李南羿; 姚占芳; 陳明杰
河南農業大學微生物教研室!鄭州450002; 河南農業大學微生物教研室!鄭州450002; 農業部食用菌遺傳育種重點開放實驗室!上海市農業遺傳育種重點開放實驗室; 上海市農業科學院食用菌研究所; 上海201106
【中文摘要】 采用CTAB/NaCl法提取草菇基因組DNA ,通過限制性內切酶Sau 3AI對純化的草菇基因組DNA進行部分酶切 ,獲得大小為 2 0kb左右的DNA片段。這些DNA片段再次純化后 ,與經EcoRI和BamHI雙酶切消化的LambdaGEM 11噬菌體DNA載體臂相連 ,以大腸桿菌(E .coli)LE392為寄主細胞 ,構建了草菇 (Volvariellavolvacea)基因文庫 ,所構建基因文庫效價為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.
【中文關鍵詞】 草菇; 噬菌體; 基因文庫
【英文關鍵詞】 Volvariella volvacea; Phage; Genomic library
【基金】上海市農委科技攻關重點項目資助項目編號:農科攻字 (98)第 0 1- 14號
【文獻出處】 食用菌學報,Acta Edulis Fungi,編輯部郵箱,2001年02期 【DOI】CNKI:SUN:SYJB.0.2001-02-002