稻瘟病菌细菌人工染色体(BAC)基因组文库的构建

The Construction of BacterialArtificial Chromosom (BAC) Library of Rice Blast Pathogen

稻瘟病菌是一种重要的植物病原菌 ,同时又是研究植物和病原物之间相互关系的一个重要试验系统。实验通过采用 95 - 2 3- 4a稻瘟病菌株经提取纯化得到该稻瘟病菌的细胞核基因组总DNA ,用限制性内切酶部分酶解后 ,与经末端去磷酸化处理过的pCUGIBAC1质粒载体按一定摩尔比例相互连接 ,通过转化E .coliDH10B感受态细胞进而通过蓝白斑筛选挑取白色克隆从而构建了 95 - 2 3- 4a稻瘟病菌株的细菌人工染色体 (BAC)基因组文库。通过进行酶切检测及Southern杂交分析后确定所构建的BAC文库平均插入片段 2 9.1kb ,该文库覆盖 7.4倍基因组 。

The fungus Magnaporthe grisea is an important plant pathogen and is a useful model organism for studying various aspects of host pathogen interactions. The fungus infects a wide range of grasses, including barley and millet, but it is best known as the causal agent of the blast disease of rice ( Oryza sativa ).The authors have constructed a genomic DNA library using the bacterial artificial chromosome(BAC) with Genomic DNA digested HindⅢ was ligated to the BAC vector ,and the ligation mixture was used to transform the bacterial strain DH10B.A total of over 11904 clones were obtained with the average insert size of about 29.1kb, 7.4×covering haploid genome of the fungus.The BAC library can be used for cloning of pathogenical gene.