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Team:Osaka/Tests
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== Test == | == Test == | ||
=== Cell survival assay === | === Cell survival assay === | ||
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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℃. | 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℃. | ||
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=== SOS promoter assay === | === SOS promoter assay === | ||
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Revision as of 05:28, 5 October 2011
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
SOS promoter assay
Results
Cell viability
グラフ
recA gene could induce high cell viability. RecA protein has key role of SOS response.
This result revealed that the cell inserted recA gene can get tolerance against DNA damage.
pprM gene could also bring high tolerance to inserted cells. discussion 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？ 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)
