耐辐射球菌pprI基因转染对受照酵母菌抗氧化能力的影响  

Effect of antioxident capacity on irradiated Pichia pastoris by the transfection of Deinococcus radiodurans pprI gene

作　　者：丛瑛[1] 吴伟[1] 文玲[1] 杨占山[1] 

机构地区：[1]苏州大学医学部放射医学与防护学院放射毒理教研室 江苏省放射医学与防护重点实验室,苏州215123

出　　处：《辐射研究与辐射工艺学报》2013年第4期31-34,共4页Journal of Radiation Research and Radiation Processing

基　　金：国家自然科学基金项目(81072236);国防基础科研资助项目(A3820060138);江苏省高校优势学科建设工程项目(PAPD)资助

摘　　要：研究不同剂量60Coγ射线对转染了耐辐射球菌pprI基因的毕赤酵母菌过氧化氢酶(CAT)、超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)活性的影响,探讨耐辐射球菌pprI基因在酵母菌中的表达对辐射细胞抗氧化能力的作用。Western blot检测酵母菌培养上清液中目的蛋白PprI的表达分泌。分别通过可见光法、单胺氧化法、分光光度比色法测定CAT、SOD和GSH-Px的活性。转染耐辐射球菌pprI基因的毕赤酵母菌中检测到PprI蛋白质的表达。当γ射线剂量为2Gy、4Gy、6Gy、8Gy、10Gy时,不论是转基因毕赤酵母组还是对照组,CAT、SOD、GSH-Px活性均随着剂量的增加出现先升高后降低的趋势,并且受照后转基因毕赤酵母菌中3种酶的活性均显著高于对照组(p<0.01)。耐辐射球菌pprI基因在酵母菌中的表达能显著增强辐射细胞的抗氧化能力,这可能是原核pprI基因转染增强真核细胞抗辐射能力的机理之一。The activity effect of catalase （CAT）, the superoxide dismutase （SOD） and the glutathione peroxidase （GSH-Px） of Pichia pastoris transfected by Deinococcus radiodurans pprI gene in different doses of^60Coγ-ray are studies. The role of Deinococcus radiodurans pprI gene expression in Pichia pastoris on the antioxidant ability of irradiated cells. The target protein secretion in the culture supernatant of Pichia pastoris was verified by Western blot. The activity of catalase （CAT）, superoxide dismutase （SOD） and glutathione peroxidase （GSH-Px） was measured by visible absorption spectrometry method, monoamine oxidase method and spectrophotometry colorimetric method, respectively. The expression of Pprl protein is detected in the Deinococcus radiodurans pprI gene transfected Pichia pastoris. The results show that the activities of CAT, SOD and GSH-Px all increase firstly and then decrease with the increasing absorbed doses at range of2Gy, 4Gy, 6Gy, 8Gy, 10Gy in both transgenic Pichia pastoris and control groups . And after irradiation, the activities of these three enzymes in transgenic Pichia pastoris are significantly higher than that in the control group （p〈0.01）. The expression of Deinococcus radiodurans pprI gene in Pichia pastoris can obviously increase the antioxidant ability of irradiated cells, which may be one of the mechanisms that prokaryotic pprl gene transfection enhances the radioresistance ability of eukaryotic cells.

关 键 词：耐辐射球菌pprI 毕赤酵母 60Coγ射线 过氧化氢酶 超氧化物歧化酶 谷胱甘肽过氧化物酶 

分 类 号：TL71[核科学技术—辐射防护及环境保护]