Fig. 5

Mini-chromosome loss assay. Transformants (constructed by splitting left and right edge of CgLEU2 replaced Chr2-6 region) harboring mini-chromosome consisting of only CgLEU2 and URA3 markers were cultured in liquid YPDA medium and subjected to dispense on YPDA plate after appropriate dilution. After colony formation, this plate was used as a master plate to replica plating on Ura minus, Leu minus, 5-FOA and YPDA media (as a control). After replica plating, two types of colonies were appeared in YPDA master plate, we named them as Type 1 and Type 2. To distinguish them, three Type 1 colonies were circled by red color and two Type 2 colonies were circled by green color. In other replica-plated plates, colonies that showed regular growth were circled by blue color and colonies that showed no growth were circled by yellow color. Type 1 colonies could not grow on Ura minus, Leu minus media but could grow on 5-FOA and YPDA media. On the other hand, Type 2 colonies could grow on Ura minus, Leu minus and YPDA media but could not grow on 5-FOA medium