柴达木栽培和野生枸杞子中东莨菪内酯含量的测定北大核心CSCD

建立测定枸杞子中东莨菪内酯的HPLC方法,并进行方法学考察。利用所建立的方法对柴达木栽培和野生枸杞子的东莨菪内酯含量进行测定。以Diamonsil C18柱为分析柱,甲醇-0.05%磷酸溶液（35∶65）为流动相,流速：1 m L/min,柱温：30℃,检测波长：344 nm;研究发现,枸杞子中东莨菪内酯在0.026-1.040μg（R2=0.9996）范围内成良好的线性关系;平均加样回收率（n=6）为96.1%,RSD=2.0%。该方法准确、可靠、重现性好,适用于枸杞子中东莨菪内酯的含量测定。测定结果表明,柴达木栽培枸杞子具有很整齐的品质优良性,而野生枸杞子品质差异很大,适宜于选择性的开发。     

Expression of outer capsid protein VP5 of grass carp reovirus in <Emphasis Type="Italic">E.coli</Emphasis> and analysis of its immunogenicity

Grass carp reovirus (GCRV) is a tentative member of the Aquareovirus genus in the family Reoviridae. The mature virion comprises 11 dsRNA genomes enclosed by two concentric icosahedral proteins shells that is comprised of five core proteins and two outer capsid proteins. The genome sequence and 3D structure demonstrate there is a higher level of sequence homology in structural proteins between GCRV and mammalian orthoreoviruses (MRV) compared to other members of the family. To understand the pathogenesis of GCRV infection, the outer capsid protein VP5, a homology of the μ1 protein of MRV, was expressed in E.coli. It was found that the recombinant VP5 was highly expressed, and the expressed His-tag fusion protein was involved in the formation of the inclusion body. Additionally, specific anti-VP5 serum was prepared from purified protein and western blot demonstrated that the expressed protein was able to bind immunologically to rabbit anti GCRV particle serum and the immunogenicity was determined by ELISA assay. Additional experiments in investigating the functional properties of VP5 will further elucidate the role of the GCRV outer capsid protein VP5 during entry into host cells, and its interaction among viral proteins and host cells during the infection process.     

Expression and identification of inclusion forming-related domain of NS80 nonstructural protein of grass carp reovirus

Grass carp reovirus (GCRV), a double stranded RNA virus that infects aquatic animals, often with disastrous effects, belongs to the genus Aquareovirus and family Reoviridea. Similar to other reoviruses, genome replication of GCRV in infected cells occurs in cytoplasmic inclusion bodies, also called viral factories. Sequences analysis revealed the nonstructural protein NS80, encoded by GCRV segment 4, has a high similarity with uNS in MRV(Mammalian orthoreoviruses), which may be associated with viral factory formation. To understand the function of the uNS80 protein in virus replication, the initial expression and identification of the immunogenicity of the GCRV NS80 protein inclusion forming-related region (335.742) was investigated in this study. It is shown that the over-expressed fusion protein was produced by inducing with IPTG at 28oC. In addition, serum specific rabbit antibody was obtained by using super purified recombinant NS80(335.742) protein as antigen. Moreover, the expressed protein was able to bind to anti-his-tag monoclonal antibody (mouse) and NS80(335-742) specific rabbit antibody. Further western blot analysis indicates that the antiserum could detect NS80 or NS80C protein expression in GCRV infected cells. This data provides a foundation for further investigation of the role of NS80 in viral inclusion formation and virion assembly.     

黑曲霉β-葡萄糖苷酶An-bgl3的重组表达及东莨菪苷的转化

东莨菪素是一种香豆素类化合物,有消肿抗炎、镇痛、抗菌、杀螨等生物活性。从植物中提取东莨菪素由于东莨菪苷及其他成分的干扰常导致提取率低、纯化困难。本研究对黑曲霉(Aspergillus niger)来源的β-葡萄糖苷酶基因An-bgl3进行异源表达、纯化及表征,分析了该酶与东莨菪苷的构效关系,并研究其对植物粗提物中东莨菪苷的转化作用。结果表明,纯化后β-葡萄糖苷酶An-bgl3的比活力为15.22 IU/mg;表观分子量约为120 kDa;最适反应温度和pH分别为55℃和4.0。10 mmol/L的金属离子Fe²⁺和Mn²⁺可使该酶活力活分别提高1.74倍和1.20倍;10 mmol/L的Tween-20、Tween-80和Triton X-100对于该酶的抑制效果均为30%;能够耐受10%浓度的甲醇和乙醇溶剂。东莨菪苷与An-bgl3具有较好的亲和作用。该酶处理丁公藤(Erycibe obtusifolia Benth)粗提物,能够专一性地将东莨菪苷水解为东莨菪素,使东莨菪素含量增加47.8%。本研究表明黑曲霉β-葡萄糖苷酶An-bgl3对东莨菪苷具有专一性,且酶学性质良好。该研究为使用β-葡萄糖苷酶资源从植物材料中提取东莨菪素提供了参考。     

东濮地区奥陶纪牙形石

本文记叙了东濮地区井下奥陶纪牙形石１６属３２种（１新种、３未定种、４未定属种）,识别出５个牙形石带,１个未建带,自下而上为１．Ｓｃｏｌｏｐｏｄｕｓｆｌｅｘｉｌｉｓ带;２．Ｔａｎｇｓｈａｎｏｄｕｓｔａｎｇｓｈａｎｅｎｓｉｓ带;３．未建带;４．Ｐｌｅｃｔｏｄｉｎａｏｎｙｃｈｏｄｏｎｔａ带;５．Ａｕｒｉｌｏｂｏｄｕｓｓｅｒｒａｔｕｓ带;６．Ｍｉｃｒｏｃｏｅｌｏｄｕｓａｓｙｍｍｅｔｒｉｃｕｓ－Ｂｅｌｏｄｉｎａｃｏｍｐｒｅｓｓａ带。     

东太湖全新世水下河道沉积特征及环境演变

在详细研究东太湖Ｅ２孔全新世沉积基础上,总结和归纳了水下河道的沉积特征和相组合规律。识别出高湖面型和低湖南型两种沉积层序,划分了东太湖晚更新世末以来环境演变阶段：１）１５８８５－６５７５ａＢ．Ｐ．冲积黄土基础上发育而成的古河道;２）６５７５－５８００ａＢ．Ｐ．高湖面时期水下河道沉积环境;３）５８００ａＢ．Ｐ．后,演化为低湖面型水下河道,并逐渐淤积形成开敞湖缘环境。结合西太湖Ｗ１孔沉积特征和层位对比     

广东滩涂经济鱼类寄生吸虫Ⅱ．异肉科和独睾科吸虫──（吸虫纲：复殖目）

本文对广东滩涂经济鱼类寄生吸虫异肉科Ａｌｌｏｃｒｅａｄｉｉｄａｅ（Ｌｏｏｓｅ,１９０２）（Ｓｔｏｓｓｉｃｈ,１９０３）和独睾科ＭｏｎｏｒｃｈｉｉｄａｅＯｄｈｎｅｒ,（１９１１）进行研究,并建立拟履口吸虫属Ｐａｒａｃｒｅｐｉｄｏｓｔｏｍｕｍｇｅｎ．ｎｏｖ．是异肉科,履口亚科Ｃｒｅｐｉｄｏｓｔｏｍｉｎａｅ（Ｄｏｌｌｆｕｓ,１９５１）的新属;长宫吸虫属Ｌｅｎｏｌｏｐａｇｅｎ．ｎｏｖ是独睾科、孤睾亚科Ｌａｓｉｏｔｏｃｉｎａｅ的新属。文中对二新属二新种：金钱鱼拟履口吸虫Ｐａｒａｃｒｅｐｉｄｏｓｔｏｍｕｍｓｃａｔｏｐｈａｇｕｓｉ和惠州长空吸虫Ｌｅｎｏｌｏｐａｈｕｉｚｈｏｕｅｎｓｉｓ分别进行了详细的描述,并对异肉科、履口亚科和属的分类特征进行修正、补充和讨论。     

东祁连山退化高寒草地土壤细菌群落与土壤环境因子间的相互关系

为探究东祁连山不同退化高寒草地细菌群落分布特征与土壤环境因子间的相互关系,采用高通量测序技术对轻度、中度和重度退化草地的土壤细菌群落结构变化及其多样性进行分析,运用CANOCO 4.5软件对土壤细菌群落与土壤环境因子间关系进行冗余分析(RDA).结果表明:不同退化高寒草地土壤理化性质间均差异显著,高通量测序共得到257125条有效序列,180826条优质序列,4790个OTUs.细菌群落Chao1指数依次为轻度>中度>重度;Shannon指数依次为轻度>重度>中度.系统发育分析表明,各样地土壤细菌类群分属于33个门,其中放线菌门、变形菌门和厚壁菌门是3种不同退化草地土壤中的优势类群.对不同退化草地土壤细菌各门所占比例分析发现,放线菌门、酸杆菌门和变形菌门随着退化程度加剧先减少后增加,厚壁菌门反之.RDA分析结果显示,细菌优势类群与蔗糖酶、纤维素酶和磷酸酶呈极显著相关,与pH、电导率、速效氮、速效钾呈显著相关.说明东祁连山不同退化高寒草地土壤细菌群落间差异明显,土壤环境因子是影响土壤细菌群落分布的重要因素.     

东太平洋中部中上层鲨鱼群落营养生态位分化

鲨鱼在大洋生态系统中占据着重要的生态地位,其作为顶级捕食者,通过下行效应直接影响生态系统的稳定.稳定同位素技术是目前研究摄食生态学强有力的手段之一,可利用碳氮稳定同位素在食物网中的特性分别指示鲨鱼的食物来源和营养级.本研究选取8种130尾采集自东太平洋中部的中上层鲨鱼,应用稳定同位素绘制其种群生态位图谱,比较不同种群间的生态地位及资源分配方式上的差异.结果表明:不同鲨鱼种群碳、氮稳定同位素比值存在显著差异;8种鲨鱼在东太平洋生态系统中的营养级为4.3~5.4,大青鲨、尖吻鲭鲨与其他6种鲨鱼存在摄食隔离,表现出独特的营养生态地位.这些结果充分证明大洋性中上层鲨鱼并非生态系统的冗余种,其营养生态位的独特性不会被其他捕食者简单地替代和弥补.