吕作舟; 马爱民; 杨新美
华中农业大学; 上海农科院食用菌研究所; 华中农业大学 武汉 430070; 武汉 430070
【中文摘要】 本文采用原生质体裂解法制备高分子量平菇总DNA。通过Sau3A部分酶切和蔗糖密度梯度离心,获得15～25Kb平菇总DNA酶切片段。并以Lambda的衍生物EMBL3为载体,酶连形成重组DNA。然后将其包装,转染受体菌LE392,在TB平板上获得2.5×104个重纽子克隆。此重组子克隆群即构成中蔬10号平菇菌株基因组文库。
【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.
【中文关键词】 平菇; 原生质体裂解法; 高分子量总DNA; EMBL3载体; 基因组文库
【英文关键词】 fleurotus ostreatus Protoplastio lysis method High-molecular-weight total DNA EMBL3 vector Genomic library
【基金】国家自然科学基金
【文献出处】 中国食用菌,Edible Fungi of China,编辑部邮箱,1992年06期 【DOI】CNKI:SUN:ZSYJ.0.1992-06-007