Nettleton C J, Bull C, Baldwin T O, Fee J A
Proc Natl Acad Sci U S A. 1984 Aug;81(15):4970-3. doi: 10.1073/pnas.81.15.4970.
A mixed-sequence synthetic oligodeoxynucleotide probe was used to identify clones within the Escherichia coli genomic library of Clarke and Carbon having an extrachromosomal copy of iron superoxide dismutase. Plasmids pLC13-47 and pLC18-11 were shown to contain the structural gene of the iron superoxide dismutase and to overproduce this protein under conditions of chloramphenicol amplification of plasmid copy number. The activities of both manganese and iron proteins were measured in extracts of host cells and plasmid-bearing cells grown over a wide range of oxygenation. The results confirm previous demonstrations that the manganese protein is repressed under anaerobic conditions and induced in the presence of oxygen. Induction of the manganese protein with increasing oxygenation was quantitatively similar in cells differing approximately equal to 7-fold in iron superoxide dismutase, suggesting that the intracellular concentration of superoxide might not be responsible for regulating synthesis of the manganese-containing superoxide dismutase.
使用混合序列合成寡脱氧核苷酸探针,从克拉克和卡尔文构建的大肠杆菌基因组文库中鉴定出携带铁超氧化物歧化酶染色体外拷贝的克隆。质粒pLC13 - 47和pLC18 - 11被证明含有铁超氧化物歧化酶的结构基因,并在氯霉素扩增质粒拷贝数的条件下过量产生这种蛋白质。在广泛的氧合条件下培养的宿主细胞和携带质粒的细胞提取物中,测定了锰蛋白和铁蛋白的活性。结果证实了先前的研究结果,即锰蛋白在厌氧条件下受到抑制,在有氧存在时被诱导。在铁超氧化物歧化酶含量相差约7倍的细胞中,随着氧合增加对锰蛋白的诱导在数量上相似,这表明超氧化物的细胞内浓度可能与含锰超氧化物歧化酶的合成调节无关。