转座因子标签法

转座因子(Transposable element)是Mc-Clintock在玉米的染色体上首先发现的,以后在大肠杆菌、酵母、果蝇、线虫、蚕及金鱼草与矮牵牛等植物中也陆续发现了转座因子的存在。转座因子的发现是遗传学发展史上的重要里程碑,是本世纪遗传学领域内重大的发现之一,在理论和实际应用上都有重要的意义。近年来,由于分子遗传学研究的进步,对转座因子的结构、转座的机理等方面的研究取得了很大的成绩,转座因子的应用研究开始受到人们的重视,特别在应用转座因子作为标签来分离高等植物基因的研究中取得了令人瞩目的成绩。本文就转座因子标签法分离高等植物基因的研究进展作一大致的介绍。     

杆状病毒转座因子研究进展

转座因子(Transposable elements)是一类可移动转座的遗传因子的统称,包括原核生物中的插入(IS)、转座子(Tn)、质粒;真核生物中的Ty,P因子,2μDNA,Copia因子,以及噬菌体Mu和反转录病毒等。因此,转座因子又称可移动的遗传因子(Mobile genetic element)。转座因子最早由美国科学家Barbara McClintock于1956年在玉米染色体中发现,并于1984年被授予诺贝尔医学或生理学奖。转座因子的发现无论在理论上还是实践上都具有很重要的意义,被认为是遗传学发展史上的重要里程碑之一。杆状病毒是一类以节肢动物(主要发现于昆虫纲鳞翅目)为宿主的病毒的统称。杆状病毒亦存在转座因子。对杆状病毒转座因子的研究起源于对感     

植物可移动遗传因子研究进展

本文介绍了植物中转座因子的遗传特性和转座因子的分子结构特点以及转座的调节控制。植物中的转座因子一般为1. 4-17kb的DNA分子,本身具有末端重复顺序,在插人位点造成DNA重复。转座的功能由转座因子本身决定,玉米的Ac因子和Spm因子都已证明带有决定转座的基因,这些基因的表达调节以及基因产物如何发挥作用仍在研究之中。文中还讨论了转座因子的作用和一些尚未解决的问题。     

植物可移动遗传因子研究进展

本文介绍了植物中转座因子的遗传特性和转座因子的分子结构特点以及转座的调节控制。植物中的转座因子一般为1. 4-17kb的DNA分子，本身具有末端重复顺序，在插人位点造成DNA重复。转座的功能由转座因子本身决定，玉米的Ac因子和Spm因子都已证明带有决定转座的基因，这些基因的表达调节以及基因产物如何发挥作用仍在研究之中。文中还讨论了转座因子的作用和一些尚未解决的问题。     

转座元件、表观遗传调控与细胞命运决定

转座元件是哺乳动物基因组内含量最多的元素。尽管转座元件的存在对基因组稳定性具有潜在的危险,但它们同时还是潜在的基因调控序列、蛋白质编码序列和染色质结构序列,并参与物种进化过程。因此,基因组中转座元件的有害性和有益性保持着谨慎的平衡,并且这种平衡主要由表观遗传修饰来调控。本文详细介绍了异染色质类型表观遗传修饰如H3K9me3和DNA甲基化在转座元件沉默中的功能;转座元件作为增强子元件富集激活型表观遗传修饰如H3K4me1和H3K27ac,以及作为转录因子结合靶点、染色质构象锚点等方式参与基因表达调控的模式;从体内胚胎发育到体外细胞命运转变,阐述了转座元件在细胞命运决定中的潜在功能及作用方式;最后,对转座元件领域研究存在的挑战及潜在解决方法提出了见解。总之,本文对转座元件与表观遗传、基因表达调控以及细胞命运决定等方面的研究及存在的问题进行了较全面的综述,旨在为相关领域的研究人员提供参考。     

科学出版社生命科学编辑部新书推介

本书是现代遗传学丛书之一。第三版中除删去第二版中的经典遗传学概论一章,把遗传的物质基础一章并人绪论,把形态建成和分化发育一章的内容并人有关章节,并在非孟德尔式遗传学一章增加转座因子和遗传工程概要两节的内容以外其余部分基本上保持原貌,作为本书的上篇。     

植物转座因子

转座因子 (ｔｒａｎｓｐｏｓａｂｌｅｅｌｅｍｅｎｔ ,ＴＥｓ)是指在生物细胞中能从同一条染色体的一个位点转移到另一个位点或者从一条染色体转移到另一条染色体上的ＤＮＡ序列。 1 947年美国冷泉港实验室的“玉米夫人”ＭｃＣｌｉｎｔｏｃｋ首先在玉米中发现并描述了转座因子。转座因子的发现 ,打破了传统遗传学上关于基因在染色体上固定排列及同源染色体交换的观念 ,揭示了基因的流动性 ,具有重要的意义。1 .转座因子的结构特点和分类到目前报道为止 ,至少在 32种植物上有转座因子存在 ,其中研究最多的是玉米、金鱼草、拟南芥等[1] 。其…     

可动因子与生物进化

可动因子与生物进化李宏（渝州大学生物系,重庆,６３００３３）自从１９８３年,美国遗传学家ＭｃＣｌｉｎｔｏｃｋ在玉米籽粒的色斑遗传研究中发现了转座因子（ｔｒａｎｓｐｏｓｏｎ）以来,人们逐渐发现除了这种转座因子之外,还存在着另外一些能发生转座的因子,比如...     

《中国大百科全书生物学》卷遗传学条目—转座因子

转座因子（transposable element） 细 胞中能改变自身位置的一段脱氧核塘核酸(DNA）序 列。转座因子改变位置（例如从染色体上的一个位置 转移到另一个位置,或者从质粒转移到染色体上）的行 为称为转座。 第一个转座因子是四十年代美国遗传学家B.麦 克林托克在玉米中发现的解离因子（见位置效应）。现 在证明果蝇、啤酒酵母(Saccharomyces cerevisiae）与大 肠杆菌(Escherichia codi等的染色体以及多种细菌质 粒上也都有不同类别的转座因子存在,不但某些噬菌 体DNA本身就是转座因子,而且有些致癌的RNA病 毒的前病毒也具有细菌转座子的结构。     

果蝇转座因子对基因组进化的影响

真核生物基因组织有很多可移动ＤＮＡ片段为称转座因子,果蝇是大量系统研究的最好实验材料之一,其基因组的１０％－１２％是由转座因子组成,在宿主中,ＴＥs也许改变基因表达模型,也许改变ＯＲＦs编码序列,也许对细胞功能产生影响,这此因子遗传的可动性也可能使它们适于建造载体产生转基因生物。因此,对ＴＥs进化的动态研究以及对宿主基因组进化影响的探索将有助于TEs作为载体的细胞工程研究。     

Development and use of an efficient system for random mariner transposon mutagenesis to identify novel genetic determinants of biofilm formation in the core Enterococcus faecalis genome

Enterococcus faecalis is a gram-positive commensal bacterium of the gastrointestinal tract and an important opportunistic pathogen. Despite the increasing clinical significance of the enterococci, most of the genetic analysis of these organisms has focused on mobile genetic elements, and existing tools for manipulation and analysis of the core E. faecalis chromosome are limited. We are interested in a comprehensive analysis of the genetic determinants for biofilm formation encoded within the core E. faecalis genome. To identify such determinants, we developed a substantially improved system for transposon mutagenesis in E. faecalis based on a mini-mariner transposable element. Mutagenesis of wild-type E. faecalis with this element yielded predominantly mutants carrying a single copy of the transposable element, and insertions were distributed around the entire chromosome in an apparently random fashion. We constructed a library of E. faecalis transposon insertion mutants and screened this library to identify mutants exhibiting a defect in biofilm formation. Biofilm-defective mutants were found to carry transposon insertions both in genes that were previously known to play a role in biofilm formation and in new genes lacking any known function; for several genes identified in the screen, complementation analysis confirmed a direct role in biofilm formation. These results provide significant new information about the genetics of enterococcal biofilm formation and demonstrate the general utility of our transposon system for functional genomic analysis of E. faecalis.     

Regulation of Drosophila P element transposition.

Drosophila P transposable elements are the best-studied family of eukaryotic non-retroviral transposons. P element transposition is regulated in several different ways and has thus provided a unique system with which to study the control of DNA rearrangements and gene expression in metazoans. Recent genetic and biochemical experiments have begun to shed light on the mechanism of P element transposition and the mechanisms controlling the temporal and spatial patterns of transposition.     

A P Element Containing Suppressor of Hairy-Wing Binding Regions Has Novel Properties for Mutagenesis in Drosophila Melanogaster

P elements are widely used as insertional mutagens to tag genes, facilitating molecular cloning and analyses. We modified a P element so that it carried two copies of the suppressor of Hairy-wing [su(Hw)] binding regions isolated from the gypsy transposable element. This transposon was mobilized, and the genetic consequences of its insertion were analyzed. Gene expression can be altered by the su(Hw) protein as a result of blocking the interaction between enhancer/silencer elements and their promoter. These effects can occur over long distances and are general. Therefore, a composite transposon (SUPor-P for suppressor-P element) combines the mutagenic efficacy of the gypsy element with the controllable transposition of P elements. We show that, compared to standard P elements, this composite transposon causes an expanded repertoire of mutations and produces alleles that are suppressed by su(Hw) mutations. The large number of heterochromatic insertions obtained is unusual compared to other insertional mutagenesis procedures, indicating that the SUPor-P transposon may be useful for studying the structural and functional properties of heterochromatin.     

What is the impact of transposable elements on host genome variability?

The spread of a transposable element family through a wild population may be of astonishing rapidity. At least three families of transposable genetic elements have recently invaded Drosophila melanogaster worldwide, including the P element. The mechanism has been a process of effectively replicative transposition, and, for the P element, has occurred notwithstanding the sterility induced by unrestricted movement. This element's invasion into D. melanogaster has been accompanied by the development of heterogeneity between P sequences, most of which now have internal deletions. Increasing evidence suggests that some deleted elements can repress P transposition, thereby protecting the host from the harmful effects of complete elements. Such repressing elements may rise to high frequencies in populations as a result of selection at the level of the host. We here investigate selective sweeps invoked by the spread of P sequences in D. melanogaster populations. Numerous high-frequency sites have been identified on the X chromosome, which differ in frequency between populations, and which are associated with repression of P-element transposition. Unexpectedly, sequences adjacent to high-frequency P-element sites do not show reduced levels of genetic diversity, and DNA variability is in linkage equilibrium with the presence or absence of a P element at the adjacent selected site. This might be explained by multiple insertions or through a selection for recombination analogous to that seen in 'hitchhiking'.     

Genetic transformation of Populus genotypes with different chimaeric gene constructs: transformation efficiency and molecular analysis

Aspen (Populus tremula) and hybrid aspen (P. tremula × P. tremuloides) were transformed with different gene constructs using two types of promoter. The aim was to determine the influence of the reporter gene rolC, controlled by promoters of viral or plant origin, on genetic and morphologic expression of different transgenic aspen clones. An improved transformation method using leaf discs was developed, by which putative transgenic plantlets were regenerated at high efficiencies (up to 34%) on kanamycin-containing medium. Transgenic aspen carrying the rolC gene from Agrobacterium rhizogenes under control of the cauliflower-35S-promoter are reduced in size with smaller leaves, whereas aspen transgenic for the same rolC gene, but under control of the light inducible rbcS promoter from potato, are only slightly reduced in size compared to untransformed controls. However, all clones carrying 35S-rolC and rbcS-rolC genes revealed light-green colouration of leaves when compared to untransformed aspen. Owing to this special feature, constructs were used in which expression of the rolC gene was inhibited by insertion of a transposable element, Ac, from maize. Transgenic aspen transformed with the 35S-Ac-rolC and rbcS-Ac-rolC genes were morphologically similar to untransformed aspen, but out of 54 independently regenerated 35S-Ac-rolC transgenic aspen clones, 30 clones showed light-green/dark green variegated leaves. In contrast, out of 19 independently transformed rbcS-Ac-rolC aspen clones, only two clones revealed light-green/dark green variegated leaves. The role of bacterial strains in transformation, and molecular genetics of transgenic aspen plants (including the function of the transposable element, Ac, in the aspen genome) are discussed     

Isolation of Mutations in the Drosophila Homologues of the Human Neurofibromatosis 2 and Yeast Cdc42 Genes Using a Simple and Efficient Reverse-Genetic Method

Reverse genetic analysis in Drosophila has been greatly aided by a growing collection of lethal P transposable element insertions that provide molecular tags for the identification of essential genetic loci. However, because the screens performed to date primarily have generated autosomal P-element insertions, this collection has not been as useful for performing reverse genetic analysis of X-linked genes. We have designed a reverse genetic screen that takes advantage of the hemizygosity of the X chromosome in males together with a cosmid-based transgene that serves as an autosomally linked duplication of a small region of the X chromosome. The efficacy and efficiency of this method is demonstrated by the isolation of mutations in Drosophila homologues of two well-studied genes, the human Neurofibromatosis 2 tumor suppressor and the yeast CDC42 gene. The method we describe should be of general utility for the isolation of mutations in other X-linked genes, and should also provide an efficient method for the isolation of new alleles of existing X-linked or autosomal mutations in Drosophila.     

Transposable element-medicated evolution of sex: A population genetic model

For a population made up of individuals capable of sexual as well as asexual modes of reproduction, conditions for the spread of a transposable element are explored using a one-locus, two-haplotype model. The analysis is then extended to include the possibility that the transposable element can modulate the probability of sexual reproduction, thus casting Hickey’s (1982,Genetics 101: 519–531) suggestion in a population genetics framework. The model explicitly includes the cost of sexual reproduction, fitness disadvantage to the transposable element, probability of transposition, and the predisposition for sexual reproduction in the presence and absence of the transposable element. The model predicts several kinds of outcome, including initial frequency dependence and stable polymorphism. More importantly, it is seen that for a wide range of parameter values, the transposable element can go to fixation. Therefore it is able to convert the population from a predominantly asexual to a predominantly sexual mode of reproduction. Viewed in conjunction with recent results implicating short stretches of apparently non-coding DNA in sex determination (McCoubreyet al. 1988,Science 242: 1146–1151), the model hints at the important role this mechanism could have played in the evolution of sexuality.     

Asymmetric exchange is associated with P element induced male recombination in Drosophila melanogaster

Spontaneous meiotic recombination events do not normally occur in the male germ line of Drosophila melanogaster. However, such events are induced in males when a P transposable element or a source of P element encoded transposase protein is present in its genome. This report concerns a molecular analysis of the meiotic exchanges that were induced in the male Drosophila by P elements within a genetically marked region of the third chromosome. The marked region also harbors a single P-element called P(lArB). Fifty-six percent of the P(lArB) region crossovers indicated some alterations in the P element 5' fragment. Such alterations appear to be related to asymmetric or unequal genetic exchanges. Finally, P(lArB) excision was found to be independent of P(lArB) region crossover events.     

PCR detection of excision suggests mobility of the medaka fish Tol1 transposable element in the frog Xenopus laevis

Tol1 is a DNA-based transposable element identified in the medaka fish Oryzias latipes and a member of the hAT (hobo/Activator/Tam3) transposable element family. Its mobility has already been demonstrated in the human and mouse, in addition to its original host species. This element is thus expected to be useful in a wide range of vertebrates as a genomic manipulation tool. Herein, we show that the Tol1 element can undergo excision in the African clawed frog Xenopus laevis, a major model organism for vertebrate genetics and developmental biology. An indicator plasmid carrying a Tol1 element was injected into 2- or 4-cell-stage embryos together with either a helper plasmid coding for the full-length Tol1 transposase or a modified helper plasmid yielding a truncated protein, and recovered from tailbud-stage embryos. Deletion of the Tol1 region of the indicator plasmid was observed in the experiment with the full-length transposase, and not in the other case. The deletion was associated with various footprint sequences at breakpoints, as frequently observed with many DNA-based transposable elements. These results indicate that the Tol1 element was excised from the indicator plasmid by catalysis of the transposase, and suggest that the Tol1 element is mobile in this frog species.