禾本科植物β-淀粉酶基因家族分子进化及响应非生物胁迫的表达模式分析

 β-淀粉酶(beta-amylase,BAM)是一类关键的淀粉水解酶,在禾谷类作物生长发育过程中起着重要作用,与植物多种非生物胁迫响应相关.本研究通过系统发育分析,将水稻、玉米、高粱、谷子、二穗短柄草5 种禾本科植物中共54 个BAM 基因分为10 个同源基因簇,每个同源基因簇都涵盖了这5 个物种,因此推测在禾本科祖先物种中至少含有10 个BAM 基因,并且在禾本科植物分化后没有发生明显的基因丢失事件.基于对编码蛋白质序列的功能分化分析,表明同源基因簇间存在明显的进化速率的差异.对10 个同源基因簇进行了适应性进化检测,发现有3 个同源簇在禾本科植物的进化过程中经历了适应性进化.此外,对水稻β-淀粉酶的表达分析发现,一些β-淀粉酶具有组织特异性表达特征,并且至少有5 个水稻的β-淀粉酶基因具有受到非生物逆境的胁迫而表现出不同的表达模式.本研究结果为进一步探讨禾本科BAM 基因的生物学功能提供了一定的理论基础.     

大黄鱼转录组水平miRNA的生物信息学分析

根据成熟miRNA序列在物种间高度保守的特性,miRNA前体可以形成茎环状的二级结构,以及miRNA非编码的特性,搭建出了miRNA挖掘的生物信息学流程,并利用此流程在大黄鱼全转录组中进行miRNA的挖掘,得到54条成熟miRNA序列以及54条前体序列(pre-miRNA),根据其种子区的序列将其归为29个miRNA家族。通过对预测得到的miRNA前体序列同其他物种中相同家族的miRNA前体序列构建系统进化树,揭示出同一家族的miRNA前体在不同物种之间的保守性和多样性。利用两种靶基因预测软件,对成熟miRNA进行靶基因预测,共得到2360个靶基因,并对这些靶基因使用BLAST2GO做了进一步的GO注释和富集分析。     

DNA序列碱基组合的频率矩阵及其应用

基于碱基组合在DNA序列中出现的频率,构造11个物种的β-globin基因第一个外显子的编码序列的频率矩阵.借助矩阵2-范数对11个物种进行相似性比较,并结合柱状图对物种之间的相似性进行分析.研究结果表明:所构造的DNA序列频率矩阵不仅能够反映出DNA序列中碱基及碱基组合的含量分布,而且能够显示出序列碱基突变的情况.     

miRNAs在不同物种进化中序列保守性的研究

miRNAs在不同物种的进化中存在保守性，不仅单独在动物或是植物中存在保守性，而且在动植物之间也同样存在．本研究在人和动物的不同组织中克隆到了同源性很高的序列，将所得结果与已发表的成熟的miRNAs比对，发现克隆到的序列与植物中miRNA319家族的相似性较高，并且保守区靠近其5’端，推测新miRNAs可能与miRNA319扮演相似的角色，而这些序列的保守性跨越不同进化阶段的物种，也为miRNAs广泛的保守性增加新的证据，显示miRNAs在物种进化过程中发挥着重要的功能．     

通过同源搜索分析线粒体蛋白转运系统的进化

基于酵母线粒体蛋白转运系统的35个亚基,作者通过同源查找的方法,在27个不同进化层次物种的蛋白组和基因组序列中搜索线粒体蛋白转运系统亚基的同源序列,利用序列之间的同源关系进而构建线粒体蛋白转运系统的进化史.结果显示,线粒体蛋白转运系统的35个亚基中有6个可以在原核物种中找到同源序列,显示了它们的原核起源;线粒体蛋白转运系统的核心亚基出现在真核生物进化早期;其他亚基在真核物种的不同进化分枝中表现出了多样性,并且可以看到一些物种特异性亚基.     

miR-1/133基因簇的进化及其调控网络

miR-1/133基因簇在心肌和骨骼肌中特异性表达,对心脏以及骨骼肌的发育及生理病理具有重要作用.该研究采用生物信息学方法,研究了miR-1/133基因簇的进化特征,预测了其靶基因及其调控的生物学过程.利用miRBase数据库对23种不同物种间miR-1/133基因簇的序列、组织方式进行了比较.采用最大似然法构建miR-1/133基因簇的系统进化树,显示该基因簇的进化与物种进化关系有一定差异.利用在线数据库对miR-1/133基因簇上游转录因子及下游靶基因进行预测,并对其进行综合分析,绘制出该基因簇的网络调控图,表明miR-1/133基因簇参与了骨骼肌与心肌发育、胰岛素受体信号通路、经典Wnt信号通路、p53信号通路等体内重要生理过程,为进一步阐明miR-1/133基因簇的生物学功能提供了理论依据.     

美洲鲎miRNA的生物信息学挖掘与分析

利用miRBase数据库中已有动物的小RNA(miRNA),通过生物信息学方法对美洲鲎(Limulus polyphemus)表达序列标签(Expressed Sequences Tag,EST)和c DNA进行序列比对,挖掘美洲鲎miRNA,得到了72个miRNA前体(pre-miRNA)、49个可编码成熟miRNA,隶属于40个miRNA家族。miRNA家族归类结果表明,无论在低等脊椎动物还是高等动物中,miRNA家族的存在都是相当保守的。以miR-10基因前体为例,分析其序列与其他物种同源序列的差异,显示miRNA序列的高度保守性。miR-10前体序列分析系统进化发现,物种聚类与传统分类亲缘关系一致。     

被子植物中核糖体失活蛋白基因家族分子进化研究

利用34种具有全基因组数据的代表植物,通过生物信息分析方法,深入挖掘和分析不同植物类群基因组中RIPs基因家族的成员构成,推测其可能的扩增方式和功能分化,并探讨了该基因家族在植物中的总体进化趋势.采用BLAST和HMMsearch两种检索方法共鉴定出79个RIPs基因家族成员;通过序列比对和系统进化树的构建,发现RIPs在单双子叶中存在双起源,RIPs在两者分化之前就已存在.它们可能为适应复杂的环境而出现在早期陆生植物中,随后在长期进化过程中不断发生谱系的扩张和拷贝丢失,最后通过功能分化在不同植物中保留下来,其在被子植物进化的过程中有物种特异性的重复.     

14种帘蛤科贝类线粒体基因组特征与系统进化分析

利用从genbank数据库摘录了帘蛤科Veneridae的14种贝类的线粒体基因组全序列,分析其特征,并研究帘蛤科贝类分子系统进化关系。结果显示,A+T碱基含量高于G+C碱基含量,线粒体基因组表现出明显的碱基偏倚现象。对蛋白质编码基因分析,ATG并不是唯一的起始密码子,终止密码子有TAA和TAG两种;密码子组成表现出亲缘关系近的密码子组成差异较小,而亲缘关系远的差异较大。研究还发现14个物种线粒体基因排列顺序存在差异,有基因重排现象和ATP8基因缺失现象,可能是和物种生活环境有关。选取巴非蛤属的和蔼巴非蛤,真曲巴非蛤,织锦巴非蛤,波纹巴非蛤来研究帘蛤科贝类线粒体基因组选择压力,结果表明其12种蛋白质编码基因的Ka/Ks均介于0和1之间,表明基因受到纯化选择作用。基于线粒体基因组序列计算的遗传距离和构建的NJ系统发育树所得结论与传统形态分类基本一致,说明线粒体基因适合作为帘蛤科系统发育的研究手段。     

食肉目核糖核酸酶10分子进化研究

基于食肉目14个物种开展核糖核酸酶10RNase10分子进化研究,获得14条(RNase10)序列,氨基酸序列具有8个结构半光氨酸,但CKXXNTF功能区及催化活性氨基酸发生了改变,等电点明显低于核糖核酸酶基因(RNASE A)超家族典型成员RNase1-8,揭示食肉目RNase10可能失去了RNASE A超家族原有的宿主防御功能,而具有其他新功能.另外,基于RNase10构建的系统发育树为食肉目亲缘关系提供了一种可能性参考.总之,基于食肉目RNase10开展的分子进化研究,增加了该基因研究的多样性,为后续深入研究奠定了基础.     

Minimal ProtoHox cluster inferred from bilaterian and cnidarian Hox complements

Bilaterian animals have a Hox gene cluster essential for patterning the main body axis, and a ParaHox gene cluster. Comparison of Hox and ParaHox genes has led workers to postulate that both clusters originated from the duplication of an ancient cluster named ProtoHox, which contained up to four genes with at least the precursors of anterior and posterior Hox/ParaHox genes. However, the way in which genes diversified within the ProtoHox, Hox and ParaHox clusters remains unclear because no systematic study of non-bilaterian animals exists. Here we characterize the full Hox/ParaHox gene complements and genomic organization in two cnidarian species (Nematostella vectensis and Hydra magnipapillata), and suggest a ProtoHox cluster simpler than originally thought on the basis of three arguments. First, both species possess bilaterian-like anterior Hox genes, but their non-anterior genes do not appear as counterparts of either bilaterian central or posterior genes; second, two clustered ParaHox genes, Gsx and a gene related to Xlox and Cdx, are found in Nematostella vectensis; and third, we do not find clear phylogenetic support for a common origin of bilaterian Cdx and posterior genes, which might therefore have appeared after the ProtoHox cluster duplication. Consequently, the ProtoHox cluster might have consisted of only two anterior genes. Non-anterior genes could have appeared independently in the Hox and ParaHox clusters, possibly after the separation of bilaterians and cnidarians.     

Positive darwinian selection at the imprinted MEDEA locus in plants

In mammals and seed plants, a subset of genes is regulated by genomic imprinting where an allele's activity depends on its parental origin. The parental conflict theory suggests that genomic imprinting evolved after the emergence of an embryo-nourishing tissue (placenta and endosperm), resulting in an intragenomic parental conflict over the allocation of nutrients from mother to offspring. It was predicted that imprinted genes, which arose through antagonistic co-evolution driven by a parental conflict, should be subject to positive darwinian selection. Here we show that the imprinted plant gene MEDEA (MEA), which is essential for seed development, originated during a whole-genome duplication 35 to 85 million years ago. After duplication, MEA underwent positive darwinian selection consistent with neo-functionalization and the parental conflict theory. MEA continues to evolve rapidly in the out-crossing species Arabidopsis lyrata but not in the self-fertilizing species Arabidopsis thaliana, where parental conflicts are reduced. The paralogue of MEA, SWINGER (SWN; also called EZA1), is not imprinted and evolved under strong purifying selection because it probably retained the ancestral function of the common precursor gene. The evolution of MEA suggests a late origin of genomic imprinting within the Brassicaceae, whereas imprinting is thought to have originated early within the mammalian lineage.     

Proof and evolutionary analysis of ancient genome duplication in the yeast Saccharomyces cerevisiae

Whole-genome duplication followed by massive gene loss and specialization has long been postulated as a powerful mechanism of evolutionary innovation. Recently, it has become possible to test this notion by searching complete genome sequence for signs of ancient duplication. Here, we show that the yeast Saccharomyces cerevisiae arose from ancient whole-genome duplication, by sequencing and analysing Kluyveromyces waltii, a related yeast species that diverged before the duplication. The two genomes are related by a 1:2 mapping, with each region of K. waltii corresponding to two regions of S. cerevisiae, as expected for whole-genome duplication. This resolves the long-standing controversy on the ancestry of the yeast genome, and makes it possible to study the fate of duplicated genes directly. Strikingly, 95% of cases of accelerated evolution involve only one member of a gene pair, providing strong support for a specific model of evolution, and allowing us to distinguish ancestral and derived functions.     

The grapevine genome sequence suggests ancestral hexaploidization in major angiosperm phyla

The analysis of the first plant genomes provided unexpected evidence for genome duplication events in species that had previously been considered as true diploids on the basis of their genetics. These polyploidization events may have had important consequences in plant evolution, in particular for species radiation and adaptation and for the modulation of functional capacities. Here we report a high-quality draft of the genome sequence of grapevine (Vitis vinifera) obtained from a highly homozygous genotype. The draft sequence of the grapevine genome is the fourth one produced so far for flowering plants, the second for a woody species and the first for a fruit crop (cultivated for both fruit and beverage). Grapevine was selected because of its important place in the cultural heritage of humanity beginning during the Neolithic period. Several large expansions of gene families with roles in aromatic features are observed. The grapevine genome has not undergone recent genome duplication, thus enabling the discovery of ancestral traits and features of the genetic organization of flowering plants. This analysis reveals the contribution of three ancestral genomes to the grapevine haploid content. This ancestral arrangement is common to many dicotyledonous plants but is absent from the genome of rice, which is a monocotyledon. Furthermore, we explain the chronology of previously described whole-genome duplication events in the evolution of flowering plants.     

The medaka draft genome and insights into vertebrate genome evolution

Teleosts comprise more than half of all vertebrate species and have adapted to a variety of marine and freshwater habitats. Their genome evolution and diversification are important subjects for the understanding of vertebrate evolution. Although draft genome sequences of two pufferfishes have been published, analysis of more fish genomes is desirable. Here we report a high-quality draft genome sequence of a small egg-laying freshwater teleost, medaka (Oryzias latipes). Medaka is native to East Asia and an excellent model system for a wide range of biology, including ecotoxicology, carcinogenesis, sex determination and developmental genetics. In the assembled medaka genome (700 megabases), which is less than half of the zebrafish genome, we predicted 20,141 genes, including approximately 2,900 new genes, using 5'-end serial analysis of gene expression tag information. We found single nucleotide polymorphisms (SNPs) at an average rate of 3.42% between the two inbred strains derived from two regional populations; this is the highest SNP rate seen in any vertebrate species. Analyses based on the dense SNP information show a strict genetic separation of 4 million years (Myr) between the two populations, and suggest that differential selective pressures acted on specific gene categories. Four-way comparisons with the human, pufferfish (Tetraodon), zebrafish and medaka genomes revealed that eight major interchromosomal rearrangements took place in a remarkably short period of approximately 50 Myr after the whole-genome duplication event in the teleost ancestor and afterwards, intriguingly, the medaka genome preserved its ancestral karyotype for more than 300 Myr.     

Silencing of microRNAs in vivo with 'antagomirs'

MicroRNAs (miRNAs) are an abundant class of non-coding RNAs that are believed to be important in many biological processes through regulation of gene expression. The precise molecular function of miRNAs in mammals is largely unknown and a better understanding will require loss-of-function studies in vivo. Here we show that a novel class of chemically engineered oligonucleotides, termed 'antagomirs', are efficient and specific silencers of endogenous miRNAs in mice. Intravenous administration of antagomirs against miR-16, miR-122, miR-192 and miR-194 resulted in a marked reduction of corresponding miRNA levels in liver, lung, kidney, heart, intestine, fat, skin, bone marrow, muscle, ovaries and adrenals. The silencing of endogenous miRNAs by this novel method is specific, efficient and long-lasting. The biological significance of silencing miRNAs with the use of antagomirs was studied for miR-122, an abundant liver-specific miRNA. Gene expression and bioinformatic analysis of messenger RNA from antagomir-treated animals revealed that the 3' untranslated regions of upregulated genes are strongly enriched in miR-122 recognition motifs, whereas downregulated genes are depleted in these motifs. Analysis of the functional annotation of downregulated genes specifically predicted that cholesterol biosynthesis genes would be affected by miR-122, and plasma cholesterol measurements showed reduced levels in antagomir-122-treated mice. Our findings show that antagomirs are powerful tools to silence specific miRNAs in vivo and may represent a therapeutic strategy for silencing miRNAs in disease.     

Positive selection of a gene family during the emergence of humans and African apes

Gene duplication followed by adaptive evolution is one of the primary forces for the emergence of new gene function. Here we describe the recent proliferation, transposition and selection of a 20-kilobase (kb) duplicated segment throughout 15 Mb of the short arm of human chromosome 16. The dispersal of this segment was accompanied by considerable variation in chromosomal-map location and copy number among hominoid species. In humans, we identified a gene family (morpheus) within the duplicated segment. Comparison of putative protein-encoding exons revealed the most extreme case of positive selection among hominoids. The major episode of enhanced amino-acid replacement occurred after the separation of human and great-ape lineages from the orangutan. Positive selection continued to alter amino-acid composition after the divergence of human and chimpanzee lineages. The rapidity and bias for amino-acid-altering nucleotide changes suggest adaptive evolution of the morpheus gene family during the emergence of humans and African apes. Moreover, some genes emerge and evolve very rapidly, generating copies that bear little similarity to their ancestral precursors. Consequently, a small fraction of human genes may not possess discernible orthologues within the genomes of model organisms.     

Gene duplication plays a major role in gene co-option: Studies into the evolution of the motilin/ghrelin family and their receptors

Extant genes can be modified, or ‘tinkered with’, to provide new roles or new characteristics of these genes. At the genetic level, this often involves gene duplication and specialization of the resulting genes into particular functions. We investigate how ligand-receptor partnerships evolve after gene duplication. While significant work has been conducted in this area, the examination of additional models should help us better understand the proposed models and potentially reveal novel evolutionary pattern...     

真核生物中支架蛋白家族(IQGAP family)的分子进化分析

支架蛋白家族(IQGAP family)广泛存在于真核生物中,在细胞的信号转导、骨架运动、细胞分裂的过程中都有着重要功能.研究分析IQGAP蛋白家族的分子进化,有助于深入全面地了解整个IQGAP蛋白家族在不同物种中功能.通过使用相关数据库,在35个真核生物物种中检索到了70个支架蛋白家族成员的序列信息,并对其进行了系统的分子进化研究.我们发现:支架蛋白家族基因广泛存在于后生动物(Metazoan)、真菌(Fungi)、变形虫界(Amoebozoa)以及其他一些真核生物中,提示支架蛋白存在于最后的真核生物的共同祖先(Last Eukaryotic Common Ancestor,LECA).系统发育分析表明脊椎动物的IQGAP蛋白和非脊椎动物中代表性IQGAP基因共享一个共同的祖先基因,单个的IQGAP祖先基因在早期脊椎动物中(在四足总纲和硬骨鱼纲的分离之前),发生了两次基因复制事件,导致现存的脊椎动物中3组IQGAP基因(IQGAP1,IQGAP2,IQGAP3)产生.