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.     

Evolutionary discrimination of mammalian conserved non-genic sequences (CNGs)

Analysis of the human and mouse genomes identified an abundance of conserved non-genic sequences (CNGs). The significance and evolutionary depth of their conservation remain unanswered. We have quantified levels and patterns of conservation of 191 CNGs of human chromosome 21 in 14 mammalian species. We found that CNGs are significantly more conserved than protein-coding genes and noncoding RNAS (ncRNAs) within the mammalian class from primates to monotremes to marsupials. The pattern of substitutions in CNGs differed from that seen in protein-coding and ncRNA genes and resembled that of protein-binding regions. About 0.3% to 1% of the human genome corresponds to a previously unknown class of extremely constrained CNGs shared among mammals.     

High resolution of mouse chromosomes: banding conservation between man and mouse

A detailed schematic representation of high-resolution G-banding patterns was prepared from elongated and finely banded mitotic chromosomes of the mouse. Such chromosomes can be obtained from both animal tissue and cell lines by a simple protocol, facilitating precise demarcation of breakpoints in chromosome rearrangements and aiding in the sublocalization of genes. Regions of subbanding homology were observed between human and mouse chromosomal segments known to have conserved gene assignments, an indication that, at the cytogenetic level, extensive regions of the mammalian genome may remain intact after 60 million years of species divergence.     

Assignment of the murine interferon sensitivity and cytoplasmic superoxide dismutase genes to chromosome 16

Both hybrids of mouse and human microcells and whole cell hybrids generated by the fusion of primary mouse cells and SV40-transformed human fibroblasts were used to establish the syntenic association of the murine cytoplasmic superoxide dismutase and the interferon sensitivity genes on mouse chromosome 16. This assignment adds two new markers to chromosome 16 and provides another example of an evolutionarily conserved linkage. This finding also provides an animal model both for cellular responsiveness to interferon and for Down's syndrome.     

Metagenomic analysis of the human distal gut microbiome

The human intestinal microbiota is composed of 10(13) to 10(14) microorganisms whose collective genome ("microbiome") contains at least 100 times as many genes as our own genome. We analyzed approximately 78 million base pairs of unique DNA sequence and 2062 polymerase chain reaction-amplified 16S ribosomal DNA sequences obtained from the fecal DNAs of two healthy adults. Using metabolic function analyses of identified genes, we compared our human genome with the average content of previously sequenced microbial genomes. Our microbiome has significantly enriched metabolism of glycans, amino acids, and xenobiotics; methanogenesis; and 2-methyl-d-erythritol 4-phosphate pathway-mediated biosynthesis of vitamins and isoprenoids. Thus, humans are superorganisms whose metabolism represents an amalgamation of microbial and human attributes.     

A mouse model of the aniridia-Wilms tumor deletion syndrome

Deletion of chromosome 11p13 in humans produces the WAGR syndrome, consisting of aniridia (an absence or malformation of the iris), Wilms tumor (nephroblastoma), genitourinary malformations, and mental retardation. An interspecies backcross between Mus musculus/domesticus and Mus spretus was made in order to map the homologous chromosomal region in the mouse genome and to define an animal model of this syndrome. Nine evolutionarily conserved DNA clones from proximal human 11p were localized on mouse chromosome 2 near Small-eyes (Sey), a semidominant mutation that is phenotypically similar to aniridia. Analysis of Dickie's Small-eye (SeyDey), a poorly viable allele that has pleiotropic effects, revealed the deletion of three clones, f3, f8, and k13, which encompass the aniridia (AN2) and Wilms tumor susceptibility genes in man. Unlike their human counterparts, SeyDey/+ mice do not develop nephroblastomas. These findings suggest that the Small-eye defect is genetically equivalent to human aniridia, but that loss of the murine homolog of the Wilms tumor gene is not sufficient for tumor initiation. A comparison among Sey alleles suggests that the AN2 gene product is required for induction of the lens and nasal placodes.     

Human genome ultraconserved elements are ultraselected

Ultraconserved elements in the human genome are defined as stretches of at least 200 base pairs of DNA that match identically with corresponding regions in the mouse and rat genomes. Most ultraconserved elements are noncoding and have been evolutionarily conserved since mammal and bird ancestors diverged over 300 million years ago. The reason for this extreme conservation remains a mystery. It has been speculated that they are mutational cold spots or regions where every site is under weak but still detectable negative selection. However, analysis of the derived allele frequency spectrum shows that these regions are in fact under negative selection that is much stronger than that in protein coding genes.     

The protein kinase complement of the human genome

We have catalogued the protein kinase complement of the human genome (the "kinome") using public and proprietary genomic, complementary DNA, and expressed sequence tag (EST) sequences. This provides a starting point for comprehensive analysis of protein phosphorylation in normal and disease states, as well as a detailed view of the current state of human genome analysis through a focus on one large gene family. We identify 518 putative protein kinase genes, of which 71 have not previously been reported or described as kinases, and we extend or correct the protein sequences of 56 more kinases. New genes include members of well-studied families as well as previously unidentified families, some of which are conserved in model organisms. Classification and comparison with model organism kinomes identified orthologous groups and highlighted expansions specific to human and other lineages. We also identified 106 protein kinase pseudogenes. Chromosomal mapping revealed several small clusters of kinase genes and revealed that 244 kinases map to disease loci or cancer amplicons.     

A MicroRNA Catalog of Swine Umbilical Vein Endothelial Cells Identified by Deep Sequencing

MicroRNAs (miRNAs) are endogenous ∼22 nt RNAs that play important regulatory roles in targeting mRNAs for cleavage or translational repression. Despite the discovery of increasing numbers of human and mouse miRNAs, little is known about miRNAs from pig. In this study, we sought to extend the repertoire of porcine small regulatory RNAs using Solexa sequencing. We sequenced a library of small RNAs prepared from immortalized swine umbilical vein endothelial cells (SUVECs). We produced over 13.6 million short sequence reads, of which 8547658 perfectly mapped to the pig genome. A bioinformatics pipeline was used to identify authentic mature miRNA sequences. We identified 154 porcine miRNA genes, among which 146 were conserved across species, and 8 were pig-specific miRNA genes. The 146 miRNA genes encoded 116 conserved mature miRNAs and 66 miRNA*. The 8 pig-specific miRNA genes encoded 4 mature miRNAs. Four potential novel miRNAs were identified. The secondary structures of the 154 miRNA genes were predicted; 13 miRNAs have 2 structures, and miR-9 and miR-199 have 4 and 3 structures, respectively. 36 miRNAs were organized into 19 compact clusters. miR-206, miR-21 and miR-378 were the relatively highly expressed miRNAs. In conclusion, Solexa sequencing allowed the successful discovery of known and novel porcine miRNAs with high accuracy and efficiency. Furthermore, our results supply new data to the somewhat insufficient pig miRBase, and are useful for investigating features of the blood-brain barrier, vascular diseases and inflammation.     

Identification of the protein encoded by the E6 transforming gene of bovine papillomavirus

Papillomaviruses (PV) contain several conserved genes that may encode nonstructural proteins; however, none of these predicted gene products have been identified. Papillomavirus E6 genes are retained and expressed as RNA in PV-associated human and animal carcinomas and cell lines. This suggests that the E6 gene product may be important in the maintenance of the malignant phenotype. The E6 open reading frame of the bovine papillomavirus (BPV) genome has been identified as one of two BPV genes that can independently transform mouse cells in vitro. A polypeptide encoded by this region of BPV was produced in a bacterial expression vector and used to raise antisera. The antisera specifically immunoprecipitated the predicted 15.5-kilodalton BPV E6 protein from cells transformed by the E6 gene. The E6 protein was identified in both the nuclear and membrane fractions of these transformed cells.     

桑树WRKY转录因子的全基因组鉴定及生物信息学分析

[目的]明确桑树基因组中WRKY转录因子家族结构及其功能特征,为进一步揭示WRKY转录因子家族生物学功能提供科学依据.[方法]利用生物信息学方法对桑树WRKY转录因子的数目、类型、结构、系统进化关系、保守结构域和密码子使用偏性等进行全面分析.[结果]基于桑树全基因组蛋白数据库,共鉴定出55个桑树WRKY转录因子家族基因,占桑树基因总数(29261)的1.88%.桑树WRKY转录因子存在6种内含子数量类型及15种内含子相位类型,其中27个基因含有2个内含子,25个基因的相位类型为2-2型.保守结构域系统进化分析结果显示,桑树WRKY转录因子家族蛋白主要分为三大类(Ⅰ、Ⅱ和Ⅲ),Ⅰ类可分为ⅠN和ⅠC两个亚组,Ⅱ类根据聚类情况又可分为Ⅱa、Ⅱb、Ⅱc、Ⅱd和Ⅱe等5个亚组.桑树WRKY转录因子蛋白保守结构域分析发现有五类Motif的保守性较强,桑树WRKY转录因子蛋白中均包含C端Motif l,Ⅰ类蛋白同时含有N端Motif 3.桑树WRKY转录因子家族基因启动子区富含PBF(C2H2锌指因子)和AHL(拟南芥hook因子)元件.密码子使用偏性分析结果显示,桑树WRKY转录因子家族基因的有效密码子数(ENC)介于48.00~60.00,密码子第3位GC含量(GC3s)介于0.330~0.722,平均亲水性值(Gravy)均为负值;同义密码子相对使用度(RSCU)>1.000的密码子有29个,且以A(6个)或T(11个)结尾较G(4个)或C(8个)结尾的略多.[结论]桑树WRKY转录因子家族包含55个成员,内含子相位类型一致的同组成员可能来源于同一祖先基因,且与基因复制和基因组重排有关;蛋白序列高度保守,在植物抵御环境胁迫过程中发挥作用;基因密码子使用偏性较弱,主要受碱基突变选择压力影响.     

蜜蜂发育相关基因研究

蜜蜂是一种重要的授粉昆虫,也是研究人类疾病及社会行为的模式生物之一。由其基因组测序可知,蜜蜂基因组中(A+T)及CpG含量较高,其昼夜节律、RNAi和DNA甲基化基因更类似脊椎动物。先天免疫、表皮蛋白和味觉受体基因较少,气味受体基因较多。蜜蜂早期发育途径中的一些基因与果蝇的相似,功能却显著不同。本文综述了蜜蜂卵、幼虫、蛹和成虫期发育相关基因及其研究进展。     

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.     

禾谷类作物的比较基因组研究

 水稻是基因组最小的禾谷类作物,饱和遗传连锁图谱的构建,以及在此基础上开展的标记辅助选择和抗病基因克隆,表明水稻基因组研究已经领先于其他禾谷类作物。比较基因组研究表明：小麦、玉米、高粱、谷子和甘蔗的基因组均可由水稻染色体区段重新排列而成，这些区段上DNA标记的排列顺序在各个种之间保留。各种作物基因组大小的差异可能由于各个区段内基因间重复顺序扩增的程度不同所致。根据这些区段在各种作物染色体的排列顺序，有人提出根据水稻染色体区段排列单个原始禾谷类染色体的设想，为深入研究禾谷类作物的进化遗传提出了全新的思路。禾谷类作物基因组之间的共线性有利于在小基因组内克隆大基因组作物的同源基因，使生物技术在作物育种中发挥更大的作用。     

木薯25个WRKY家族转录因子在生物胁迫下的表达分析

根据拟南芥和水稻的WRKY基因和木薯基因组序列,利用生物信息学方法预测木薯Me WRKY转录因子家族成员,并对其进行系统进化关系和保守结构域的分析。通过病原菌接种处理,分析不同Me WRKY转录因子的表达差异。结果显示,木薯共编码25个与抗病相关WRKY蛋白,在病原菌胁迫条件下,其中16个Me WRKY基因的表达受到显著影响,表明这些WRKY蛋白可能参与木薯的防御应答反应。     

Study on the Complete Mitochondrial Genome of Qaidam Cattle(Bos taurus)

Qaidam cattle(Bos taurus) is an important endemic breed in Northwest China, which is mainly distributed in the northwest of Qinghai-Tibet Plateau. It has strong adaptive ability to plateau and swamp conditions, such as cold tolerance and insect resistance. In this study, the first complete mitochondrial genome of Qaidam cattle was reported. The circular double-stranded genome is 16 340 bp in size, and contains 13 protein-coding genes, 22 transfer RNA genes, two ribosomal RNA genes, and a D-loop region. The overall nucleotide composition is 33.4% A, 27.2% T, 26.0% C and 13.4% G, with a total A +T content of60.6%. The gene order and composition are similar to those of other B. taurus breeds. Molecular phylogenetic analysis indicated that Qaidam cattle was split as an independent clade and nested within Asian cattle breeds.