Gruber M Y, Glick B R, Thompson J E
Department of Biology, University of Waterloo, ON, Canada.
Proc Natl Acad Sci U S A. 1990 Apr;87(7):2608-12. doi: 10.1073/pnas.87.7.2608.
The Mn superoxide dismutase gene of Escherichia coli was subcloned into the E. coli-Anacystis nidulans shuttle vector pSG111 to make the plasmid pMYG1. Transformation of E. coli HB101 with pMYG1 resulted in a 6-fold increase in superoxide dismutase activity. There was also induction of Mn superoxide dismutase in the transformants upon exposure to paraquat, as evidenced by dramatically increased levels of the Mn superoxide dismutase polypeptide in cytoplasmic extracts and a 16-fold further increase in superoxide dismutase activity. As well, the E. coli transformants showed resistance to paraquat-mediated inhibition of growth. Anacystis nidulans, a cyanobacterium that has no detectable Mn superoxide dismutase and is, consequently, very sensitive to oxidative stress, was also transformed with pMYG1. The transformants had detectable levels of Mn superoxide dismutase protein and showed resistance to paraquat-mediated inhibition of growth and photobleaching of pigments. Paraquat is known to promote formation of the superoxide radical anion, O2-., and thus the data have been interpreted as indicating that the cloned Mn superoxide dismutase provides protection in both E. coli and A. nidulans against damage attributable to O2-..
将大肠杆菌的锰超氧化物歧化酶基因亚克隆到大肠杆菌-集胞藻穿梭载体pSG111中,构建成质粒pMYG1。用pMYG1转化大肠杆菌HB101,超氧化物歧化酶活性提高了6倍。当用百草枯处理这些转化体时,锰超氧化物歧化酶也被诱导,这可从细胞质提取物中锰超氧化物歧化酶多肽水平的显著增加以及超氧化物歧化酶活性进一步提高16倍得到证明。此外,大肠杆菌转化体对百草枯介导的生长抑制具有抗性。集胞藻是一种蓝细菌,检测不到锰超氧化物歧化酶,因此对氧化应激非常敏感,它也用pMYG1进行了转化。转化体中可检测到锰超氧化物歧化酶蛋白水平,并且对百草枯介导的生长抑制和色素光漂白具有抗性。已知百草枯可促进超氧阴离子自由基O2-的形成,因此这些数据被解释为表明克隆的锰超氧化物歧化酶在大肠杆菌和集胞藻中均能提供保护,抵御由O2-造成的损伤。