李南羿; 姚占芳; 陈明杰
河南农业大学微生物教研室!郑州450002; 河南农业大学微生物教研室!郑州450002; 农业部食用菌遗传育种重点开放实验室!上海市农业遗传育种重点开放实验室; 上海市农业科学院食用菌研究所; 上海201106
【中文摘要】 采用CTAB/NaCl法提取草菇基因组DNA ,通过限制性内切酶Sau 3AI对纯化的草菇基因组DNA进行部分酶切 ,获得大小为 2 0kb左右的DNA片段。这些DNA片段再次纯化后 ,与经EcoRI和BamHI双酶切消化的LambdaGEM 11噬菌体DNA载体臂相连 ,以大肠杆菌(E .coli)LE392为寄主细胞 ,构建了草菇 (Volvariellavolvacea)基因文库 ,所构建基因文库效价为1.5× 10 5pfu/mL。
【英文摘要】 Genomic DNA of Volvariella volvacea was extracted by CTAB/NaCl method. While 20kb DNA fragments were obtained by partial digestion of purified genomic DNA of V.volvacea with restriction enzyme Sau 3AI. Repurified 20kb DNA fragments were ligated with Lambda GEM 11 Phage DNA vector arms digested by EcoR I and BamH I. And a genomic DNA library of V.volvacea was constructed with E.coli LE392 as host cell. The titer of the genomic DNA library was 1.5×10 5 pfu/mL.
【中文关键词】 草菇; 噬菌体; 基因文库
【英文关键词】 Volvariella volvacea; Phage; Genomic library
【基金】上海市农委科技攻关重点项目资助项目编号:农科攻字 (98)第 0 1- 14号
【文献出处】 食用菌学报,Acta Edulis Fungi,编辑部邮箱,2001年02期 【DOI】CNKI:SUN:SYJB.0.2001-02-002
