基于分子信标探针的荧光定量PCR方法检测转基因食品

选择了内源基因大豆植物凝集素(lectin)、玉米转化酶(invertase)和外源基因花椰菜花叶病毒35S(CaMV35S)启动子的分子信标探针,确定了探针浓度和镁离子浓度等反应条件,分别对转基因大豆和转基因玉米系列标准品进行内源基因和外源基因的荧光PCR扩增,在PCR反应过程中分别以两种荧光通道信号分别追踪同一样品DNA内源基因和外源基因的扩增动力学变化,并依此绘制了循环阈值与转基因食品百分比含量之间的标准曲线,建立了转基因大豆和转基因玉米的分子信标探针-荧光定量PCR检测方法,该方法具有特异性强、敏感性高的特点,实现了对转基因食品的定量分析.图7表1参11     

Analysis of parental strain DNA fragments existing in GEMs-Fhhh

There were 6 target DNA fragments of the three parental strains existing in the cell of GEMs( genetically engineered microorganism strain) Fhhh measured in this research by PCR(polymerase chain reaction). The determination showed that GEMs Fhhh contained all the 6 target DNA fragments, mnpl, mnp2, lipl, lip2, FLOI and 16S rDNA, and had the molecular genetic stability. Meanwhile the PCR production of each parental strain could only had its target DNA fragments and was different from each other. It may illustrate that the technique of the inter-kingdom protoplast fusion for the construction of GEMs Fhhh through the process of intercellular gene recombination could be used as a reliable bioengineefing technique to create the soecific functional stain for the nollution control.     

Isolation of fetal cells from transcervical samples by micromanipulation: Molecular confirmation of their fetal origin and diagnosis of fetal aneuploidy

Transcervical cell (TCC) samples have been shown to contain fetal cells amenable to molecular analysis. However, the presence of ‘contaminating’ maternal cells limits their use for prenatal diagnoses. In this report we show that clumps of fetal cells can be isolated from transcervical samples by micromanipulation and tested by fluorescence in situ hybridization (FISH) and polymerase chain reaction (PCR). Out of 129 clumps, isolated from mucus aspirates and transcervical lavages from 29 patients, 29 clumps from 11 patients were found to be exclusively of fetal origin as judged by the detection of chromosome 21-specific polymorphic DNA markers and Y-derived DNA sequences by PCR and FISH. One case of a male triploid fetus, diagnosed by the analysis of TCC samples obtained by mucus aspiration and lavage, was confirmed by testing clumps of cells isolated by micromanipulation.     

PCR-RFLP技术在环境地球化学研究中的应用及展望

PCR—RFLP技术为环境地球化学提供了一种新的研究方法和实验设计的新的思维方式。该方法具有所需样品量低、快速简便及特征性强等优点 ,可广泛应用于物质的生物地球化学循环、环境过程的生物作用、生物多样性及有机物源判断等方面的研究。利用PCR—RFLP技术 ,以环境中存在的 16SrRNA为对象 ,对环境生物的研究已广泛开展并取得了许多成果。展望未来研究成果 ,将会在海洋及湖泊沉积物等自然环境中发现更多的、新的微生物种类 ;将进一步阐明生物作用和物质循环的机理和过程 ;进一步阐明自然环境中生物大分子的变化机理及其环境效应并找到判断沉积物有机物源的新方法     

基于Ⅰ-Ⅱ复合型裂纹破坏模式的条带矿柱破坏宽度研究

采用条带法进行地下开采时,条带矿柱的裂纹扩展导致了矿柱的最终破坏,且裂纹多表现为Ⅰ型裂纹与Ⅱ型裂纹复合扩展破坏特征。根据采场矿柱受力条件,确定条带矿柱受两侧采场空区影响时矿柱内Ⅰ-П复合型裂纹尖端的Westergaard应力函数,采用HoekBrown强度准则计算采场作业面围岩破坏区的边界条件,建立了矿柱破坏宽度计算模型。以上横山页岩矿床为研究对象,采用物理相似模拟试验,模拟条带法回采某盘区3个试验区段,利用DIP法分析受地应力及回采扰动作用下矿柱的裂纹发育特征,验证模型的有效性。结果表明,条带及其条带矿柱均为15 m等设计参数条件下,计算模型矿柱破坏宽度理论值分别为17.66 m、15.56 m和10.28 m,试验模拟结果分别为完全大于15 m、约为15 m和10.5~11.0 m,试验结果与依据模型计算的理论值相近。     

青海省井孔水温观测结果分析——以德令哈等5个观测井为例

选取青海前兆台网的德令哈等5个井孔开展水温对比观测,对这五个井孔的水温原始观测数据和对比观测数据进行对比分析,从两组数据变化形态、差值、日变幅等方面发现不同的井孔两套数据的相似程度不同,其中以德令哈台为最好,湟源和格尔木次之,互助和佐署井则较差。分析认为主要原因有探头安装情况、仪器型号和仪器工作状态的影响。     

Repeated batch and continuous degradation of chlorpyrifos by Pseudomonas putida

The present study was undertaken with the objective of studying repeated batch and continuous degradation of chlorpyrifos (O,O-diethyl O-3,5,6-trichloropyridin-2-yl phosphorothioate) using Ca-alginate immobilized cells of Pseudomonas putida isolated from an agricultural soil, and to study the genes and enzymes involved in degradation. The study was carried out to reduce the toxicity of chlorpyrifos by degrading it to less toxic metabolites. Long-term stability of pesticide degradation was studied during repeated batch degradation of chlorpyrifos, which was carried out over a period of 50 days. Immobilized cells were able to show 65% degradation of chlorpyrifos at the end of the 50th cycle with a cell leakage of 112 × 10³ cfu mL^?1. During continuous treatment, 100% degradation was observed at 100 mL h^?1 flow rate with 2% chlorpyrifos, and with 10% concentration of chlorpyrifos 98% and 80% degradation was recorded at 20 mL h^?1 and 100 mL h^?1 flow rate respectively. The products of degradation detected by liquid chromatography–mass spectrometry analysis were 3,5,6-trichloro-2-pyridinol and chlorpyrifos oxon. Plasmid curing experiments with ethidium bromide indicated that genes responsible for the degradation of chlorpyrifos are present on the chromosome and not on the plasmid. The results of Polymerase chain reaction indicate that a ~890-bp product expected for mpd gene was present in Ps. putida. Enzymatic degradation studies indicated that the enzymes involved in the degradation of chlorpyrifos are membrane-bound. The study indicates that immobilized cells of Ps. putida have the potential to be used in bioremediation of water contaminated with chlorpyrifos.     

Enhancement of conservation knowledge through increased access to botanical information

Herbarium specimens are increasingly recognized as an important resource for conservation science and virtual herbaria are making specimens freely available to a wider range of users than ever before. Few virtual herbaria are designed with conservation use as a primary driver. Exceptionally, Brazil's Reflora Virtual Herbarium (RVH) was created to increase knowledge and conservation of the Brazilian flora. The RVH is closely integrated with the Flora of Brazil 2020 platform on which Brazil's new national Flora is under construction. Both resources are accessible via the Reflora home page and thousands of users move seamlessly between these Reflora resources. To understand how the Reflora resources are currently used and their impact on conservation science, we conducted a literature review and an online survey. We searched for publications of studies in which Reflora resources were used and publications resulting from Brazilian researchers who were part of Reflora's research and mobility program. The survey contained multiple choice questions and questions that required a written response. We targeted Reflora webpage visitors with the survey to capture a wider range of Reflora users than the literature review. Reflora resources were used for a variety of conservation-relevant purposes. Half the 806 scientific publications in which Reflora was cited and 81% of the 1069 survey respondents accessing Reflora resources mentioned conservation-relevant research outputs. Most conservation-relevant uses of the Reflora resources in scientific publications were research rather than implementation focused. The survey of Reflora users showed conservation uses and impacts of virtual herbaria were more numerous and diverse than the uses captured in the literature review. Virtual herbaria are vital resources for conservation science, but they must document use and impacts more comprehensively to ensure sustainability.     

化工园区安全生产监管存在问题及对策

系统分析了我国现阶段化工园区的基本特点及其安全监管中存在的问题，提出了建立以政府为主导、社会中介机构技术支撑、企业落实一体化的化工园区综合管理体系。实现化：亡园区应急救援资源整合及资源共享，提高化工园区应急救援资源有效利用率。数字化管理对化工园区的整体安全性的提高将发挥巨大的作用。