Glumoff V, Käppeli O, Fiechter A, Reiser J
Institut für Biotechnologie, ETH-Hönggerberg, Zürich, Switzerland.
Gene. 1989 Dec 14;84(2):311-8. doi: 10.1016/0378-1119(89)90505-2.
An efficient transformation system for the filamentous yeast, Trichosporon cutaneum, has been developed. Transformation was obtained with plasmids carrying either the Escherichia coli hygromycin B phosphotransferase-encoding gene (hph) or the Streptoalloteichus hindustanus phleomycin-resistance gene (ble), as dominant selection markers. Expression of both resistance-conferring genes was controlled by the gpd promoter and the trpC terminator, from Aspergillus nidulans. The transformation frequency was up to 500 colonies/micrograms of transforming DNA, using the ble gene, and up to 100 colonies/micrograms of transforming DNA, using the hph gene. Co-transformation frequencies using unselected DNA varied between 50 and 65%. The transforming DNA was found to consist of multiple tandem plasmid copies of high Mr. This polymeric structure, in nonselective media, was mitotically unstable, possibly indicating that it existed in an episomal state.
已开发出一种用于丝状酵母皮状丝孢酵母的高效转化系统。使用携带大肠杆菌潮霉素B磷酸转移酶编码基因（hph）或印度链霉菌博来霉素抗性基因（ble）的质粒作为显性选择标记获得了转化。赋予抗性的两个基因的表达均由构巢曲霉的gpd启动子和trpC终止子控制。使用ble基因时，转化频率高达500个菌落/微克转化DNA，使用hph基因时，转化频率高达100个菌落/微克转化DNA。使用未选择的DNA的共转化频率在50%至65%之间。发现转化DNA由多个高分子量的串联质粒拷贝组成。这种聚合结构在非选择性培养基中是有丝分裂不稳定的，这可能表明它以附加体状态存在。
