共查询到19条相似文献,搜索用时 203 毫秒
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1 转座子及转座子标签法克隆基因基因标签法克隆植物组织中的基因是较为常用的一种方法 ,T-DNA和转座子均可作为基因标签。转座子最早由美国的细胞遗传学家 Mc-clintock在玉米中发现 ,它是指基因组中一段特定 DNA片段 ,能在转位酶的作用下从基因组的一个位点转移到另一个位点。转座子不仅能在本基因组中转座 ,也能转入其它植物的基因组中。转座的结果是使被转入的基因失活 ,从而有效地诱导产生表型突变株。然后构建一个对应于突变株的基因库 ,用作标签的转座子作为探针从基因库中筛选相对应的克隆 ,分离得到相对应于变异的基因 ,这就是… 相似文献
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植物基因工程中的转座子标签 总被引:9,自引:1,他引:8
195 1年BarbaraMclintock首先在玉米中发现了控制元件 ,后来命名为转座元件或转座子 (transposon)。转座子是基因组中一段可移动的DNA序列 ,可以通过切割、重新整合等一系列过程从基因组的一个位置“跳跃”到另一个位置。这一元件不仅可用于分析生物遗传进化上分子作用引起的一些现象 ,还为基因工程和分子生物学研究提供了强有力的工具 ,可以在不了解基因产物的生化性质和表达模式的情况下 ,分离克隆植物基因 ,即转座子标签 (transposontagging) ,又称为转座子示踪法。其原理是利用转座子的… 相似文献
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植物转座子及其在功能基因组学中的应用 总被引:14,自引:3,他引:11
转座子作为插入突变原或分子标签被广泛应用于基因的分离和克隆,且因其
独特的性质已成为发现新基因和基因功能分析的有效工具。本文综述了植物转座子及其作为基因分离和克隆工具的研究进展,并讨论其在植物基因功能研究方面的应用。
Abstract:Transposons have been widely used for gene isolation and gene cloning as insertional mutagens or molecular tagging.Furthermore,due to special characteristics of transposons,transposons techniques will be a powerful tool for new gene discovery and gene functional analysis.This paper reviewed the developments of plant transposons in gene isolating and cloning,as well as its use in studying gene function in plant. 相似文献
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本文综述了植物转座子在基因工程应用上的最新研究进展。转座子已成为引入注目的植物基因分离手段。转座子还可以作为一种新的载体系统介导外源基因转移并消除转基因植物中的选择标记基因。 相似文献
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转座子是DNA插入因子的一种,是指能在基因组间或组内跳跃的DNA片段。转座子作为插入突变剂或分子标签已被广泛地应用于基因的分离和克隆,且因其独特的性质已成为发现新基因和基因功能分析的有效工具。这使得转座子无论是在单基因水平还是全基因组水平,都成为细菌、酵母和其他微生物研究的有力工具。简单而有效的体外转座反应可以对一些以往难以进行分析的顽固微生物进行转座诱变分析。而建立在转座子基础上的信号标签诱变技术和遗传足迹法的应用则发现了一些新的病原微生物毒力因子,从而可以更好地对这些病原微生物的致病机理进行阐述。这些再次说明转座子是微生物功能基因组研究中的有力工具。本文综述了转座子及其衍生载体介导的一些技术,并讨论其在微生物功能基因组研究中的应用。 相似文献
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植物抗病基因克隆研究进展 总被引:1,自引:0,他引:1
随着分子生物学及其相关技术的飞速发展,人们对植物与病原微生物相互作用的分子机制了解得越来越透彻。本文对植物过敏性反应和系统获得抗性作了简要概述,并着重讨论了植物抗病基因克隆的进展,涉及到转座子标签技术、定位克隆技术、染色体步行、染色体登陆等方法和策略,归纳了克隆到的植物抗病基因及其产物结构,概述了这些基因产物所共有的特点,并简要介绍了植物抗病基因工程的进展。 相似文献
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转座子标签及其在酿酒酵母基因功能研究中的应用 总被引:2,自引:0,他引:2
转座子标签(transposontagging)技术是研究功能基因的有效的工具之一。介绍了几种在酿酒酵母(Saccharomycescerevisiae)基因功能研究中应用的转座子标签:mTn3标签、miniMu噬菌体标签和Ty转座子标签,阐述了转座子标签的构建原理、应用策略和转座子标签插入位点的鉴定方法。 相似文献
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Joel M. Chandlee 《Physiologia plantarum》1990,79(1):105-115
Transposable elements are segments of DNA which have the unique capability of being able to excise from one site in the genome and reintegrate into new, different sites elsewhere in the genome. When transposition takes place and integration occurs within a gene locus, mutations are frequently generated producing variegated or recessive phenotypes. This ability of transposable elements to act as mutagenic agents through their association with particular gene sequences has lead to the development of the procedure of transposon tagging or gene tagging in higher plants. Through this technique, transposable elements can be used to clone and isolate genes of interest for which little or nothing is known about the final product (i.e., polypeptide). This offers tremendous potential for the isolation of a variety of agronomically important genes, which are virtually impossible to recover by other currently available gene cloning methodologies. To date, the technique has been used successfully to isolate genes from corn and snapdragon. Using gene transfer technologies, the potential now exists to extend this approach to clone genes from other plant species. Advantages and limitations of transposon tagging for isolating plant genes will be discussed. 相似文献
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Colwyn M. Thomas Pieter Vos Marc Zabeau David A. Jones Karen A. Norcott Brian P. Chadwick Jonathan D.G. Jones 《The Plant journal : for cell and molecular biology》1995,8(5):785-794
Using the technique of amplified restriction fragment polymorphism (AFLP) analysis, and bulked segregant pools from F2 progeny of the cross Lycopersicon esculentum (Cf9)× L. pennellii , approximately 42 000 AFLP loci for tight linkage to the tomato Cf-9 gene for resistance to Cladosporium fulvum have been screened. Analysis of F2 recombinants identified three markers which co-segregated with Cf-9 . The Cf-9 gene has recently been isolated by transposon tagging using the maize transposon Dissociation ( Ds ). Analysis of plasmid clones containing Cf-9 shows that two of these markers are located on opposite sides of the gene separated by 15.5 kbp of intervening DNA. AFLP analysis provides a rapid and efficient technique for detecting large numbers of DNA markers and should expedite plant gene isolation by positional cloning and the construction of high-density molecular linkage maps of plant genomes. 相似文献
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L.J.G. van Enckevort J. E. M. Bergervoet W. J. Stiekema A. Pereira E. Jacobsen 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》2000,101(4):503-510
Potato is an autotetraploid crop plant that is not very amenable to the deployment of transposon tagging for gene cloning
and gene identification. After diploidisation it is possible to get potato genotypes that grow well, but they are self-incompatible.
This prevents the production of selfed progeny that are normally used in gene tagging approaches to select for parental lines
with the target gene to be tagged in a homozygous stage. We describe here an alternative selection method for directed transposon
tagging for a gene of interest in a heterozygous background. Diploid potato plants with a Ds transposon linked to the desired gene of interest (the Phytophthora
infestans R1 resistance locus) in a heterozygous stage were used for the development of this directed transposon tagging strategy. After
crossing to a diploid Ac transposon-containing genotype, 22 ’interesting’ seedlings (R1Ds/r–; Ac/–) were selected that showed active Ds transposition as displayed by DNA blot hybridisation, empty donor site PCR and sequencing. Protoplast isolation and the use
of the hygromycin gene as a cell-specific selection marker of Ds excision enabled the direct selection of Ds excision sectors in these highly chimaeric seedlings. This somatic selection of Ds transpositions and the regeneration through protoplasts resulted in the development of a large population of almost 2000
hygromycin-resistant plants. Southern blot analysis confirmed the insertion of Ds at independent positions in the genome. Every selected plant displayed independent Ds excisions and re-insertions due to the expression of the Ac transposase throughout development. This population, which is developed from seedlings with the desired R1 gene in a heterozygous stage, is directly useful for searching for transposon-tagged R1 mutants. In general, this approach for selecting for somatic transpositions is particularly suitable for the molecular isolation
of genes in a heterozygous crop like potato.
Received: 29 November 1999 / Accepted: 30 December 1999 相似文献
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Summary This article reviews techniques for gene identification and cloning in allohexaploid bread wheat (Triticum aestivum L.). Gene identification and cloning in wheat are complicated by the large size and high redundancy of the genome. Both classical
mutagenesis and transposon tagging are important tools for the study of grain dormancy and plant hormone signaling in wheat.
While classical mutagenesis can be used to identify wheat mutants with altered hormone sensitivity, it can be difficult to
clone the corresponding genes. We review the techniques available for gene identification in wheat, and propose that transposon-based
activation tagging will be an important tool for wheat genetics. 相似文献
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