河豚鱼基因组细菌人工染色体文库的构建  

Construction of a Pufferfish Bacterial Artificial Chromosome Library

作　　者：刘山鸣[1] 刘勇[1] 张小晓 周巍 杨立[1] 张秀清[1] 吴东颖[1] 蔡胜和[1] 高春生[1] 郞铁明 丁立[1] 戴理[1] 刘国仰[1] 杨焕明[1] 

机构地区：[1]中国医学科学院,中国协和医科大学,基础医学研究所,北京100005

出　　处：《中国医学科学院学报》1998年第3期161-167,共7页Acta Academiae Medicinae Sinicae

基　　金："863"国家高科技研究发展计划(102100402)

摘　　要：目的研究模式生物——河豚鱼的基因组以及利用河豚鱼基因组"小而全"的特点研究人类基因组。方法以红鳍东方豚(Fugu rubripes)的精子DNA为材料,以pBelo BAC 11为载体,构建河豚鱼基因组的细菌人工染色体(bacterial artifi-cial chromosome,BAC)格式化文库(gridded library)。结果该文库包含23 040个克隆,分别保存在240块96孔细胞培养板中。经对100个随机抽样的克隆分析表明,插入片段平均大小为100 kb,因此,该文库相当于河豚鱼单倍体基因组(400Mb)的6倍,即6个库容量。酶切分析10个克隆证明,经100代繁殖其插入片段仍然稳定不变。结论该文库符合BAC基因组文库的所有要求。Objective Study of human genome by taking advantage of investigating pufferfish's (Fugu rubripes), compact model vertebrate genome. Methods Construct a pufferfish bacterial artificial chromosome (BAC) library using Japanese pufferfish (Fugu rubripes) sperm DNA and pBelo BACll vector. Results The library consists of 23 040 clones, which have been arrayed in 240 96-well microtiter plates. 100 BAC clones have been selected at random and analyzed for estimation of the average insert size, which turns out to be 100 kb indicating its nearly 6-fold coverage of pufferfish haploid genome. Restrict analysis of 10 BAC clones show that these inserts are stable even propagated for at least 100 cell generations. Conclusions The library accords with the demands of a BAC library.

关 键 词：河豚鱼 细菌人工染色体 格式化基因组文库 

分 类 号：Q959.841[生物学—动物学]