Preliminary Study Of Effect On Radiation Resistance Of Yeast And Interaction Of Protein PprM, PprI And PprA From Deinococcus Radiodurans.

Background and purpose: Deinococcus radiodurans is known for its extreme radioresistance, it is a model organism to study DNA damage and repair.Ppr M is a new unique regulatory protein for radioresistance from Deinococcus radiodurans. In this study, we analyze effect on radiation resistance of yeast and interaction of the special protein Ppr M,Ppr I and Ppr A from Deinococcus radiodurans,so that we can probe the significant molecular mechanism that Ppr M which plays in radiation resistance of Deinococcus radiodurans and can study the role of radiation related proteins from Deinococcus radiodurans which play in eukaryotic.Methods: 1.We construct p GBKT7-ppr M, p GADT7-ppr I and p GADT7-ppr A vector, transform the three vectors into yeast Y187 respectively,transform p GBKT7-ppr M together with p GADT7-ppr I, p GBKT7-ppr M together with p GADT7-ppr A into yeast Y187 respectively, calculate survival rate of yeast strain Y187 compared with wild yeast,and make statistical analysis after different dose γ radiation.2.Transform p GBKT7-ppr M together with p GADT7-ppr I, p GBKT7-ppr M together with p GADT7-ppr A into yeast Y187 respectively, to verify whether there is interaction between Ppr M and Ppr I or Ppr A by yeast two-hybrid.Results: 1.We construct p GBKT7-ppr M,p GADT7-ppr I and p GADT7-ppr A vector successfully.When non-irradiated, Y187 yeast strains expressing Ppr M, Ppr I, Ppr A have no significant statistically difference in survival rate compared with wild yeast; at 50-200 Gy γ radiation, Y187 yeast strains expressing Ppr M, Ppr I, Ppr A, have significant statistically difference in survival rate compared with wild yeast;at 100-200 Gy γ radiation, expressing Ppr I Y187 yeast strain have significant statistically difference in survival rate compared with Ppr M-Ppr I co-expression in Y187 yeast strain; at 100-200 Gy γ radiation,expressing Ppr A Y187 yeast strain have significant statistically difference in survival rate compared with Ppr M-Ppr A co-expression in Y187 yeast strain. 2.After yeast two-hybrid verification we find that there is interaction between Ppr M and Ppr I or Ppr A.Conclusions: 1. Protein Ppr M can improve yeast radiation resistance, and in a higher dose γ radiation, it has synergistic effect with Ppr I and Ppr A. 2. There is interaction between Ppr M and Ppr I or Ppr A.