大黄鱼肿瘤抑制因子cylindromatosis (LcCYLD)的序列与免疫反应特征及其功能

为研究大黄鱼肿瘤抑制因子cylindromatosis（CYLD）在免疫反应中的作用,本实验克隆了大黄鱼CYLD的全长cDNA（命名为LcCYLD）并对其进行序列分析;采用荧光定量PCR的方法对大黄鱼各组织及免疫刺激后的大黄鱼肾细胞系中的LcCY LD表达变化进行检测;构建了重组表达载体pTurboGFP-CYLD及pcDNA3.1-CYLD,分别用于亚细胞定位实验及过表达实验;在HEK293T细胞系中过表达LcCYLD后采用双荧光素酶报告系统检测了 NF-κB、TNF-α及IL-1β 启动子活性的变化。结果显示,LcCYLD的ORF包含2 754 bp,编码917个氨基酸,推测具有保守的3个N端的CAP-GLY结构域,1个磷酸化区域和1个C端的UCH结构域,多序列比对结果显示各物种CYLD间高度保守;系统进化分析显示,LcCYLD与来源于其他硬骨鱼的CYLD聚为一支,其中与条纹鲈的CYLD关系最近;转录水平表达分析发现LcCYLD在大黄鱼各组织均有表达,其中在脑中表达量最高;LPS及poly I:C刺激能够显著诱导LcCYLD的表达;亚细胞定位实验表明LcCYLD在细胞质及细胞核中均有表达;过表达LcCYLD能够显著抑制NF-κB及促炎细胞因子TNF-α及IL-1β的转录激活。以上研究结果表明大黄鱼CYLD能够抑制NF-κB的转录激活,为深入了解LcCYLD在大黄鱼先天免疫信号转导中的作用奠定基础。     

Characterization of cytoplasmic male sterility in Petunia hybrida and Zea mays. Localization and activity of cytochrome c oxidase

Summary Anthers of male fertile, cytoplasmic male sterile (cms), and restored male fertile Petunia hybrida, are analyzed for cytochrome c oxidase (cox) activity in subsequent stages of microsporogenesis, and compared with anthers of male fertile, cms-S and cms-C Zea mays. The cox activity is determined in anther extracts and cytochemically. In petunia anthers, the first differences in cox activity occur from meiosis onward. However, at these stages, the initial symptoms of degeneration are already apparent. It is suggested that the decline in enzyme activity of the cms petunia anthers is the result rather than the cause of the non-formation of functional pollen.In maize anthers, the cox activity of sterile-type anthers is reduced in comparison with fertile-type anthers from premeiosis onward. There are also consistent cytochemical differences in the mitochondrial organization of cox activity between pollen of cms-S and male ferile maize anthers. In fertile-type mitochondria, the DAB reaction product indicating cox activity is localized in the cristae and within the space between the outer and inner limiting membranes of the organelles. In mitochondria of pollen of cms-S maize, cox activity is only observed between the outer and inner membranes of the mitochondria. The biochemical and cytochemical differences are observed at stages of development at which no structural signs of degeneration are apparent. The results suggest that cms in maize correlates with deviations in cytochrome c oxidase activity.Abbreviations cox cytochrome c oxidase - DAB 3,3-diaminobenzidine     

Genetic Analysis of Chromosome 2D of Wheat

The Yugoslavian varieties ‘Novosadska Rana 1’ and ‘Sava’ are shown by monosomic comparisons to carry weak height promoters on chromosome 2D characteristic of the ‘Akakomugi’ gene for reduced height, Rht8. Reciprocal monosomic crosses between ‘Bersee’ and ‘Sava’ demonstrate ‘Sava’ chromosome 2D reduces height by about 16 cms, accelerates ear emergence by about 9 days and increases yield through increased grain number and grain size. Recombinant lines developed for chromosome 2D suggest that this chromosome in Mediterranean wheats carries three genes, Rht8, Ppd1 and Yr16, important to their adaptation. Rht8 and Ppd1, a gene for day length insensitivity together reduce height. Ppd1 and, to a minor degree, either yr16, the susceptible allele of a gene for adult plant resistance to yellow rust or a closely linked gene, accelerate time to flowering and thereby avoid desiccating Yugoslavian summer conditions. The same genes reduce spikelets numbers but this is offset by increased floret fertility producing an overall increase in the number of grains per ear. Ppd1 also by avoiding desiccating conditions increases gram size and together with either yr16 or the closely linked fertility gene increases ear and plant yields.     

Maternal and embryonal control of seed colour by different Brassica alboglabra chromosomes

Most oilseed rape, Brassica napus, cultivars are black‐seeded. The progenitor species, Brassica rapa, has either yellow or black seeds, while known cultivars of the other progenitor species Brassica oleracea/alboglabra have black seeds. To determine which chromosomes of B. alboglabra are carriers of seed colour genes, B. rapaalboglabra monosomic addition lines were produced from a B. napus resynthesized from yellow‐seeded B. rapa and brown/black‐seeded B. alboglabra. Eight out of nine possible lines have been developed and transmission frequencies of the alien chromosomes were estimated. Three B. alboglabra chromosomes in three of these lines influenced seed colour. B. rapa plants carrying alien chromosome 1 exhibited a maternal control of seed colour and produced only brown seeds, which gave rise to plants with either yellow or brown seeds. However, B. rapa plants carrying alien chromosome 4 or another as yet unidentified alien chromosome exhibited an embryonal control of seed colour and produced a mixture of yellow and brown seeds. The yellow seeds gave rise to yellow‐seeded plants, while the brown seeds gave rise to plants that yielded a mixture of yellow and brown seeds, depending on the absence or presence, respectively, of the B. alboglabra chromosome. Consequently, both maternal and embryonal control of seed colour are expected to contribute to the black‐seeded phenotype of oilseed rape.     

植物蛋白质的亚细胞定位研究进展

细胞是生命形式的基本组成单元,各种蛋白质按照其功能有序地分布在细胞的每个分区中。植物细胞的主要分区包括细胞膜和其他内膜系统、细胞核、细胞质以及位于其中的线粒体、叶绿体、高尔基体和内质网等各种细胞器。植物蛋白质的亚细胞定位是功能基因组学的重要内容。主要的植物蛋白质亚细胞定位技术包括:融合报告基因定位法、免疫组织化学定位法、蛋白质组学定位技术以及共分离标记酶辅助定位法,而生物信息学预测也成为亚细胞定位研究的一种重要手段。高通量蛋白质亚细胞定位技术的发展和应用,为构建定位数据库积累了数据。在模式植物拟南芥中,有亚细胞定位信息的蛋白质已经超过4 000个。     

银杏枝条营养贮藏蛋白质的免疫细胞化学鉴定

依据银杏枝条中32和36 ku蛋白质含量的明显季节变化,推测它们是银杏的营养贮藏蛋白质.采用间接免疫组织细胞化学定位技术,对这2种蛋白质进行细胞化学定位.32和36 ku蛋白质具有一定的免疫相关性,它们分布在一些皮层薄壁细胞和韧皮部的韧皮薄壁细胞中,存在于这些细胞的小液泡里.研究结果揭示这2种蛋白质的组织分布式样,为这2种蛋白质是银杏的营养贮藏蛋白质提供有力证据.     

基于乡土化的高校景观特色营造研究——山东交通学院长清校区景观设计解析

在对乡土化高校景观解读的基础上,分析其建设的迫切性,通过对山东交通学院长清校区景观设计的解析,探讨在高校景观设计中,如何分析利用基地条件和挖掘乡土素材进行设计,塑造具有乡土特色的校园环境。最后通过对中国已建成的比较优秀的乡土化高校景观的对比分析,提出进一步改进的对策。     

水稻红莲型粤泰不育花药ATPase超微结构定位

 以ATPase定位研究了红莲型粤泰不育花药和可育花药。结果表明 ,单核中位期可育花粉ATPase定位于覆盖层外侧、柱状层和细胞核 ,单核边位期ATPase定位于覆盖层外侧和细胞核 ;二核期ATPase定位于柱状层、内壁、质膜以及细胞核上 ;三核期ATPase定位于柱状层、内壁、质膜生殖核和营养核的核仁上。单核中位期不育花粉ATPase定位于核仁 ,单核边位期ATPase定位于覆盖层外侧、柱状层、质膜以及细胞核上 ;二核期不育花粉ATPase定位于覆盖层外侧、内壁和核仁。单核中位期可育花药药壁发育良好 ,绒毡层吞噬泡有活跃的ATPase活性 ,单核边位期绒毡层内吞噬泡消失 ,绒毡层内含物减少 ,二核期绒毡层细胞完全解体。单核中位期不育花药药壁发育不良 ,绒毡层内吞噬泡少 ,单核边位期绒毡层出现大量吞噬泡 ,二核期绒毡层存在尚未解体的细胞质和核。大多数不育花药药隔结构及ATPase定位特征与可育花药相似。可能不育花药绒毡层发育迟缓、单核边位期不育花粉细胞质膜活跃的ATPase活性及花粉核异常与花粉败育存在较为密切的关系。     

Advances in Localization and Molecular Markers of Wheat Leaf Rust Resistance Genes

Genetic resistance is the most economical method of reducing yield losses caused by wheat leaf rust. To identify the leaf rust resistance genes in commonly used parental germplasm and released cultivars become very important for utilizing the genetic resistance tc wheat leaf rust fully. Up to date, about 90 leaf rust resistance genes have been found,of which 51 genes have been located and mapped to special chromosomes, and 56 genes have been designated officially according to the standards set forth in the Catalogue of Gene Symbols for wheat. Twenty-four wheat leaf rust resistance genes have been developed for their molecular markers. It is very important to isolate, characterize, and map leaf rust resistance genes due to the resistance losses of the genes caused by the pathogen continuously.     

猪CFL2b/荧光蛋白融合基因在小鼠成肌细胞中的表达与定位

[目的]了解猪CFL2b基因在成肌细胞中的表达与定位。[方法]用RT-PCR方法扩增得到猪CFL2b基因,连接到T载体上,双酶切后通过定向克隆将其与真核表达载体pEGFP-N1的绿色荧光蛋白基因融合,构建CFL2b-荧光蛋白融合基因表达载体。然后经真核表达质粒-脂质体介导,导入C2C12细胞系。[结果]荧光显微镜观察显示:在空白载体pEGFP-N1转染的C2C12细胞中荧光布满整个细胞,而在转染阳性载体pEGFP-N1-CFL2b的C2C12细胞中荧光主要聚集在细胞质的肌动蛋白丝。表明转染的C2C12细胞系能够高效表达猪CFL2b蛋白,猪CFL2b基因表达的蛋白定位于细胞质中。[结论]该研究为CFL2b基因的功能及该基因与猪肉质性状的相关性的研究奠定了基础。