用λZAP Express作载体构建灵芝原基表达型cDNA文库  

Construction of cDNA expression library of Ganoderma lucidum primordium using λ ZAP Express vector

作　　者：赵明文[1] 李娜[1] 梁婉琪[2] 王南[3] 张大兵[2] 潘迎捷[3] 

机构地区：[1]南京农业大学农业部农业环境微生物工程重点开放实验室,江苏南京210095 [2]上海市农业科学院生物技术中心 [3]上海市农业科学院食用菌研究所

出　　处：《南京农业大学学报》2004年第4期64-68,共5页Journal of Nanjing Agricultural University

基　　金：国家自然科学基金 (3 0 3 0 0 0 0 6);南京农业大学青年基金 (KJ0 3 0 0 4);上海市科委重点专项经费

摘　　要：根据对韩国灵芝菌丝体、不同发育阶段子实体及孢子粉中灵芝酸含量的测定结果 ,以含量较高的灵芝原基为材料 ,提取mRNA ,从而构建以λZAPExpress为载体的表达型cDNA文库。经检测 ,所构建文库的滴度达到 6× 10 5pfu·mL-1,重组率达到 96 % ,插入片段的大小均在 70 0bp以上。该表达文库的构建 ,为进一步克隆和鉴定灵芝酸合成的相关基因打下了良好的基础。Firstly,ganoderic acid contents of Ganoderma lucidum HG at different developmental stages were detected.Thus,primordium with relatively high level of ganoderic acid contents was chosen as material to isolate mRNA for constructing cDNA expression library using λ ZAP Express vector.Further results showed that the tilter of the cDNA expression library was 6×105 pfu·mL -1,while the recombinant rate reached to 96%,and the length of inserts was all above 700 bp.Therefore,the constructed cDNA expression library is helpful in the further study on the cloning and characterization of the ganoderic acid synthesis related gene in G.lucidum.

关 键 词：灵芝酸 原基 CDNA表达文库 

分 类 号：Q785[生物学—分子生物学] S646.2[农业科学—蔬菜学]