【摘要】 将盐生杜氏藻(6-4)光裂合酶基因Ds64PHR的编码序列构建到表达载体pET32a中,利用高效转化法将构建好的表达载体转入E.coli.CPD光裂合酶缺陷菌株SY2中,诱导表达(6-4)光裂合酶融合蛋白,对其功能进行验证.在含有Amp的LB平板上涂布相同数量的大肠杆菌,改变紫外照射强度、光照修复时间,统计最终的菌株存活率.经研究发现,在基因水平上,SY2中表达的盐生杜氏藻(6-4)光裂合酶具有修复紫外诱导损伤的功能,光照是修复功能实现的必需条件.更多还原

【Abstract】 Expression plasmid,pET32a-Ds64PHR,which was constructed by inserting(6-4) photoproducts photolyase gene from Dunaliella salina into E.coli.expression vector pET32a,was transformed into CPD-deleted(E.coli.SY2.)Then the gene was induced to express proteins and the function of the photolyase gene Ds64PHR was identified.The same amount of E.coli was pushed on the LB-Amp plates and the UV dose or the visible-light-irradiated time was chosen as a variable.Analysis of the number of bacteria suggest that the(6-4) PD photolyase expressed in SY2 had a function of repairing UV-induced lesions at gene level and light was necessary to this course.更多还原