Team:Osaka/Tests

Test

Cell survival assay

As an initial screen for DNA repair protein activity , we used the UV assay. The cells were plated on respective agar plates at different dilutions, air dried, and then exposed to different doses of UV radiation. Plates were wrapped with aluminum foil and incubated in the dark. Colonyforming units were scored after 16h incubation at 37℃.

放射線耐性菌D.radioduranceのもつDNA-repair proteinであるPprI,PprA,PprM,RecAをそれぞれ導入した大腸菌にUV照射

Results

生存率のグラフ

PprM,RecAは生存率アップ、 I,Aは微妙な結果

PprIはRecAとPprAを誘導 PprAはRecAに依存しない修復機構をもっているらしい（変異が入った？） PprMは不明

RecA欠損株であるDH5αを用いたためPpr系の生存率は変化しなかったと思われる。 RecAを持つ株なら生存率上がってたかも。

Future work

We created some parts (PprI , PprA , PprM , RecA) but did not have time to evaluate them. Also, devices containing 2 or more DNA repair gene should have been constructed and assayed.

Reference

放射線抵抗性細菌の新規DNA修復促進タンパク質 , 佐藤勝也　その他 (2006)
PprA: a protein implicated in radioresistance of Deinococcus radiodurans stimulates catalase activity in Escherichia coli, Swathi Kota et al (2006)