云南紫稻细胞质雄性不育系和保持系线粒体基因组BAC文库的构建

BAC Library Construction for Mitochondrial Genome of Sterile and Maintainer Lines from Yunnan Purple Rice

以云南紫稻细胞质无花粉型水稻樱香不育系和保持系为材料，构建了不育系和保持系的线粒体基因组BAC文库．每个文库均保存2304个菌落，外源插入片段介于10～20kb之间．将两个文库的2304个克隆分别集中于96孔板中，根据已发表的水稻线粒体序列对atp6、atp9、cob、cox2等4个基因设计特定引物，对96孔板中的菌落进行了PCR鉴定，均能筛选得到阳性克隆，通过对96孔板中阳性克隆的位置进行推导，可以得到文库中含有相应基因克隆的大致位置及数目．PCR筛选结果显示两个文库均包含用于检测的目的线粒体基因，说明文库典型性良好，并且PCR法用于文库基因筛选具有快速简便的优点．

Using the sterile line and maintainer line of Yunnan purple rice as materials, we have constructed the bacterial artificial chromosome （BAC） libraries of their mitochondrial genome. Each library consisted of 2 304 clones and the insert size range from 10-20 kb. The libraries were screened by PCR analysis using oligonucleotide primers specific for the desired gene （atp6, atp9, cob, cox2）. The PCR results showed that the libraries can be used for the mitochondrial genome research and the PCR method is suitable for screening the BAC libraries.