Langeveld S A, Yasui A, Eker A P
Mol Gen Genet. 1985;199(3):396-400. doi: 10.1007/BF00330748.
A 2 kb DNA fragment, containing the photoreactivation gene phr1 from Escherichia coli, was inserted at the BamH1 site in the tet gene of the yeast--E. coli shuttle vector pJDB207. Photoreactivation--deficient Saccharomyces cerevisiae cells transformed with this plasmid showed photoreactivation of killing after UV irradiation of the cells, while extracts of transformed cells exhibited photoreactivating activity in vitro. Far more photoreactivating enzyme molecules were found when the gene was inserted in the plasmid in the opposite orientation to the tet gene as compared with a plasmid carrying the inserted gene in the same orientation. Photoreactivating enzyme encoded by the E. coli phr1 gene and produced in transformed yeast cells has characteristics of the E. coli photoreactivating enzyme (flavoprotein) as judged from the influence of ionic strength on photoreactivating activity.
一个含有来自大肠杆菌光复活基因phr1的2 kb DNA片段,被插入到酵母-大肠杆菌穿梭载体pJDB207的tet基因的BamH1位点。用该质粒转化的光复活缺陷型酿酒酵母细胞在紫外线照射后显示出杀伤的光复活,而转化细胞的提取物在体外表现出光复活活性。与携带插入基因同向的质粒相比,当该基因以与tet基因相反的方向插入质粒时,发现了更多的光复活酶分子。从离子强度对光复活活性的影响判断,由大肠杆菌phr1基因编码并在转化酵母细胞中产生的光复活酶具有大肠杆菌光复活酶(黄素蛋白)的特性。