白颊长臂猿Fosmid 基因组文库的构建
文献类型：学位论文
| 作者 | 陈丽萍 |
| 学位类别 | 硕士 |
| 答辩日期 | 2007-02 |
| 授予单位 | 中国科学院研究生院 |
| 授予地点 | 北京 |
| 导师 | 杨凤堂 |
| 关键词 | 白颊长臂猿 Fosmid 基因组文库 FISH定位 染色体 |
| 其他题名 | Construction, characterization and chromosomal mapping of a Fosmid library of white-cheeked gibbon |
| 学位专业 | 细胞生物学 |
| 中文摘要 | 长臂猿和大猿、人类同属人猿超科，在许多生物学特性方面具有相似性，但是自与大猿的祖先分歧以来的约两千万年的时间里，长臂猿染色体进化速度快于人猿超科其他物种和其他灵长类物种，出现了高度重排的核型。利用长臂猿染色体特异涂色探针已在人的染色体上探测到至少80个染色体断裂位点（synteny breakpoint），但是由于染色体荧光原位杂交技术的分辨率太低，仅有约5MB，所以迄今无从了解同源断裂位点两侧的序列特征，也就无法揭示长臂猿染色体快速重排的分子机制。 本论文旨在构建一个长臂猿Fosmid文库，以便为揭示长臂猿染色体快速进化的分子机制奠定基础。我选择了核型高度重排，且在长臂猿属中二倍染色体数目最多（2n=52）的代表物种——白颊长臂猿为材料，构建了一个白颊长臂猿 Fosmid 基因组文库。该文库含有192000个克隆，插入片段在30kb-45kb之间，假定白颊长臂猿的基因组大小按3.410ⅹ6kb计算，该文库至少有2.5倍的基因组覆盖率。通过对100个随机挑选的Fosmid 克隆进行末端测序，获得了200个序列标签。将这100个经末端测序的Fosmid 克隆与人的基因组进行BLAST比对以及将这些克隆制备成探针与白颊长臂猿和人的染色体中期分裂相进行荧光原位杂交（FISH），得到了这100个克隆在人基因组上的精确定位以及其中94个克隆在白颊长臂猿基因组上的精确定位,同时在这100个克隆中没有发现嵌合体克隆的存在。该文库以及所定位的克隆将为研究长臂猿以及其他高等灵长类染色体进化分子机制奠定基础，同时也为非人灵长类的比较基因组研究提供一个宝贵的资源。 |
| 英文摘要 | Gibbons (genus Hylobates, family Hylobatidae) constitute a sister group of the great apes (Pongidae) and humans (Hominidae). Despite gibbons diverged from the lineage leading to the great apes at about 18 million years ago, they differ from other primates and most mammals with an exceptionally high rate of chromosomal rearrangement. At least 80 synteny breakpoints on both human and gibbon chromosome are detected so far. Unfortunately the synteny breakpoints defined by fluorescence in-situ hybridization (FISH) on the chromosomes only have a resolution of about 5MB. Identification of underlying genetic mechanisms, which produced such rearrangements, requires breakpoint characterization at the DNA sequence level. But there is no available genomic library for this species that is required to study the mechanism and consequences of karyotypic changes. As a first step forward, I thus decided to construct a Fosmid genomic library of the gibbon. The white-cheeked gibbon (H. leucogenys) is one of subspecies of Nomascu, and has an exceptionally high rate of chromosomal rearrangement. Hence, I choose the white-cheeked gibbon to construct a genomic Fosmid library. Until now, 192,000 independent Fosmid clones have been constructed. Given the white cheeked gibbon has a haploid C value (CV) of the white cheeked gibbon (3.40), the haploid C values (CV) of 3.40, this clone library represents at least 2 times coverage of the white-cheeked gibbon genome. By end-sequencing of 100 randomly selected Fosmid clones, I generated 200 sequence tags for this gibbon. The 100 end- sequenced Fosmid clones were also mapped onto the chromosomes of the white-cheeked gibbon by FISH, no unwanted chimeric clones have been detected. BLAST search against human genome showed a good correlation between the number of hit clones and the chromosomes and the size of the chromosomes, indicating an unbiased chromosomal distribution of the Fosmid library. This library and the mapped Fosmid clones will serve as a resource for subsequen evolutionary breakpoint studies and comparative genomics studies. |
| 语种 | 中文 |
| 公开日期 | 2010-10-14 |
| 源URL | [http://159.226.149.42:8088/handle/152453/6130]  |
| 专题 | 昆明动物研究所_其他 |
| 推荐引用方式 GB/T 7714 | 陈丽萍. 白颊长臂猿Fosmid 基因组文库的构建[D]. 北京. 中国科学院研究生院. 2007. |
入库方式： OAI收割
来源：昆明动物研究所
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