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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。
Based on the excellent inbred line ’85-1 ’of Chinese cabbage and the use of pIndigoBAC-5 as the carrier, we optimized the preparation of high molecular weight DNA, the selection of large DNA and the transformation conditions Chinese cabbage bacterial artificial chromosome library. The library consisted of 57600 clones, with an average size of 98.4kb and an empty vector rate of 1.5%. It covered 10.3 times of the genome of Chinese cabbage. After 6 clones were picked for 5 days, the fingerprints were completely digested with Hind Ⅲ and the fingerprint was consistent. The construction of bacterial artificial chromosome library of Chinese cabbage laid the foundation for cloning and localization of important functional genes and comparative genomics research.