Ascenzioni F, Lipps H J
Gene. 1986;46(1):123-6. doi: 10.1016/0378-1119(86)90174-5.
A linear plasmid was constructed in vitro using the telomeres of the rDNA of Tetrahymena pyriformis. These telomeres were added to a yeast circular vector containing an ARS sequence from Dictyostelium, the LEU2 gene of yeast and the neo gene from Escherichia coli Tn5 fused with a eukaryotic promoter. The resulting plasmid was used to transform yeast. During the replication of the linear plasmid in yeast it was spontaneously modified at the extremity by the addition of 300 bp of yeast telomeric sequence for each end. Total DNA prepared from yeast transformants was used to transform the hypotrichous ciliate Stylonychia lemnae. The same plasmid isolated from Stylonychia can again be replicated in yeast.
利用梨形四膜虫rDNA的端粒在体外构建了一个线性质粒。将这些端粒添加到一个酵母环状载体中，该载体包含来自盘基网柄菌的ARS序列、酵母的LEU2基因以及与真核启动子融合的大肠杆菌Tn5的新霉素基因。所得质粒用于转化酵母。在酵母中线性质粒复制期间，其末端会自发修饰，两端各添加300 bp的酵母端粒序列。从酵母转化体中制备的总DNA用于转化下毛目纤毛虫莱氏全列虫。从莱氏全列虫中分离出的相同质粒可再次在酵母中复制。
