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Response of bacteriophage T7 biological dosimeter to dehydration and extraterrestrial solar UV radiation

机译:T7噬菌体生物剂量计对脱水和地外太阳紫外线辐射的响应

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The experiment "Phage and uracil response" (PUR) will be accommodated in the EXPOSE facility of the ISS. Bacteriophage T7/isolated T7 DNA will be exposed to different subsets of extreme environmental parameters in space, in order to study the Responses of Organisms to the Space Environment (ROSE). Launch into orbit is preceded by EXPOSE Experiment Verification Tests (EVT) to optimize the methods and the evaluation. Bacteriophage T7/isolated T7 DNA thin layers were exposed to vacuum (10~(-6) Pa), to monochromatic (254 nm) and polychromatic (200-400 nm) UV radiation in air as well as in simulated space vacuum. Using neutral density (ND) filters dose-effect curves were performed in order to define the maximum doses tolerated. The effect of temperature fluctuation in vacuum was also studied. The structural/chemical effects on bacteriophage T7/isolated T7 DNA were analyzed by spectroscopic and microscopical methods. Characteristic changes in the absorption spectrum and in the electrophoretic pattern of phage/DNA have been detected indicating the damage of isolated and intraphage DNA. DNA damage was also determined by quantitative PCR (QPCR) using 555 and 3826 bp fragments of T7 DNA. We obtained substantial evidence that DNA lesions (e.g. strand breaks, DNA-protein cross-links, cyclobutane pirimidine dimers (CPDs) etc.) accumulate throughout exposure. Preliminary results suggest a synergistic action of space vacuum and UV radiation with DNA being the critical target.
机译:实验“噬菌体和尿嘧啶反应”(PUR)将容纳在国际空间站的EXPOSE设施中。噬菌体T7 /分离的T7 DNA将暴露于太空中极端环境参数的不同子集,以研究生物对太空环境(ROSE)的反应。在进入轨道之前,先进行EXPOSE实验验证测试(EVT),以优化方法和评估。将噬菌体T7 /分离的T7 DNA薄层暴露于空气中以及在模拟的空间真空中暴露于真空(10〜(-6)Pa),单色(254 nm)和多色(200-400 nm)UV辐射中。使用中性密度(ND)过滤器进行剂量-效果曲线,以定义可耐受的最大剂量。还研究了真空中温度波动的影响。通过光谱和显微镜方法分析了对噬菌体T7 /分离的T7 DNA的结构/化学作用。已经检测到噬菌体/ DNA的吸收光谱和电泳图谱的特征变化,表明分离的噬菌体DNA和噬菌体内DNA的破坏。还使用555和3826 bp的T7 DNA片段通过定量PCR(QPCR)确定了DNA损伤。我们获得了大量证据,证明DNA损伤(例如链断裂,DNA-蛋白质交联,环丁烷嘧啶二聚体(CPD)等)在整个暴露过程中积累。初步结果表明,空间真空和紫外线辐射具有协同作用,而DNA是关键目标。

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