| abstract |
A double-stranded cDNA molecule which includes DNA encoding human manganese superoxide dismutase has been created. The sequence of one strand of a double-stranded DNA molecule which encodes human manganese superoxide dismutase has been discovered. Such molecules may be introduced in prokaryotic, e.g., bacterial, or eucaryotic, e.g., yeast or mammalian, cells and the resulting cells cultured or grown under suitable conditions so as to produce human manganese superoxide dismutase, human manganese superoxide dismutase analogs or human manganese superoxide dismutase mutants which may then be recovered. By this invention, human manganese superoxide dismutase gene fragments from various plasmids may be ligated to yield a complete genomic human manganese superoxide dismutase gene fragment. Human manganese superoxide dismutase, analogs, or mutants may be used to catalyze the reduction of superoxide radicals, reduce reperfusion injury, prolong the survival time of isolated organs, or treat inflammations. n The invention also concerns a method of producing enzymatically active human manganese superoxide dismutase compounds in a bacterial cell which contains and is capable of expressing a DNA sequence encoding the superoxide dismutase compound by maintaining the bac terial cell under suitable conditions and in a suitable production medium. The production medium is supplemented with an amount of Mn⁺⁺ so that the concentration of Mn⁺⁺ in the medium is greater than about 2 ppm. Genomic manganese superoxide dismutase DNA should also be capable of expression in eucaryotic cells under suitable conditions. n This invention also concerns a method of recovering purified enzymatically active manganese superoxide dismutase, analogs or mutants from bacterial cells. It should also be possible to recover manganese SOD from genomic manganese superoxide dismutase DNA expressed in eucaryotic cells using similar methods. n The invention further concerns uses of the human manganese superoxide dismutase, human manganese superoxide dismutase analogs or human manganese superoxide dismutase mutants, various new analogs, compositions of the compounds and methods of treating various disorders using these compositions. |