Grant-in-Aid for Young Scientists (B)
The yeast haploid strain(MATα) was transformed with wild-type p53 expressing plasmid using HIS and URA3 makers. On the other hand, the p53 missense mutation library was constructed by introducing the EGFP reporter plasmid and each of the 2314 mutant p53 expressing plasmids into the yeast haploid strain(MATa) using TRP1(for EGFP plasmids) and LEU2(for mutant p53 plasmids) makers in a comprehensive manner. The yeasts harboring mutant p53 expressing plasmid and EGFP reporter plasmid were mated with the wild-type p53 expressing yeast on the YPD plates. The mating diploid cells were transferred to the SC-trp-leu-his-ura plates and 5-FOA-containing SC-trp-leu-his plates. We investicated the colony formation of each diploid cells and screened the dominant-negative p53 mutants.
Attribution of KAKENHI