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| 云南药用野生稻BIBAC文库混合克隆池制备及筛选 | |
| 引用本文： | 刘艳平,程在全,殷富有,余腾琼,张敦宇,郭怡卿,张薇,黄兴奇. 云南药用野生稻BIBAC文库混合克隆池制备及筛选[J]. 生物技术通报, 2012, 0(2): 117-120 |
| 作者姓名： | 刘艳平 程在全 殷富有 余腾琼 张敦宇 郭怡卿 张薇 黄兴奇 |
| 作者单位： | 1. 云南省农业科学院生物技术与种质资源研究所,昆明650223;云南大学生命科学学院,昆明650091 2. 云南省农业科学院生物技术与种质资源研究所,昆明,650223 3. 云南省昭通市农科所,昭通,657000 |
| 基金项目： | 国家自然科学基金,国家公益性行业(农业)科研专项,云南省应用基础研究重点项目,云南省高端人才引进项目,云南省攻关项目 |
| 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. |
| 关 键 词： | 云南药用野生稻 BIABC文库 混合克隆池 PCR筛选 Xa21基因 |
Preparation and Screening of Mixed-clones Pools from BIBAC Library of Oryza officinallis in Yunnan Province |
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| Liu Yanping , Cheng Zaiquan , Yin Fuyou , Yu Tengqiong , Zhang Dunyu , Guo Yiqing , Zhang Wei , Huang Xingqi. Preparation and Screening of Mixed-clones Pools from BIBAC Library of Oryza officinallis in Yunnan Province[J]. Biotechnology Bulletin, 2012, 0(2): 117-120 | |
| Authors: | Liu Yanping Cheng Zaiquan Yin Fuyou Yu Tengqiong Zhang Dunyu Guo Yiqing Zhang Wei Huang Xingqi |
| Affiliation: | 1,2(1Biotechnology & Genetic Germplasm Institute,Yunnan Academy of Agricultural Sciences,Kunming 650223;2College of Life Science,Yunnan University,Kunming 650091;3Zhaotong Agricultural Sciences Institute,Zhaotong 657000) |
| Abstract: | 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. |
| Keywords: | Oryza officinallis Wall.ex Watt.in Yunnan BIBAC library Mixed-clones pools PCR screening Xa21 gene |
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