Construction and identification of eukaryotic expression vector of bcl-2 siRNA

AIM: To construct eukaryotic expression vector of small interfering RNA(siRNA) specific to bcl-2 and investigate the effect of recombinant plasmid on suppressing bladder cancer cell growth.METHODS: siRNA of bcl-2 gene was designed according to the principle of RNAi-based medicine, and was converted into cDNA coding expression of small hairpin RNAs(shRNA) of siRNA. The cDNA was synthesized and inserted into plasmid pGenesil-1. The recombinant eukaryotic expression vectors of pGenesil-1545 and pGenesil-1555 were controlled by the U6 promoter of RNA polymerase Ⅲ, identified by the restriction map and the sequence analysis, and transfected into T24 cells. After T24 cells were transfected for 72 h, expression of bcl-2 mRNA was assayed by RT-PCR; and MTT was used to observe the proliferation of T24 cells.RESULTS: The recombinant plasmids of pGenesil-1545 and pGenesil-1555 were identified by the restriction map and the sequence analysis. The sequences completely coincided with the designs. The expression of the bcl-2 mRNA in T24 cells transfected with recombinant plasmid decreased nearly 80%, and the growth of T24 cells was suppressed significantly.CONCLUSION: The siRNA eukaryotic expression vector against bcl-2 gene is successfully constructed. It effectively downregulates the expression of bcl-2 in T24 cells and suppresses the cell growth.     

膜孔灌肥液自由入渗硝态氮运移特性试验研究

以KNO3为入渗溶液,模拟研究了硝态氮在膜孔灌肥液自由入渗条件下的运移过程.结果表明,在湿润蜂运移的范围内,沿着膜孔水平方向和垂直方向,硝态氮前锋运移速率受土壤含水量的影响较大,并随着土壤含水量的增加而增加;硝态氮前锋运移速率与运移距离有很好的相关性,并随着运移距离的增加呈幂函数衰减变化.硝态氮前锋浓度随着运移距离的增加而升高,其变化趋势呈指数曲线关系,且在湿润锋(土壤干湿交界)面上达到最大值;硝态氮前锋浓度随土壤含水量的增加呈幂函数递减,两者的相关系数也达到了极显著水平.     

沙漠地区风沙活动特征——以中国科学院风沙观测场为例

运用中国科学院风沙科学观测场的实测资料,对腾格里沙漠风沙环境特征进行初步分析,内容包括起沙风、风沙活动强度、风沙流结构和沙漠边界层的风速廓线.该地区的起沙风以6～8 m/s为主,占总起沙风的71.63%,其次为8～10 m/s,占19.24%,两者之和占90.87%;大风日数为4天;风向以W-N组风向为主;占全年的53.14%.年输沙势为36.56VU,风能属于低风能环境,单一主风向和单风态风环境.风沙流主要集中在地表的0.1 m,占总输沙量的95.46%.观测场近地层厚度大于50 m.     

AFLP analysis of Russian <Emphasis Type="Italic">Alternaria tenuissima</Emphasis> populations from wheat kernels and other hosts

Alternaria tenuissima is a common pathogen on a number of plants described in several geographic regions of the world. Genetic variation within and between Russian Far East, North West and Caucasus populations of A. tenuissima from wheat was examined. In addition, genetic differences between isolates from various hosts were estimated. In total, 101 isolates of A. tenuissima were studied using amplified fragment length polymorphism (AFLP) with four primer combinations. Wright’s fixation index (F _st), gene flow (N _m) and gene diversity (H _s) were calculated. AFLP banding patterns indicated significant genetic distance and at the same time a low level of gene flow between the Far East and the two other groups of isolates originating from the European part of country. The degree of similarity between the North West and Caucasus populations was very high, as was the migration rate. Isolates analysed by UPGMA-based cluster analysis were grouped according to location of origin but irrespective of plant host. Based on the F _st value, the group of isolates originating from wheat and barley were not found to differ significantly from each other.     

基于DSP与超声波测距的农业机器人定位与避障控制

随着新兴电子集成技术和自动化技术的发展,控制系统已逐渐向数字化转变,高集成芯片广泛应用于自动化控制领域,体积小、运算能力强的嵌入式系统慢慢开始取代计算机。为此,首先分析了超声波测距功能及其优越性,采用三球定位技术,设计研究了一种基于DSP和超声波的全局定位系统;然后,采用多路超声波收发模块设计了基于超声波的自主避障控制系统,并提出一种新的模糊推理方法,实现机器人的避障和路径规划功能;最后,采用Visual C++可视化程序设计软件对避障系统进行仿真和场地试验,验证了系统的可靠性和可行性,为机器人的研究和发展提供了更加宽广的空间。     

RNAi技术在转基因西瓜抗病毒病上的研究与应用

对RNAi技术的概念、研究进展及西瓜病毒病的危害及其发生特点、RNAi技术在西瓜抗病毒病上的研究进展进行了阐述,并对存在的一些问题作了简要探讨与展望。     

Comparative Study of Nutrient Deficiencies on Growth and Photochemistry of Tobacco

Abstract

Element deficiencies, such as molybdenum (Mo), iron (Fe), calcium (Ca), lead to disturbances of morphological and physiological processes of tobacco (Nicotiana tabacum L.). On exposure to nutrient solution without Mo, leaf morphology was significantly affected, whereas photosynthetic processes conserved their normal activities. The decreased Mo, Fe, and Ca concentrations in tobacco were associated with loss of pigments and photosynthetic disturbances. In Ca‐deficient plants a reduction in the rate constant of energy trapping by PSII centers and a physical dissociation of LHC from PSII core, were observed. The poisonous action of Fe and Ca deficiencies focused mainly in the decreased proportion of active chlorophyll associated with the reaction center (RC) of PSII (decreased F _v /F ₀) and in the declined total number of electrons that have gone through the RCs (decreased Area/F _m  ? F ₀). In parallel, the non‐photochemical quenching coefficient was significantly enhanced in Fe and Ca‐deficient plants, but remained unchanged in Mo‐deficient plants. It is obvious that nutrient deficiency may trigger some protective mechanisms in order to PSII could maintain its activity under that type of stress.     

靶向S基因外源性microRNA抑制TGEV增殖效果的研究

应用生物信息学软件对猪传染性胃肠炎病毒(TGEV)S基因进行分析,筛选出可能与S基因有相互作用的外源miRNA:amiRNA-S-28035,amiRNA-S-28038,amiRNA-S-28165。之后,利用脂质体将构建好的相应的表达载体瞬时转染至PK-15细胞;通过Q-PCR和间接免疫荧光方法检测其对S基因的抑制作用;CPE分析和TCID50测定检测其对TGEV增殖的抑制效果。结果发现,3个外源性miRNA均降低了S基因mRNA的转录和蛋白的表达,其中amiRNA-S-28038对TGEV mRNA的平均抑制率可达64.6%,最高可达69.9%,表明外源性microRNA可以通过靶向TGEV基因组来抑制TGEV的复制。研究结果为猪传染性胃肠炎的预防和治疗提供了新的思路。     

小麦小分子RNA TaMIR1129介导植株抵御低氮逆境功能研究

本研究针对迄今有关小麦小分子RNA(miRNA)家族成员介导植株氮素吸收和利用机理尚少见报道的现状,对TaMIR1129的表达特征和介导植株抵御低氮逆境功能进行了研究。结果表明,TaMIR1129呈低氮胁迫诱导表达,表现为随氮浓度降低(0.02~6mmol/L)和处理时间延长(0~48h)表达水平不断增高特征。此外,低氮诱导的高表达水平在恢复供氮后表达下调。表明该miRNA对介质中氮素应答呈典型的时间及浓度依赖特征。TaMIR1129作用2个靶基因,包括Molybdenum cofactor sulfurase(TaMCS)和Major facilitator family transporter(TaMFFT),上述基因应答低氮特征与TaMIR1129相反。遗传转化结果表明,超表达TaMIR1129具有显著增强植株抵御低氮逆境的能力。表现为与野生型对照相比,转基因系Sen 1和Sen 2低氮处理后植株形态增大,干质量增加,氮累积量增多。表明TaMIR1129与作用靶基因构建miRNA/target模块在介导植株抵御低氮逆境中发挥重要作用。     

运输对荣昌猪生理生化指标的影响

获取荣昌猪对运输的耐受能力,以此研究针对运输的保健药品和保健措施。选择40日龄的荣昌仔猪12头,在最低气温15℃下运输2h(100km)。从前腔静脉采集运输前及运输停止后30min、运输停止后第1天、3天、7天试验组及空白对照组荣昌仔猪的血液,进行主要血液生理生化指标检测。同时,检测试验组及空白对照组荣昌仔猪在运输前及运输停止后30min、运输停止后第1天、第3天、第7天的临床三大生理指标(呼吸、脉搏、体温)。采用F检验统计分析运输前后差异显著性,比较运输对组内各组影响的差异性。结果表明,荣昌仔猪在经过运输后,尿素、肌酐、总胆固醇、甘油三酯、谷丙转氨酶、谷草转氨酶、碱性磷酸酶等大部分生理生化指标都有上升,与运输前比较差异极显著(P<0.01),运输前各荣昌仔猪之间的生理生化指标无显著差异(P>0.05),荣昌猪对运输的抵抗能力较差,需要在运输前后做好抗应激措施。