大白菜细菌人工染色体文库的构建及鉴定

Construction and Characterization of a Bacterial Artificial Chromosome Library from Chinese Cabbage

以我国优良的大白菜自交系‘85-1’为材料,利用pIndigoBAC-5为载体,通过对高分子量DNA的制备、大片段DNA的选择、连接转化条件等几个方面的优化,构建了大白菜细菌人工染色体文库。该文库由57600个克隆组成,平均大小为98.4kb,空载率为1.5%;覆盖大白菜基因组10.3倍;挑取6个克隆培养5d后,经HindⅢ完全酶切检测,其指纹图谱稳定一致。大白菜细菌人工染色体文库的构建为重要功能基因的克隆和定位及比较基因组研究奠定了基础。

A bacterial artificial chromosome library of Brassica campestris L.ssp.pekinensis（Lour.） Olsson（Chinese cabbage）was constructed from inbred line＇85-1＇with the vector pIndigoBAC-5.The key processes of the construction,such as preparation of high molecular weight DNA,selection of digested fragments,condition of ligation and transformation,were studied.The library consists of 57 600 clones in which the average insert size is about 98.4 kb and the empty clones are about 1.5%.The library represents an equivalent of 10.3 fold size of Chinese cabbage genome.Six clones randomly picked from this library show no HindⅢ fingerprint changes after 5 days＇ successive culture,which indicates that the clones in the library are stable.The library will lay the foundation for gene clone,location and comparative genomics research of Brassica.