抗稻瘟病水稻BAC文库的构建与鉴定  

Construction and Characterization of a Bacterial Artificial Chromosome Library from Blast Resistance Rice

作　　者：陈琪[1] 邓一文[2] 黄彬彬[1] 孙晓明[1] 张文文[1] 王刚[1] 许政暟[1] 宋任涛[1] 

机构地区：[1]上海大学生命科学学院 [2]中科院上海植物生理生态研究所,上海200032

出　　处：《上海大学学报（自然科学版）》2007年第3期325-330,共6页Journal of Shanghai University:Natural Science Edition

摘　　要：水稻是一种重要的粮食作物,同时也是一种重要的单子叶植物模式.稻瘟病是水稻生长中的一种严重病害,因此分离和克隆新的稻瘟病抗病基因具有重要的应用价值.以一个抗稻瘟病农家种水稻为材料,以plndigoBAC5为载体,构建了细菌人工染色体(BAC)文库.该文库共有90 000个转化子,插入频率99%,插入片段平均长度为105kb,由此推测这个文库覆盖水稻基因组约20倍.利用其中的45 000个转化子建立了4维PCR筛选体系,并且通过4维PCR筛选体系,筛选获得了7个含水稻分蘖基因(MOC1)的阳性克隆.因此该文库是一个高质量、高覆盖率的水稻BAC文库,能有效用于目的基因的分离.Rice blast is a serious disease in production. Therefore the isolation of novel blast resistance gene is important. We constructed a bacterial artificial chromosome （BAC） library from a rice verity with blast resistance with vector plndigoBAC5. The BAC library consisted of about 90 000 clones, 99 % of the which had insertion with an average insertion size of about 105 kb. The BAC library was estimated to have 20 times genome coverage. Half of the library consisting of about 45 000 clones was used to develop a 4-dimensional PCR screening system, through which 7 positive MOCI clones were obtained. The result indicats that this rice BAC library has high quality and high coverage, and is sufficient for target gene isolation.

关 键 词：水稻 稻瘟病 细菌人工染色体文库 4维PCR筛选体系 

分 类 号：Q785[生物学—分子生物学]