分子生物学教材中的真核生物基因组重复序列

现行的高校分子生物学教材中主要以重复频率为依据对重复序列进行分类,对于小卫星DNA及微卫星DNA是属于高度或是中度重复序列存在不同见解。提出依据重复频率及空间结构分布两个方面对重复序列进行分类,并建议按照重复频率将小卫星DNA及微卫星DNA归属于中度重复序列。     

一个AA基因组特异的串联重复序列的克隆及其在中国普通野生稻和栽？…

从籼稻“窄叶青”中克隆到了１个重复序列（ｐＯｓ１３９）。经分子杂交证明,ｐＯｓ１３９为一稻属内ＡＡ基因组特异的串联重复序列。序列分析表明,ｐＯｓ１３９以３５５ｂｐ为一重复单位。以ｐＯｓ１３９为探针对２９份中国普通野生稻和４３份中国栽培稻的基因组ＤＮＡ进行的分子杂交表现,籼、粳亚种之间具有明显的差异,籼稻杂交带数明显多于粳稻,普通野生稻与籼稻相似,具有较多的杂交带数。拷贝数测定结果表明,ｐＯｓ１３９     

串联重复序列在琴叶拟南芥基因组中的特征探究

串联重复序列广泛存在于真核生物的基因组中,它通过影响染色质的空间结构及基因表达从而影响生物的遗传与进化.本研究以琴叶拟南芥(Arabidopsis lyrata)基因组为材料,分析了1～50 bp重复单元的串联重复序列特征.研究发现串联重复序列在基因的5'UTR和启动子区域密度最高(8757 bp/Mb,8430 bp/Mb),而编码区CDS的密度最低(2406 bp/Mb).基因组中重复模体最高的为单核苷酸重复的T/A碱基,5'UTR中包含大量的二核苷酸重复模体,而在CDS中主要是三核酸重复模体.串联重复序列特征在琴叶拟南芥基因组不同区域的差别,显示其与基因表达和调控功能相适应.本研究深入探讨了串联重复序列在植物基因组中的特征及作用,为重复序列调控基因表达及植物基因组进化提供借鉴.     

卫星,小卫星和微卫星DNA—真核生物基因组的串状重复序列

卫星、小卫星和微卫星ＤＮＡ——真核生物基因组的串状重复序列姜运良（山东农业大学动物科技学院,山东泰安２７１０１８）关键词卫星小卫星微卫星串状重复序列真核生物基因组中编码蛋白质（酶）的结构基因只占很少的一部分（１０％～２０％）,其余大部分是重复序列。根...     

大卫鼠耳蝠线粒体控制区串联重复序列变异

中国特有种大卫鼠耳蝠线粒体D-loop区串联重复序列以81bp为重复单元,重复3～7次。54%的个体具有4个串联重复序列,与5～7个串联重复序列的现有模式具有较大的差异。重复单元的遗传差异受所在位次影响,具有明显的家族性或区域性特点。串联重复序列和相对保守的第一重复单元构建的ML树均形成了3个明显的分支,分别定名为东南区、西南区和南方区。线粒体重复序列区域间的差异暗示其可能经历了多次进化,并以东南区变异最为显著。     

中国明对虾基因组小卫星重复序列分析

通过对中国明对虾基因组随机DNA片断的测序 ,我们获得了总长度约 6 4 10 0 0个碱基的基因组DNA序列 ,从中共找到 172 0个重复序列。其中 ,小卫星序列的数目为 398个 ,占重复序列总数目的 2 3 14 %。这些小卫星序列的重复单位长度为 7- 16 5个碱基 ,集中分布于 7- 2 1个碱基范围内 ,其中以重复单位长度为 12个碱基的重复序列数目最多 ,为 5 8个 ,占小卫星重复序列总数目的 14 5 7%。不同拷贝数目所对应的重复序列的数目情况为 :拷贝数目为 2的重复单位所组成的重复序列数目最多 ,为 137个 ;其次是拷贝数目为 3的重复序列 ,为12 2个 ,且随着拷贝数目的增加 ,由其所组成的重复序列的数目呈递减的趋势。其中一部分序列见GeneBank数据库 ,登录号为AY6 990 72 -AY6 990 76。 398个重复序列分别由 398种重复单位所组成 ,因而小卫星重复序列的类型很多 ,我们初步分成三类 :两种碱基组成类别、三种碱基组成类别和四种碱基组成类别 ,并进一步根据各个重复序列中所含有的碱基种类的数量从大到小排列这些碱基而分成若干小类。从这些分类中可以看出 ,中国明对虾基因组中的小卫星整体上是富含A T的重复序列 ,并具有一定的“等级制度” ,揭示了其与微卫星重复序列之间的关系 ,即一部分小卫星重复序列可能起源于微卫星     

中华鲟(Acipenser sinensis)及相关种类的mtDNA控制区串联重复序列及其进化意义

采用 PCR技术和 DNA测序技术 ,发现了我国一级珍稀保护动物中华鲟 ( Acipensersinensis)线粒体 DNA( mt DNA)的控制区 ( D- loop)存在数目不等的串联重复序列 ,该重复序列造成了中华鲟广泛的异质性现象 .从分子水平进行了不同类型重复序列变化规律的研究 ,同时还初探了重复序列在我国其它几种鲟鱼类的存在情况 ,发现在白鲟 ( Psephurus gladius)、达氏鲟 ( A.dabryanus)和史氏鲟 ( A.schrenckii)均存在类似的重复序列结构 .序列比较分析表明 ,不同鲟鱼类重复序列在鲟鱼类进化过程中扮演着一定的角色 ,很有可能碱基差异大小与它们的亲缘关系的远近呈正相关 .     

一个AA基因组特异的串联重复序列的克隆及其在中国普通野生稻和栽培稻中的分化特征

从籼稻（ＯｒｙｚａｓａｔｉｖａＬ．ｓｐｐ．ｉｎｄｉｃａ）“窄叶青”中克隆到了１个重复序列（ｐＯｓ１３９）。经分子杂交证明,ｐＯｓ１３９为一稻属内ＡＡ基因组特异的串联重复序列。序列分析表明,ｐＯｓ１３９以３５５ｂｐ为一重复单位。以ｐＯｓ１３９为探针对２９份中国普通野生稻和４３份中国栽培稻的基因组ＤＮＡ进行的分子杂交表明,籼、粳亚种之间具有明显的差异,籼稻杂交带数明显多于粳稻,普通野生稻与籼稻相似,具有较多的杂交带数。拷贝数测定结果表明,ｐＯｓ１３９在普通野生稻和籼稻中丰度均较高,在粳稻中丰度较低。结合ｐＯｓ１３９的Ｓｏｕｔｈｅｒｎ杂交结果和以前的ＲＡＰＤ结果,认为籼稻和粳稻共同起源于普通野生稻。     

Retrotransposons and Tandem Repeat Sequences in the Nuclear Genomes of Cryptomonad Algae

The cryptomonads are an enigmatic group of unicellular eukaryotic algae that possess two nuclear genomes, having acquired photosynthesis by the uptake and retention of a eukaryotic algal endosymbiont. The endosymbiont nuclear genome, or nucleomorph, of the cryptomonad Guillardia theta has been completely sequenced: at only 551 kilobases (kb) and with a gene density of ∼1 gene/kb, it is a model of compaction. In contrast, very little is known about the structure and composition of the cryptomonad host nuclear genome. Here we present the results of two small-scale sequencing surveys of fosmid clone libraries from two distantly related cryptomonads, Rhodomonas salina CCMP1319 and Cryptomonas paramecium CCAP977/2A, corresponding to ∼150 and ∼235 kb of sequence, respectively. Very few of the random end sequences determined in this study show similarity to known genes in other eukaryotes, underscoring the considerable evolutionary distance between the cryptomonads and other eukaryotes whose nuclear genomes have been completely sequenced. Using a combination of fosmid clone end-sequencing, Southern hybridizations, and PCR, we demonstrate that Ty3-gypsy long-terminal repeat (LTR) retrotransposons and tandem repeat sequences are a prominent feature of the nuclear genomes of both organisms. The complete sequence of a 30.9-kb genomic fragment from R. salina was found to contain a full-length Ty3-gypsy element with near-identical LTRs and a chromodomain, a protein module suggested to mediate the site-specific integration of the retrotransposon. The discovery of chromodomain-containing retroelements in cryptomonads further expands the known distribution of the so-called chromoviruses across the tree of eukaryotes. [Reviewing Editor: Dr. Debashish Bhattacharya]     

Repetitive Sequences in Plant Nuclear DNA:Types,Distribution, Evolution and Function

Repetitive DNA sequences are a major component of eukaryotic genomes and may account for up to 90% of the genome size. They can be divided into minisatellite, microsatellite and satellite sequences. Satellite DNA sequences are considered to be a fast-evolving component of eukaryotic genomes, comprising tandemly-arrayed, highly-repetitive and highly-conserved monomer sequences. The monomer unit of satellite DNA is 150–400 base pairs(bp) in length.Repetitive sequences may be species- or genus-specific, and may be centromeric or subtelomeric in nature. They exhibit cohesive and concerted evolution caused by molecular drive, leading to high sequence homogeneity. Repetitive sequences accumulate variations in sequence and copy number during evolution, hence they are important tools for taxonomic and phylogenetic studies, and are known as ‘‘tuning knobs' ' in the evolution. Therefore, knowledge of repetitive sequences assists our understanding of the organization, evolution and behavior of eukaryotic genomes. Repetitive sequences have cytoplasmic, cellular and developmental effects and play a role in chromosomal recombination. In the post-genomics era, with the introduction of next-generation sequencing technology, it is possible to evaluate complex genomes for analyzing repetitive sequences and deciphering the yet unknown functional potential of repetitive sequences.     

Divergent microsatellite evolution in the human and chimpanzee lineages

Comparison of the complete human genome sequence to one of its closest relatives, the chimpanzee genome, provides a unique opportunity for exploring recent evolutionary events affecting the microsatellites in these species. A simple assumption on microsatellite distribution is that the total length of perfect repeats is constant compared to that of imperfect ones regardless of the repeat sequence. In this paper, we show that this is valid for most of the chimpanzee genome but not for a number of human chromosomes. Our results suggest accelerated evolution of microsatellites in the human genome relative to the chimpanzee lineage.     

Dynamics of Tandem Repetitive Afa-Family Sequences in Triticeae, Wheat-Related Species

The Afa-family sequences in wheat-related species, Triticeae, are tandem repetitive sequences of 340 bp. All the analyzed Triticeae species carried the sequences in their genomes, though the copy numbers varied about 100-fold among the species. The nucleotide fragments amplified by PCR were cloned and sequenced, and their behavior in the evolution of Triticeae was analyzed by the neighbor-joining (NJ) method. The sequences in genomes with many copies of this family clustered at independent branches of the phylogenic tree, whereas the sequences in genomes with a few copies did not. This may suggest that Afa-family sequences had amplified several times in the evolution of Triticeae, each using a limited number of different master copies. In addition, the sequences of the A and B genomes of hexaploid common wheat indicated that the Afa-family sequences had not evolved in a concerted manner between the genomes. Furthermore, the sequences of each chromosome of the D genome of this species indicated that the sequences had amplified on all over the D-genome chromosomes in a short period. Received: 1 September 1997 / Accepted: 19 January 1998     

Heterogeneous Nature and Distribution of Interruptions in Dinucleotides May Indicate the Existence of Biased Substitutions Underlying Microsatellite Evolution

Some aspects of microsatellite evolution, such as the role of base substitutions, are far from being fully understood. To examine the significance of base substitutions underlying the evolution of microsatellites we explored the nature and the distribution of interruptions in dinucleotide repeats from the human genome. The frequencies that we inferred in the repetitive sequences were statistically different from the frequencies observed in other noncoding sequences. Additionally, we detected that the interruptions tended to be towards the ends of the microsatellites and 5'-3' asymmetry. In all the estimates nucleotides forming the same repetitive motif seem to be affected by different base substitution rates in AC and AG. This tendency itself could generate patterning and similarity in flanking sequences and reconcile these phenomena with the high mutation rate found in flanking sequences without invoking convergent evolution. Nevertheless, our data suggest that there is a regional bias in the substitution pattern of microsatellites. The accumulation of random substitutions alone cannot explain the heterogeneity and the asymmetry of interruptions found in this study or the relative frequency of different compound microsatellites in the human genome. Therefore, we cannot rule out the possibility of a mutational bias leading to convergent or parallel evolution in flanking sequences.     

Characterization of Novel Minisatellite Repeat Loci in Atlantic Salmon (Salmo salar) and Their Phylogenetic Distribution

We present here the sequence and characterization of various minisatellite-like tandem repeat loci isolated from the genome of Atlantic salmon (Salmo salar). Their diversity of sequence and lack of core motifs common to minisatellites of other species suggest the presence of numerous and previously unidentified simple sequence repeat families in this salmonid. Evidence for their ubiquity was provided by screening of a salmon genomic library. Southern blot analysis of the phylogenetic distribution of a subset of the minisatellites found one sequence to be pervasive among vertebrates, others present only in Salmoninae or Salmonidae species, and one amplified only in Atlantic salmon. There is evidence for the positioning of microsatellite and minisatellite arrays in close proximity at many loci. Furthermore, one tandem repeat appears to have been inserted into the transposase coding region of a copy of the Tc1 transposon-like element recently identified in salmonids. Received: 9 October 1996 / Accepted: 20 May 1997     

Significant Deviations in the Configurations of Homologous Tandem Repeats in Prokaryotic Genomes

We explored the possibilities of whole-genome duplication (WGD) in prokaryotic species,where we performed statistical analyses of the configurations of the central angles between homologous tandem repeats (TRs) on the circular chromosomes.At first,we detected TRs on their chromosomes and identified equivalent tandem repeat pairs (ETRPs); here,an ETRP is defined as a pair of tandem repeats sequentially similar to each other.Then we carried out statistical analyses of the central angle distributions of the de...     

Reticulate Evolution of Parthenospecies of the Lacertidae Rock Lizards: Inheritance of CLsat Tandem Repeats and Anonymous RAPD Markers

The genetic relatedness of several bisexual and of four unisexual Lacerta saxicola complex lizards was studied, using monomer sequences of the complex-specific CLsat tandem repeats and anonymous RAPD markers. Genomes of parthenospecies were shown to include different satellite monomers. The structure of each such monomer is specific for a certain pair of bisexual species. This fact might be interpreted in favor of co-dominant inheritance of these markers in bisexual species hybridogenesis. This idea is supported by the results obtained with RAPD markers; i.e., unisexual species genomes include only the loci characteristic of certain bisexual species. At the same time, in neither case parthenospecies possess specific, autoapomorphic loci that were not present in this or that bisexual species.     

四核苷酸重复序列单链扩增特性及机理探究

简单重复序列广泛存在于多种生物基因组中,其生物学意义越来越受到人们的重视.许多简单重复序列易于扩增变长,某些重复序列的异常延伸是造成一些遗传疾病的直接原因.本研究以20 nt的60种四重复和6种二重复序列单链为模板,系统研究了它们在嗜热DNA聚合酶作用下等温扩增的特点.电泳结果显示,多数单链模板能扩增变长,即使链内没有互补碱基的序列也可被扩增,如(AGGA)5.定量分析结果显示:回文序列扩增最快;二重复序列比相同碱基组成的四重复序列有更宽的适于扩增的温度范围;G和C含量多的DNA较G和C含量少的序列更易扩增,而且G和C含量越多越适于在较高的温度下扩增;重复单位含两相同嘧啶的链多数比其互补链更易扩增;产物浓度与时间基本呈线性关系.限制性酶切产物结果显示,扩增产物与模板具有相同的重复单位,是重复序列的简单延伸.最后,根据实验结果和相关文献,提出了包括链内滑动扩增和发卡DNA介导扩增两阶段的重复序列单链扩增模型,以对重复序列非特异扩增和相关疾病发生机制的研究提供参考.     

Comparative Genetics of Functional Trinucleotide Tandem Repeats in Humans and Apes

Several human neurodegenerative disorders are caused by the expansion of polymorphic trinucleotide repeat regions. Many of these loci are functional short tandem repeats (STRs) located in brain-expressed genes, and their study is thus relevant from both a medical and an evolutionary point of view. The aims of our study are to infer the comparative pattern of variation and evolution of this set of loci in order to show species-specific features in this group of STRs and on their potential for expansion (therefore, an insight into evolutionary medicine) and to unravel whether any human-specific feature may be identified in brain-expressed genes involved in human disease. We analyzed the variability of the normal range of seven expanding STR CAG/CTG loci (SCA1, SCA2, SCA3-MJD, SCA6, SCA8, SCA12, and DRPLA) and two nonexpanding polymorphic CAG loci (KCNN3 and NCOA3) in humans, chimpanzees, gorillas, and orangutans. The study showed a general conservation of the repetitive tract and of the polymorphism in the four species and high heterogeneity among loci distributions. Humans present slightly larger alleles than the rest of species but a more relevant difference appears in variability levels: Humans are the species with the largest variance, although only for the expanding loci, suggesting a relationship between variability levels and expansion potential. The sequence analysis shows high levels of sequence conservation among species, a lack of correspondence between interruption patterns and variability levels, and signs of conservative selective pressure for some of the STR loci. Only two loci (SCA1 and SCA8) show a human specific distribution, with larger alleles than the rest of species. This could account, at the same time, for a human-specific trait and a predisposition to disease through expansion.This article contains online supplementary material.