甘蓝型油菜-新疆野生油菜二体异附加系BAC文库的构建

Construction of the BAC library of Brassica napus-Chinese Xinjiang wild rape(Sinapis arvensis) disomic alien addition line

甘蓝型油菜-新疆野生油菜二体异附加系包含了甘蓝型油菜全基因组和新疆野生油菜一对染色体,携带有恢复基因;以该二体异附加系为材料,以CopyControlTM pCC1BACTM(HindⅢCloning-Ready)Vector为载体,构建了其基因组的BAC文库(bacterial artificial chromosome library)。该文库含有69 120个克隆,平均插入片段达110kb,相当于甘蓝型油菜单倍体基因组大小的6.336倍。随机挑选3个大于100kb的克隆进行稳定性继代实验,经100代后插入的DNA片段仍然稳定存在。该材料BAC文库的建成,为我们下一步开展甘蓝型油菜基因组学研究及相关重要功能基因的克隆等提供了必要的技术平台。

Brassica napus-Xinjiang wild rape Sinapis arvensis disomic alien addition line, carrying restoring gene, has a complete chromosome set of Brassica napus and a chromosome pair of Sinapis arvensis, its bacterial artificial chromosome （BAC） library has been constructed with CopyControl^TM pCC1 BAC^TM （Hind Ⅲ Cloning -Ready） Vector. This BAC library consists of 69 120 clones. From the analysis of randomly selected BAC clones, the average insert is estimated about 110kb. It is reported that the Brassica napus haploid genome size is 1.2×10^9bp, the library represents about 6.336 x haploid genome equivalents. The insert is stable in a hundred generations, which provides a 99.82% probability of finding any specific genes. The results indicate that this BAC library has high quality with high coverage, it is sufficient for the target gene isolation as well as for the research of Brassica napus and Xinjiang wild rape genomes.