呂作舟; 馬愛民; 楊新美
華中農業大學; 上海農科院食用菌研究所; 華中農業大學 武漢 430070; 武漢 430070
【中文摘要】 本文采用原生質體裂解法制備高分子量平菇總DNA。通過Sau3A部分酶切和蔗糖密度梯度離心,獲得15～25Kb平菇總DNA酶切片段。并以Lambda的衍生物EMBL3為載體,酶連形成重組DNA。然后將其包裝,轉染受體菌LE392,在TB平板上獲得2.5×104個重紐子克隆。此重組子克隆群即構成中蔬10號平菇菌株基因組文庫。
【英文摘要】 High-molecular-welght DNA of Pleurotus ostreatus was extracted using the method of protoplasic lysis. From this DNA, 15—25 kb fragments-were obtained by means of partial digestion with Sau 3A and ultraeentrifugation in saccharose gradient. The EMBL 3 of the Lambda derivation was used as the vector for ligating the 15—25 kb foreign DNA fragments and the recombinated DNA was formed. Then, the ligated DNA was packaged and transfected intoE, coli LE392. About 2.5×104 recombinanted clones, which contained the foreign DNA, were found on the TB medium plates. These clones are the genomic library of the Pleurotue ostreatus isolate Z S 10.
【中文關鍵詞】 平菇; 原生質體裂解法; 高分子量總DNA; EMBL3載體; 基因組文庫
【英文關鍵詞】 fleurotus ostreatus Protoplastio lysis method High-molecular-weight total DNA EMBL3 vector Genomic library
【基金】國家自然科學基金
【文獻出處】 中國食用菌,Edible Fungi of China,編輯部郵箱,1992年06期 【DOI】CNKI:SUN:ZSYJ.0.1992-06-007