四川省中药资源特征与开发利用研究

四川省中药资源具有种类多,蕴藏量大,水平分布和垂直分布明显,地道药材质量好,民族药物丰富和栽培药材产值高等特点。1986年统计,全省有中草药4550种。其中,植物药3962种,动物药344种,矿物药44种,产量和产值均约占全国的1/10。四川省在开发利用中药中,存在着资源蕴藏量不清、常用中药减少、产销失调、加工能力低等问题。建议在中药开发利用机构、决策系统、资源政策、生产经营和保护方面采取措施,合理利用中药资源。     

我国土壤中苏云金芽孢杆菌的分离与基因型的鉴定

从我国20个省市采集176份土壤中分离出苏云金芽孢杆菌51株.以cry1,cry2,cry3,cry4,cry-n,cry7,cry8,cry11,cry13和cyt的引物作PCR分析.结果表明:4株含有cry4,12株含有cry7,4株含有cry8,4株含有cry1,2株同时含有cry1和cry2,27株无PCR产物.未检测到其它基因型.经电镜扫描伴胞晶体形态多种多样,主要有菱形、球形、方形、多边形和不规则形.图3表3参15     

水稻花药绒毡层特异表达启动子的分离与表达载体构建

从籼稻川75A(OryzasativaL.)黄化苗叶片中直接提取基因组总DNA.根据文献报道的水稻花药绒毡层特异启动子Osg6B序列设计引物,采用TouchdownPCR技术获得水稻花药绒毡层特异表达的启动子Tsp1,将该片段克隆在载体pMD18-TVector上转化到大肠杆菌JM109,并酶切和PCR验证.序列分析发现,该片段与Osg6B启动子同源性为96%,且具有真核生物启动子的多种特征序列,表明启动子Tsp1为来源于籼稻川75A中的水稻花药绒毡层特异表达的启动子.利用该启动子对本实验室保存的质粒pIB467和pIB009进行改造,构建了一个可产生新型雄性不育系的双元植物表达载体pJH401.图7参13     

化学-生物絮凝污水处理工艺中微生物群落结构变化分析

利用分子生物学技术，直接从化学-生物絮凝工艺的活性污泥样品中提取DNA，对16S rDNA V3区进行PCR扩增，结合DGGE（变性浓度梯度凝胶电泳），分析了活性污泥中微生物群落结构，并对Shannon多样性指数进行分析讨论，通过研究指出系统中细菌数量的增加或减少．测定了活性污泥中部分菌种的16S rDNA V3区片段序列，通过NCBI（美国国立生物技术信息中心）基因库比对，初步确定细菌的属．结果表明，PCR—DGGE结合测序技术是一种完全可行的快速进行环境样品微生物研究的分析方法．图3表4参13     

稀有鮈鲫Dmrt基因家族13个成员的克隆与序列分析

已经发现果蝇Doublesex、线虫Mab-3、青DMRT1Y/DMY和人类DMRT1等性别决定与分化基因均含有一个具有DNA结合能力的保守基序--DM结构域,对性别决定和性别分化具有调控功能.利用简并PCR,从稀有鲫基因组DNA中克隆了13个具有不同DM结构域的Dmrt基因家族成员.基于DM保守基序,建立了各物种的进化树.结果表明,稀有鲫基因组存在多个Dmrt基因成员,该基因在脊椎动物和非脊椎动物中具有高度保守性,是一种理想的环境内分泌干扰物研究的分子模型,在分子生态毒理学研究中具有很好的应用前景.     

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     

Evaluating Effects of Habitat Loss and Land-Use Continuity on Ant Species Richness in Seminatural Grassland Remnants

Abstract:  Seminatural grasslands in Europe are susceptible to habitat destruction and fragmentation that result in negative effects on biodiversity because of increased isolation and area effects on extinction rate. However, even small habitat patches of seminatural grasslands might be of value for conservation and restoration of species richness in a landscape with a long history of management, which has been argued to lead to high species richness. We tested whether ant communities have been negatively affected by habitat loss and increased isolation of seminatural grasslands during the twentieth century. We examined species richness and community composition in seminatural grasslands of different size in a mosaic landscape in Central Sweden. Grasslands managed continuously over centuries harbored species-rich and ecologically diverse ant communities. Grassland remnant size had no effect on ant species richness. Small grassland remnants did not harbor a nested subset of the ant species of larger habitats. Community composition of ants was mainly affected by habitat conditions. Our results suggest that the abandonment of traditional land use and the encroachment of trees, rather than the effects of fragmentation, are important for species composition in seminatural grasslands. Our results highlight the importance of considering land-use continuity and dispersal ability of the focal organisms when examining the effects of habitat loss and fragmentation on biodiversity. Landscape history should be considered in conservation programs focusing on effects of land-use change.     

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