Maps of loop-in cdc48 ts alleles

These plasmids are cut with XhoI to make tandem copies of CDC48. This makes one containing mutations to a cdc48 ts allele and then also truncates the other copy so that the sole functional copy of CDC48 is the ts allele. When using, it is important to transform onto -Ura3 plates and grow at 25C for quite a while. The small colonies you get are the ts ones typically使用XhoI对质粒进行酶切，以制备CDC48的串联重复拷贝。经该酶切处理后，其中一个拷贝携带cdc48 ts（温度敏感）等位基因突变，另一个拷贝则被截短，使得CDC48唯一的功能性拷贝即为该ts等位基因。使用该质粒体系时，需将其转化至-Ura3筛选平板，并在25℃下培养较长时长。通常所获得的小型菌落即为携带ts等位基因的转化菌株。