基因芯片技术在植物研究中的应用

简述了基因芯片的原理、特点、分类及制作方法,同时详细综述了基因芯片技术在植物研究中的应用。     

基因芯片技术及其应用研究

基因芯片技术是建立在杂交序列基本理论上的分子生物学技术,具有高度平行性、多样性、微型性和自动化的特点。由于其克服了传统核酸印迹杂交技术操作复杂、自动化程度低、检测效率低等缺点得以飞速发展,目前,该技术已用于DNA测序、基因表达分析、新基因的发现、基因单核苷酸多态性(SNPs)研究、基因诊断、药物筛选等领域。     

基因芯片技术及其在植物上的应用

基因芯片技术（gene chip technology)是采用光导原位合成或缩微印刷等方法,将大量特定的DNA探针片段有序地固定于固相载体的表面,形成DNA微阵列,然后与待测的标记样品靶DNA或RNA分子杂交,对杂交信号进行扫描及计算机检测分析,从而获取所需的生物信息。该技术在植物研究中广泛应用于寻找特异性相关基因和新基因,基因表达分析,基因突变和多态性检测,DNA测序等。     

基因芯片在细菌学研究中的应用

目前基因芯片技术广泛应用于生物学研究的各个领域,在细菌学研究中基因芯片技术突破了以往单个或数个基因研究的局限,在细菌功能基因组学、药物作用靶点、细菌-宿主细胞间相互作用以及细菌进化等方向发挥着重要作用,初步揭示了细菌生物功能多样性受到多个基因的调控,但相关的研究尚有待深入.     

基因芯片技术及其在疾病基因诊断中的应用

基因芯片技术是伴随着人类基因组计划的实施而发展起来的生命科学领域里的前沿生物技术。它最显著的特点是高通量、高集成、微型化、平行化、多样化和自动化。经过短短十几年的发展,基因芯片技术现已在基因表达分析,基因突变及多态性分析,疾病基因诊断,药物及毒物基因组学等多个领域显示出重大的理论意义和实际应用价值,具有广阔的前景。本文专门介绍了基因芯片技术及其在疾病基因诊断上的应用。     

交联法在分子生物学研究中的应用

本文概要介绍了交联法用于阐明染色质、核小体、病毒粒子、核糖体结构、观察HS70基因-组蛋白等七种核酸-蛋白质复合物中大分子相互作用的实例;列举了它在DNA构象、RNA立体结构及mRNA、tRNA、snRNA、hnRNA、rRNA间相互作用等研宄中的应用。从文中可以看出,交联法的应用范围遍及遗传信息储存的高级结构、转录体系及调控、翻译系统及调控等分子生物学研究的几个主要侧面。     

基因芯片技术在植物基因克隆中的应用研究进展

基因芯片是以预先设计的方式将大量的生物讯息密码(寡核苷酸、cDNA、基因组DNA等)固定在玻片、硅片等固相载体上组成的密集分子阵列.基因芯片技术本质是生物信号的平行分析,它利用核酸分子杂交原理,通过荧光标记技术检测杂交亲和与否,再经过计算机分析处理可迅速获得所需信息.由于其具有高通量、微型化、连续化、自动化、快速和准确等特点,已引起国际国内广泛的关注和重视,在许多领域得到了广泛的应用.本文简述了基因芯片的概念,技术特点及主要分类,着重对其在基因表达水平检测,基因突变和多态性的分析,基因组DNA分析,后基因组学研究以及转基因农作物检测等方面进行阐述,并说明其存在的问题及展望.     

基因芯片及其在环境微生物研究中的应用

基因芯片因其具有高密度、高灵敏度、快速 (实时 )检测、经济、自动化和低背景水平等特点 ,而广泛应用于不同的研究领域。目前 ,应用于环境微生物研究的基因芯片主要有功能基因芯片 (FGAs)、系统发育的寡核苷酸芯片 (POAs)和群落基因组芯片 (CGAs)。综述了基因芯片在环境微生物研究中的应用 ,包括自然环境中微生物的基因表达分析、比较基因组分析和混合微生物群落的分析等。讨论了基因芯片面临的挑战和前景展望     

基因芯片技术在甘蔗研究中的应用进展

基因芯片技术是新生代的生物技术,具有大规模平行处理生命信息的能力.基因芯片具有高通量、并行性、微型化与自动化的特点,因此成为探究功能基因组学最有效的方法之一,已引起全世界广泛的关注和重视,在许多领域得到了广泛的应用.甘蔗是世界上急需研发的重要能源作物,基因芯片对甘蔗研究有重要的意义.本文简介了基因芯片技术的原理和制备过程,并着重阐述了在甘蔗抗旱、抗病、基因表达和miR-NA鉴定方面的应用进展.     

Global analysis of non-coding small RNAs in Arabidopsis in response to jasmonate treatment by deep sequencing technology

In plants, non-coding small RNAs play a vital role in plant development and stress responses. To explore the possible role of non-coding small RNAs in the regulation of the jasmonate (JA) pathway, we compared the non-coding small RNAs between the JA-deficient aos mutant and the JA-treated wild type Arabidopsis via high-throughput sequencing. Thirty new miRNAs and 27 new miRNA candidates were identified through bioinformatics approach. Forty-nine known miRNAs (belonging to 24 families), 15 new miRNAs and new miRNA candidates (belonging to 11 families) and 3 tasiRNA families were induced by JA, whereas 1 new miRNA, 1 tasiRNA family and 22 known miRNAs (belonging to 9 families) were repressed by JA.     

A quantitative trait loci analysis of zinc hyperaccumulation in Arabidopsis halleri

The mechanisms of metal hyperaccumulation are still not understood, so we conducted a quantitative trait locus (QTL) analysis of zinc (Zn) hyperaccumulation in Arabidopsis halleri, in a cross between this and its sister species, A. petraea, in order to determine the number and approximate location of the genomic regions significantly contributing to this adaptation. An F2 cross between the two species was made, and the leaf Zn concentration of 92 individuals was measured at both low (10 microm) and high (100 microm) Zn concentrations. Twenty-five markers were established that were distributed on all of the eight chromosomes. Mapping of the markers established that they were essentially collinear with previous studies. QTLs exceeding a logarithm to the base 10 of the odds (LOD) value of 3 were found on chromosomes 4 (low Zn), 6 (high Zn) and 7 (both high and low Zn). Evidence for a QTL on chromosome 3 (low Zn) was also found. This analysis validates a previously used method of QTL analysis, based on microarray analysis of segregating families. Genes that have altered during the evolution of this character should also be QTL: this analysis calls into question a number of candidate genes from consideration as such primary genes because they do not appear to be associated with QTLs.     

插入诱变在拟南芥基因克隆中的应用

随着各种基因克隆方法的建立 ,克隆的拟南芥基因越来越多 ,其中转座子标签和T DNA插入诱变克隆的拟南芥基因的数目最多 ,插入诱变已成为克隆和鉴定很多重要植物基因的方法。     

Large-scale T-DNA mutagenesis in Arabidopsis for functional genomic analysis

In planta Agrobacterium-mediated transformation combined with a soil-based herbicide selection for transgenic plants was used to recover large numbers of transgenic Arabidopsis plants for functional genomic studies. A tissue-culture-free system for generating transgenic plants was achieved by infiltrating Arabidopsis plants with Agrobacterium tumefaciens harboring a binary T-DNA vector containing the phosphinothricin acetyltransferase gene from Streptomyces hygroscopicus, and by selecting transgenic Arabidopsis growing in soil by foliar application of the herbicide Finale (phosphinothricin). Analysis of herbicide-resistant plants indicated that all were transgenic and that the T-DNA transformation process occurred late during flower development, resulting in a preponderance of independently derived T-DNA insertions. T-DNA insertions were usually integrated in a concatenated, rearranged form, and using linkage analysis, we estimated that T1 plants carried between one and five T-DNA loci. Using pooling strategies, both DNA and seed pools were generated from about 38,000 Arabidopsis plants representing over 115,000 independent T-DNA insertions. We show the utility of these transgenic lines for identifying insertion mutations using gene sequence and PCR-based screening. Electronic Publication     

Mutational analysis of blue-light sensing in Arabidopsis

Blue light regulates many physiological and developmental processes in higher plants through the action of multiple photosensory systems. The analysis of photomorphogenic mutants is leading to a better understanding of how the different photosensory systems mediate the wide range of responses observed in blue light. A review of the current literature on photomorphogenic mutants makes it apparent that redundancies exist in photoreceptor function. For example, many blue-light responses that have been shown to be regulated by a blue-light photosensory system are also under phytochrome control. The study of various light-response mutants suggests that a complex sensory network regulates light-mediated responses. This article attempts to piece together information regarding the sensory systems responsible for blue-light-regulated responses.     

拟南芥抗盐突变体的RAPD分析

以筛选得到可以稳定遗传的抗盐单基因突变体2^#和15^#以及野生型拟南芥为材料进行RAPD分析,150条引物中有3条引物在突变体之间扩增出多态性,不仅证明了DNA水平突变的发生,而且表明它们之间遗传背景相似,是一系列抗盐性不同的近似等位基因系。1条引物在突变体的扩增产物比在野生型的扩增产物多出一个大小约为1200bp的片段,初步认为该片段与抗盐的主效基因有关。     

Population history in Arabidopsis halleri using multilocus analysis

A. halleri is a psuedometallophyte with a patchy distribution in Europe and is often spread by human activity. To determine the population history and whether this history is consistent with potential human effects, we surveyed nucleotide variation using 24 loci from 12 individuals in a large A. halleri population. The means of total and silent nucleotide variation (θ_W) are within the range expected for the species. The population genetic neutrality tests Tajima’s D and Wall’s B had significant composite results rejecting panmixia, and Approximate Bayesian Computation analysis revealed that a subdivision model better explained the variation than the standard neutral model, refugia (or admixture), bottleneck or change of population size models. A categorical regression analysis further supports the subdivision model, and under the subdivision model, the neutrality tests are no longer significant. The best support was for two source populations, a situation consistent with the mixing of two populations possibly mediated by human activity. This scenario might limit the genetic diversity and adaptive potential of the population. The non‐neutral population variation described here should be considered in bioinformatic searches for adaptation.     

拟南芥LEC同源基因的生物信息学分析

用生物信息学方法对拟南芥叶状子叶(LEC)基因的核苷酸序列以及推导氨基酸序列的组成、功能结构域、亲水性等进行了分析。结果表明,拟南芥的LEC蛋白为位于细胞核中的亲水性不稳定蛋白;通过对LEC蛋白的功能结构域的分析发现,LEC1和L1L蛋白中高度保守的区域即为CBF-NFYB-HMF结构域,LEC2和FUSCA3蛋白中高度保守的区域为B3结构域。     

Arabidopsis cell wall proteome defined using multidimensional protein identification technology

With the completion of the sequencing of the Arabidopsis genome and the recent advances in proteomic technology, the identification of proteins from highly complex mixtures is now possible. Rather than using gel electrophoresis and peptide mass fingerprinting, we have used multidimensional protein identification technology (MudPIT) to analyse the "tightly-bound" proteome for purified cell walls from Arabidopsis cell suspension cultures. Using bioinformatics for the prediction of signal peptides for targeting to the secretory pathway and for the absence of ER retention signal, 89 proteins were selected as potential extracellular proteins. Only 33% of these were identified in previous proteomic analyses of Arabidopsis cell walls. A functional classification revealed that a large proportion of the proteins were enzymes, notably carbohydrate active enzymes, peroxidases and proteases. Comparison of all the published proteomic analyses for the Arabidopsis cell wall identified 268 non-redundant genes encoding wall proteins. Sixty of these (22%) were derived from our analysis of tightly-bound wall proteins.     

Cell cycle controls: genome-wide analysis in Arabidopsis

The first decade of molecular analysis of plant cell cycle control genes revealed how well the important regulators are conserved among eukaryotes. The recent completion of the Arabidopsis genome sequence, and the use of increasingly sophisticated biochemical assays and genetic approaches, heralds a period of more detailed functional analysis of cell cycle regulators aimed at resolving their role in plant growth and development.