the simulation tests are gotten in various boundary and the interesting conclusions are gotten: The center stable temperature of the MSW lump is mostly decided by the inner gas temperature of the incinerator and the heated area of the lump. The transient heat transfer characteristics is variously effected by the shape of the MSW lump in incinerator. The transient process becomes longer and the center stable temperature decreases, the volatile can't be easily separated out and the combustion efficiency decreases because of the increasing of the lump feature dimension.     

Analysis on Wind-induced Dynamic Response of Arched Corrugated Metal Roof

It is generally considered that the seismic response of arched corrugated metal roof is not the control factor of structure design because of its light dead weight, but the wind-induced dynamic response on the structure is significant, so more attention must be paid to it. First, the wind simulation of the structure by AR model is carried out. Then, the TRANS method in the ANSYS is used to calculate the dynamic responses in time domain to compare the results with those in frequency domain in order to verify the validity of the two methods.