云南药用野生稻BIBAC文库混合克隆池制备及筛选  

Preparation and Screening of Mixed-clones Pools from BIBAC Library of Oryza officinallis in Yunnan Province

作　　者：刘艳平[1,2] 程在全[1] 殷富有[1] 余腾琼[1] 张敦宇[1] 郭怡卿[1] 张薇 黄兴奇[1,2] 

机构地区：[1]云南省农业科学院生物技术与种质资源研究所,昆明650223 [2]云南大学生命科学学院,昆明650091 [3]云南省昭通市农科所,昭通657000

出　　处：《生物技术通报》2012年第2期117-120,共4页Biotechnology Bulletin

基　　金：国家自然科学基金项目(30660090;31160067);国家公益性行业(农业)科研专项(201003021);云南省应用基础研究重点项目(2008C04);云南省高端人才引进项目(2010CI120);云南省攻关项目(2010BB004)

摘　　要：在已构建完成的云南药用野生稻BIBAC(binary bacterial artificial chomosome)文库的基础上,将文库制备成一、二、三级混合克隆池,各级混合池的数量分别为3 360、140和14个。根据Xa21抗病基因序列设计1对特异引物,利用4步PCR法对文库混合克隆池进行逐级筛选,初步确定了3个抗病基因阳性克隆。为今后以PCR法高效利用云南药用野生稻BIBAC文库挖掘其优异基因奠定了良好的基础。Based on the BIBAC library of Oryza officinalis Wall.ex Watt.,three grades mixed-clones pools of the BIBAC library were prepared,including the primary,secondary and tertiary.The number of the primary,secondary and tertiary mixed-clones pools was 3 360,140 and 14,respectively,and the 14 tertiary mixed-clones pools contained entire BIBAC clones of the library.One pair of specific primers was designed according to the reported sequence of Xa21 which features broad-spectrum resistance against bacterial blight caused by different strains of Xanthomonas oryzae pv.oryzae（Xoo）.Three positive BIBAC clones containing Xa21 DNA fragment were obtained through a four-step PCR screening in the mixed-clones pools.

关 键 词：云南药用野生稻 BIABC文库 混合克隆池 PCR筛选 XA21基因 

分 类 号：S511.9[农业科学—作物学] S567.219