杂交水稻及其“三系”线粒体DNA的AP—PCR指纹图谱

为了研究水稻（Ｏｒｙｚａ ｓａｔｉｖａ Ｌ．）细胞质雄性不育（ＣＭＳ）与线粒体基因组的关系,应用ＡＰ－ＰＣＲ 分析,用７ 个任意单引物对６ 种水稻品系线粒体ＤＮＡ 进行了扩增。水稻线粒体ＤＮＡ 的ＡＰ－ＰＣＲ 产物可分为三种类型：（１）所有供试品系均能扩增的片段,它们代表了线粒体ＤＮＡ 在进化上的保守性序列。有４ 个引物检测到这类片段。（２）２ 个以上水稻品系共同出现而在全部供试材料间存在差异的扩增片段,这类片段是检测水稻线粒体ＤＮＡ多态性的主要来源。（３）一种细胞质类型所特有的扩增片段,从引物Ｒ２ 和Ｖ５ 的扩增产物中发现了这类片段,它们可能与ＣＭＳ有关联。另外,ＷＡ型不育系珍汕９７Ａ 与其杂种之间在６ 个引物的扩增图谱上均存在不同程度的差异,说明两者的线粒体ＤＮＡ序列结构可能存在某种差别     

Surface Display of Human Serum Albumin on Bacillus subtilis Spores for Oral Administration

Human serum albumin (HSA) is the major protein component of human plasma. To date, HSA for clinical uses is mostly produced by fractionation of human whole blood, which is accompanied by a lot of limitations. To obtain long-term bioactive albumin, we used hsa as a foreign gene and constructed a recombinant plasmid pJS700-HSA which carries a recombinant gene cotC-hsa under the control of cotC promoter. Plasmid pJS700-HSA was transformed into Bacillus subtilis by double cross-over and an amylase inactivated mutant was produced. After induction of spore formation, western blot and fluorescence immunoassay were used to monitor HSA surface expression on spores. We estimated that HSA displayed on the spore accounted for 0.135 % of the total spore proteins and about 0.023 fg HSA were exposed on the surface of each spore. Oral administration to mice with spores displaying HSA implied that the recombinant spores may have potential ability to increase the serum albumin level in vivo due to the resistant characters of spores.     

九龙江河口浮游植物的时空变动及主要影响因素

于2009年春（5月）、夏（8月）、秋（11月）在九龙江河口水域进行了水文、化学和生物的生态完全示范区综合外业调查,研究了九龙江河口浮游植物的种类组成、密度分布、季节变化、空间差异及主要影响因素,并结合前期资料分析了年际变动。结果表明,九龙江河口的浮游植物共记录7个门类75属134种。主体是硅藻,绿藻次之,甲藻和蓝藻较少,黄藻检出率高,裸藻和金藻零星检出。种类组成的空间差异大,绿藻在河口内区淡水水域比硅藻更占优势, 中肋骨条藻（Skeletonema costatum）、短角弯角藻（Eucampia zodiacus）、圆筛藻（Coscinodiscus spp.）、颗粒直链藻（Melosira granulata）、微小小环藻（Cyclotella. caspia）是河口区咸淡水水域及近海区的主要种类。浮球藻（Planktosphneria gelotinosa）、栅藻（Scenedesmus spp.）、盘星藻（Pediastrim spp.）、小席藻（Phormidium tenus）是河口内区淡水水域的主要种类。根据浮游植物的生态类型及其生境特征大致可分为三大类群。浮游植物密度夏季最高,平均为358.68103cells/L,密集中心的季节变化明显,密度分布由优势类群的密度分布决定。中肋骨条藻和短角弯角藻的数量庞大,导致优势种突出,多样性降低,种间分布不均匀,群落结构简单化。与史料比对,种类组成因淡水藻类的列入而更丰富,密度年际降低,中肋骨条藻仍是第一优势种,但优势度有较大降幅,优势类群有重大年际变化,细胞个体较小的种类占优。盐度和营养盐对浮游植物的分布及密度变化造成极大的时空差异,存在线性、复合线性、多项回归等复杂的相关关系。     

马铃薯StSnRK2.1基因克隆与生物信息学分析

SnRK2基因对植物的逆境胁迫具有重要的调节作用,以马铃薯‘陇薯3号’(Solanum tuberosum)为试材,采用RT-PCR方法从马铃薯试管苗中克隆得到1个SnRK2.1基因cDNA,命名为StSnRK2.1,提交GenBank注册,注册号为JX280911。通过生物信息学分析,该基因开放阅读框全长1 008 bp,编码335个氨基酸。预测蛋白质分子量约为37.77 kD,等电点为5.37,蛋白质二级结构预测α-螺旋42.39%,延伸链16.42%,β-折叠7.46%,无规卷曲33.73%,具有疏水性,为膜内蛋白。亚细胞定位显示该基因出现在细胞质及微体中的可能性较大。肽链可能有7处丝氨酸磷酸化位点,2处苏氨酸磷酸化位点,以及3处酪氨酸磷酸化位点,因此推测该基因在植物抗逆中有重要的作用。     

GS3, a major QTL for grain length and weight and minor QTL for grain width and thickness in rice, encodes a putative transmembrane protein

The GS3 locus located in the pericentromeric region of rice chromosome 3 has been frequently identified as a major QTL for both grain weight (a yield trait) and grain length (a quality trait) in the literature. Near isogenic lines of GS3 were developed by successive crossing and backcrossing Minghui 63 (large grain) with Chuan 7 (small grain), using Minghui 63 as the recurrent parent. Analysis of a random subpopulation of 201 individuals from the BC₃F₂ progeny confirmed that the GS3 locus explained 80–90% of the variation for grain weight and length in this population. In addition, this locus was resolved as a minor QTL for grain width and thickness. Using 1,384 individuals with recessive phenotype (large grain) from a total of 5,740 BC₃F₂ plants and 11 molecular markers based on sequence information, GS3 was mapped to a DNA fragment approximately 7.9 kb in length. A full-length cDNA corresponding to the target region was identified, which provided complete sequence information for the GS3 candidate. This gene consists of five exons and encodes 232 amino acids with a putative PEBP-like domain, a transmembrane region, a putative TNFR/NGFR family cysteine-rich domain and a VWFC module. Comparative sequencing analysis identified a nonsense mutation, shared among all the large-grain varieties tested in comparison with the small grain varieties, in the second exon of the putative GS3 gene. This mutation causes a 178-aa truncation in the C-terminus of the predicted protein, suggesting that GS3 may function as a negative regulator for grain size. Cloning of such a gene provided the opportunity for fully characterizing the regulatory mechanism and related processes during grain development.     

Large-scale recording of neurons by movable silicon probes in behaving rodents

A major challenge in neuroscience is linking behavior to the collective activity of neural assemblies. Understanding of input-output relationships of neurons and circuits requires methods with the spatial selectivity and temporal resolution appropriate for mechanistic analysis of neural ensembles in the behaving animal, i.e. recording of representatively large samples of isolated single neurons. Ensemble monitoring of neuronal activity has progressed remarkably in the past decade in both small and large-brained animals, including human subjects. Multiple-site recording with silicon-based devices are particularly effective because of their scalability, small volume and geometric design. Here, we describe methods for recording multiple single neurons and local field potential in behaving rodents, using commercially available micro-machined silicon probes with custom-made accessory components. There are two basic options for interfacing silicon probes to preamplifiers: printed circuit boards and flexible cables. Probe supplying companies (http://www.neuronexustech.com/; http://www.sbmicrosystems.com/; http://www.acreo.se/) usually provide the bonding service and deliver probes bonded to printed circuit boards or flexible cables. Here, we describe the implantation of a 4-shank, 32-site probe attached to flexible polyimide cable, and mounted on a movable microdrive. Each step of the probe preparation, microdrive construction and surgery is illustrated so that the end user can easily replicate the process.     

杏仁核点燃模型癫痫样放电传播途径研究

目的 :探讨杏仁核点燃模型癫痫样放电的传播途径。方法 :选择健康Wistar大鼠 3 0只以电刺激杏仁核的方式制作杏仁核点燃癫痫模型 ,于右侧杏仁核、左侧海马及右侧额叶皮质埋植电极记录脑电活动 ,观察电刺激杏仁核时在杏仁核、海马及额叶皮质出现癫痫样放电的潜伏期、最低刺激强度及癫痫样放电的持续时间。结果 :杏仁核出现癫痫样放电时 ,海马及皮质均未记录到癫痫样放电。而当杏仁核、海马及皮质三处出现癫痫样放电时的最低刺激强度依次增大 ,潜伏期依次延长 ,海马处癫痫样放电的持续时间最长。结论 :杏仁核点燃模型癫痫样放电可能由杏仁核经海马传至皮层 ,海马可能为癫痫样放电传播的重要结构     

九龙江河口浮游动物的群落结构和时空变动

2009年春(5月)、夏(8月)、秋(11月)在九龙江河口进行浮游动物采样调查,分析了九龙江河口浮游动物的种类组成、群落结构、丰度和生物量的时空变化。调查共发现浮游动物119种,其中,含桡足类(35种)和水母类(31种)是最主要的类群。主要优势种有火腿许水蚤(Schmackeria poplesia)、中华假磷虾(Pseudeuphausia sinica)、弗洲指突水母(Blackfordia virginica)、短尾类蚤状幼虫 (Zoea larvae)和长尾类幼虫 (Macrura larva)。其中,春季以火腿许水蚤、弗洲指突水母、中华假磷虾以主;夏季则以短尾类蚤状幼虫和长尾类幼虫为主;秋季则以火腿许水蚤和中华异水蚤(Acaritella sinensis)为主,季节变化明显。浮游动物丰度和生物量春季最高,平均值分别为3 377.8 ind/m³和3 209.4 mg/m³,秋季最低,但空间分布不均,从春季到秋季其密集中心从近海区逐渐转移到邻近河口区。与过去同期的调查资料相比,20年来九龙江河口浮游动物总丰度、优势种的组成和排序发生了显著的变化。本次浮游动物的总丰度平均值(以秋季为例)为历次调查最低,主要优势种也明显不同, 推测,浮游动物有朝着小型化发展的趋势。     

NOX4-Derived Reactive Oxygen Species Drive Apelin-13-Induced Vascular Smooth Muscle Cell Proliferation via the ERK Pathway

Apelin is the endogenous ligand of the G-protein-coupled receptor, apelin–angiotensin receptor-like 1 (APJ). Vascular smooth muscle cells express both apelin and APJ, which are important regulatory factors in the cardiovascular system. Apelin-13 significantly stimulated vascular smooth muscle cell proliferation. However, little is known about the precise cellular mechanisms responsible for vascular smooth muscle cell proliferation induced by apelin-13. Here, we present novel data that indicate the key role of NADPH oxidase 4-derived reactive oxygen species in proliferation of vascular smooth muscle cells treated with apelin-13. Apelin-13 stimulated reactive oxygen species production in a concentration- and time-dependent manner. Furthermore, DPI impaired apelin-13-induced reactive oxygen species generation and vascular smooth muscle cell proliferation. Apelin-13-treatment increased the expression of NADPH oxidase 4 in a dose-dependent manner. Down-regulation of NADPH oxidase 4 using siRNA prevented apelin-13-induced reactive oxygen species generation and vascular smooth muscle cell proliferation. An increase in reactive molecules can trigger the activation of ERK stress-sensitive signaling pathways. Additionally, siRNA-NOX4 and DPI reversed the phosphorylation of ERK induced by apelin-13. Apelin-13 induced vascular smooth muscle cell proliferation by NOX4-derived ROS via the ERK signaling pathway.