A comparison of whole-genome shotgun-derived mouse chromosome 16 and the human genome

The high degree of similarity between the mouse and human genomes is demonstrated through analysis of the sequence of mouse chromosome 16 (Mmu 16), which was obtained as part of a whole-genome shotgun assembly of the mouse genome. The mouse genome is about 10% smaller than the human genome, owing to a lower repetitive DNA content. Comparison of the structure and protein-coding potential of Mmu 16 with that of the homologous segments of the human genome identifies regions of conserved synteny with human chromosomes (Hsa) 3, 8, 12, 16, 21, and 22. Gene content and order are highly conserved between Mmu 16 and the syntenic blocks of the human genome. Of the 731 predicted genes on Mmu 16, 509 align with orthologs on the corresponding portions of the human genome, 44 are likely paralogous to these genes, and 164 genes have homologs elsewhere in the human genome; there are 14 genes for which we could find no human counterpart.     

Functional genomic analysis of the Wnt-wingless signaling pathway

The Wnt-Wingless (Wg) pathway is one of a core set of evolutionarily conserved signaling pathways that regulates many aspects of metazoan development. Aberrant Wnt signaling has been linked to human disease. In the present study, we used a genomewide RNA interference (RNAi) screen in Drosophila cells to screen for regulators of the Wnt pathway. We identified 238 potential regulators, which include known pathway components, genes with functions not previously linked to this pathway, and genes with no previously assigned functions. Reciprocal-Best-Blast analyses reveal that 50% of the genes identified in the screen have human orthologs, of which approximately 18% are associated with human disease. Functional assays of selected genes from the cell-based screen in Drosophila, mammalian cells, and zebrafish embryos demonstrated that these genes have evolutionarily conserved functions in Wnt signaling. High-throughput RNAi screens in cultured cells, followed by functional analyses in model organisms, prove to be a rapid means of identifying regulators of signaling pathways implicated in development and disease.     

The Xist RNA gene evolved in eutherians by pseudogenization of a protein-coding gene

The Xist noncoding RNA is the key initiator of the process of X chromosome inactivation in eutherian mammals, but its precise function and origin remain unknown. Although Xist is well conserved among eutherians, until now, no homolog has been identified in other mammals. We show here that Xist evolved, at least partly, from a protein-coding gene and that the loss of protein-coding function of the proto-Xist coincides with the four flanking protein genes becoming pseudogenes. This event occurred after the divergence between eutherians and marsupials, which suggests that mechanisms of dosage compensation have evolved independently in both lineages.     

The Release 5.1 annotation of Drosophila melanogaster heterochromatin

The repetitive DNA that constitutes most of the heterochromatic regions of metazoan genomes has hindered the comprehensive analysis of gene content and other functions. We have generated a detailed computational and manual annotation of 24 megabases of heterochromatic sequence in the Release 5 Drosophila melanogaster genome sequence. The heterochromatin contains a minimum of 230 to 254 protein-coding genes, which are conserved in other Drosophilids and more diverged species, as well as 32 pseudogenes and 13 noncoding RNAs. Improved methods revealed that more than 77% of this heterochromatin sequence, including introns and intergenic regions, is composed of fragmented and nested transposable elements and other repeated DNAs. Drosophila heterochromatin contains "islands" of highly conserved genes embedded in these "oceans" of complex repeats, which may require special expression and splicing mechanisms.     

A systematic survey of loss-of-function variants in human protein-coding genes

Genome-sequencing studies indicate that all humans carry many genetic variants predicted to cause loss of function (LoF) of protein-coding genes, suggesting unexpected redundancy in the human genome. Here we apply stringent filters to 2951 putative LoF variants obtained from 185 human genomes to determine their true prevalence and properties. We estimate that human genomes typically contain ~100 genuine LoF variants with ~20 genes completely inactivated. We identify rare and likely deleterious LoF alleles, including 26 known and 21 predicted severe disease-causing variants, as well as common LoF variants in nonessential genes. We describe functional and evolutionary differences between LoF-tolerant and recessive disease genes and a method for using these differences to prioritize candidate genes found in clinical sequencing studies.     

锯缘青蟹Sox基因的PCR扩增(英文)

SRY基因(sex-determining region of the Y chromosome)是人类及哺乳动物睾丸决定因子TDF(testisdetermining factor)的最佳候选基因。SRY蛋白含有204个氨基酸,其中1个约79个氨基酸区域即HMG盒(high mobility group box)是SRY基因编码蛋白质的唯一功能区。由于SRY的发现,人们进而发现了1个庞大的与性别决定有关的SRY盒-Sox(SRY-related HMG box gene)基因家族。锯缘青蟹是我国重要的海洋经济蟹类之一,其性染色体和性别决定机制仍处于进化的早期阶段,在分子水平上探讨其性别决定机制尚不多见。通过采用PCR技术,以特异扩增人SRY基因HMG盒保守区的1对兼并引物,扩增了锯缘青蟹基因组的Sox基因。结果表明锯缘青蟹雌雄个体与人一样,均能扩增出1条大小约为216bp左右的基因片段,显示出该基因在进化上的高度保守性,为探索锯缘青蟹的性别决定机制及Sox基因的进化提供了分子资料。     

锯缘青蟹Sox基因的PCR扩增

SRY基因(sex-determining region of the Y chromosome)是人类及哺乳动物睾丸决定因子TDF(testisdetermining factor)的最佳候选基因。SRY蛋白含有204个氨基酸,其中1个约79个氨基酸区域即HMG盒(high mobility group box)是SRY基因编码蛋白质的唯一功能区。由于SRY的发现,人们进而发现了1个庞大的与性别决定有关的SRY盒-Sox(SRY-related HMG box gene)基因家族。锯缘青蟹是我国重要的海洋经济蟹类之一,其性染色体和性别决定机制仍处于进化的早期阶段,在分子水平上探讨其性别决定机制尚不多见。通过采用PCR技术,以特异扩增人SRY基因HMG盒保守区的1对兼并引物,扩增了锯缘青蟹基因组的Sox基因。结果表明锯缘青蟹雌雄个体与人一样,均能扩增出1条大小约为216bp左右的基因片段,显示出该基因在进化上的高度保守性,为探索锯缘青蟹的性别决定机制及Sox基因的进化提供了分子资料。     

Sequence Comparison of MHC Class Ⅱβ (Exon 2) and Phylogenetic Relationship Between Poultry and Mammalian

A fragment spanning over exon 2 and intron 2 of major histocompatibility complex B-LB Ⅱ genes was amplified using PCR,cloned and sequenced in 13 individuals from eight Chinese indigenous chicken breeds and one introduced breed. Another 41 sequences of MHC class Ⅱβ from ten vertebrate species were cited from the NCBI GenBank. Thirteen new B-LB Ⅱ alleles were found in the chicken breeds sampled. Alignment of the exon 2 sequences revealed 91.1-97.8% similarity to each other within the chickens sampled, and the chickens shared 84.1-87.0% homology to Phasianus colchicus, 78.5-81.5% similarity to Coturnixjaponica. The sequences in poultry showed 62.6-68.1% identity to HLA-DRBl, 50-61.5% similarity to DQB (HLA-, SLA- and H2-BB), 53.7-60% to HLA-DPB and 53.3-57.8% similarity to HLA-DOB. The frequency of nonsynonymous substitutions of nucleotide was higher than that of synonymous substitutions, and the frequencies of nonsynonymous and synonymous substitutions in poultry B-LB Ⅱ genes were lower than those observed in mammalian DRB1 and DQB1 genes. The deduced amino acid sequences of MHC class Ⅱβ1 domain exhibited extreme difference in conversed region and variable region patterns among the various species, but the two conserved cysteines forming disulfide-bond were shown consistent in poultry with that in mammalian species; and the carbohydrate attachment site was found more conserved in chicken, Homo sapiens, Bos taurus, Ovis aries and Capra hircus than in Sus scrofa and rodent animals. Compared with exon 2 of DQB1 genes of Homo sapiens, ruminant species and Sus scrofa, the differentia that the deletion of six nucleotides at position195 to 200 of exon 2 of DQB1 genes, and insertion of three nucleotides at position 247 to 249 of the exon 2 existed in rodent species were found, which led to the absence of three AA residues at position 65, 66,and 67 within β1 domain of DQB1 chain, and the insertion of one AA residue at position 85. The difference of the deletion of six nucleotides at position 72 to 77 of exon 2 of DPB1 genes was observed with Homo sapiens DQB1, which caused absence of three AA residues at position 24, 25, and 26 of β1 domain of DPBl chain. The phylogenetic tree revealed that the B-LB Ⅱ sequences from poultry are not orthologous to the class Ⅱ MHC β-chain genes of mammalian species. The tree indicated that genetic evolutionary relationship of chickens with Phasianus colchicus was much closer than with Coturnixjaponica, and the DQB and DPB clusters are more tightly related to each other than to the remaining clusters.     

The dog genome: survey sequencing and comparative analysis

A survey of the dog genome sequence (6.22 million sequence reads; 1.5x coverage) demonstrates the power of sample sequencing for comparative analysis of mammalian genomes and the generation of species-specific resources. More than 650 million base pairs (>25%) of dog sequence align uniquely to the human genome, including fragments of putative orthologs for 18,473 of 24,567 annotated human genes. Mutation rates, conserved synteny, repeat content, and phylogeny can be compared among human, mouse, and dog. A variety of polymorphic elements are identified that will be valuable for mapping the genetic basis of diseases and traits in the dog.     

Evolution of yeast noncoding RNAs reveals an alternative mechanism for widespread intron loss

The evolutionary forces responsible for intron loss are unresolved. Whereas research has focused on protein-coding genes, here we analyze noncoding small nucleolar RNA (snoRNA) genes in which introns, rather than exons, are typically the functional elements. Within the yeast lineage exemplified by the human pathogen Candida albicans, we find--through deep RNA sequencing and genome-wide annotation of splice junctions--extreme compaction and loss of associated exons, but retention of snoRNAs within introns. In the Saccharomyces yeast lineage, however, we find it is the introns that have been lost through widespread degeneration of splicing signals. This intron loss, perhaps facilitated by innovations in snoRNA processing, is distinct from that observed in protein-coding genes with respect to both mechanism and evolutionary timing.     

Phosphorylation of retinoblastoma protein by viral protein with cyclin-dependent kinase function

As obligate intracellular parasites, viruses expertly modify cellular processes to facilitate their replication and spread, often by encoding genes that mimic the functions of cellular proteins while lacking regulatory features that modify their activity. We show that the human cytomegalovirus UL97 protein has activities similar to cellular cyclin-cyclin-dependent kinase (CDK) complexes. UL97 phosphorylated and inactivated the retinoblastoma tumor suppressor, stimulated cell cycle progression in mammalian cells, and rescued proliferation of Saccharomyces cerevisiae lacking CDK activity. UL97 is not inhibited by the CDK inhibitor p21 and lacks amino acid residues conserved in the CDKs that permit the attenuation of kinase activity. Thus, UL97 represents a functional ortholog of cellular CDKs that is immune from normal CDK control mechanisms.     

Draft genome of the filarial nematode parasite Brugia malayi

Parasitic nematodes that cause elephantiasis and river blindness threaten hundreds of millions of people in the developing world. We have sequenced the approximately 90 megabase (Mb) genome of the human filarial parasite Brugia malayi and predict approximately 11,500 protein coding genes in 71 Mb of robustly assembled sequence. Comparative analysis with the free-living, model nematode Caenorhabditis elegans revealed that, despite these genes having maintained little conservation of local synteny during approximately 350 million years of evolution, they largely remain in linkage on chromosomal units. More than 100 conserved operons were identified. Analysis of the predicted proteome provides evidence for adaptations of B. malayi to niches in its human and vector hosts and insights into the molecular basis of a mutualistic relationship with its Wolbachia endosymbiont. These findings offer a foundation for rational drug design.     

紫貂线粒体基因组全序列结构及其进化

利用PCR方法和直接测序技术获得的紫貂线粒体基因组全长为16523bp,包含13个蛋白质编码基因、2个rRNA基因、22个tRNA基因和1个非编码基因控制区(D-Loop区),碱基组成为A32.0%、C27.6%、G14.7%、T25.8%;在编码蛋白质基因的密码子第3位点处具有AC高偏向性(72.6%),并且频繁利用不完全终止密码子T或TA(7个)。将紫貂线粒体基因组序列提交到GenBank,并获得检索号为FJ429093。结合GenBank中已公布的鼬科其他6种动物的线粒体基因组全序列及貂属6种D-Loop区部分序列,分别以虎和狗獾为外类群,应用最大简约法构建鼬科和貂属物种的系统进化树。结果表明:鼬科7个物种分成3大支系,分别为水獭亚科和臭鼬亚科群、日本貂和貂熊群以及紫貂和狗獾群,说明鼬科并非是一个单系群,紫貂与鼬科中的獾亚科亲缘关系较近,而与貂属中的美洲貂和石貂亲缘关系最近。     

猪蛋白编码基因3'UTR中IRPRE1元件的鉴定及其基因特征分析

为鉴定猪全基因组范围内蛋白编码基因3'UTR(3'–untranslated region)中反向重复PRE1(inverted repeated PRE1,IRPRE1)元件,对猪全基因组的22342个蛋白编码基因的3'UTR序列进行重复序列元件的生物信息学分析。结果表明:猪蛋白编码基因的3'UTR序列中短散在重复序列与简单重复序列在重复序列的类别中所占比例较高,分别为27.58%与31.08%;在SINE/t RNA的重复元件中,Pre0_SS和PRE1x元件所占的比例较高,分别为41.83%和37.51%;共有1 094个候选蛋白编码基因的3'UTR中含有IRPRE1元件;GO分析发现这些候选基因主要参与m RNA经由剪接体的剪接、细胞分裂、RNA通过酯交换反应发生的剪接、T细胞激活、RNA剪接、T细胞受体信号通路、对糖苷反应、甘油三酯稳态、外源性凋亡信号通路的正调节及胆固醇合成等生物过程;KEGG pathway分析发现这些候选基因参与了缬草碱、亮氨酸和异亮氨酸降解途径、TNF信号通路、T细胞受体信号通路、RIG–I样受体信号通路、吞噬体、剪接体、胆汁分泌、甲状腺激素合成和凋亡;对3个蛋白编码基因的3'UTR中的IRPRE1元件进行鉴定发现,其在多个组织中广泛表达。     

Evolution and functional impact of rare coding variation from deep sequencing of human exomes

As a first step toward understanding how rare variants contribute to risk for complex diseases, we sequenced 15,585 human protein-coding genes to an average median depth of 111× in 2440 individuals of European (n = 1351) and African (n = 1088) ancestry. We identified over 500,000 single-nucleotide variants (SNVs), the majority of which were rare (86% with a minor allele frequency less than 0.5%), previously unknown (82%), and population-specific (82%). On average, 2.3% of the 13,595 SNVs each person carried were predicted to affect protein function of ~313 genes per genome, and ~95.7% of SNVs predicted to be functionally important were rare. This excess of rare functional variants is due to the combined effects of explosive, recent accelerated population growth and weak purifying selection. Furthermore, we show that large sample sizes will be required to associate rare variants with complex traits.