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Human - manganese super oxide dismutase - cdna, its expression in bacteria and a method for the preparation of enzymatically active human - manganese super oxide dismutase

机译:人锰超氧化物歧化酶cdna及其在细菌中的表达和酶促活性人锰超氧化物歧化酶的制备方法。

摘要

A double-stranded cDNA molecule which includes DNA encoding human manganese super oxide 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 mole- cules may be introduced in procaryotic, e.g., bacteri- al, or eukaryotic, e.g. , yeast or mammalian, cells and the resulting cells cultured or grown under suitable conditions sa as to produce human manganese superoxide dismutase or analogs thereof which may then be recov- ered. Human MnSOD or analogs thereof may be used to catalyze the reduction of superoxide radicals, reduce reperfusion injury, prolong the survival time of iso- lated organs, or treat inflammations. The invention also concerns a method of producing enzy- matically active human manganese superoxide dismutase or an analog thereof in a bacterial cell which contains and is capable of expressing a DNA sequence encoding the superoxide dismutase by maintaining the bacterial cell under suitable conditions and in a suitable pro- duction 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. This invention also concerns a method of recovering purified enzymatically active manganese super oxide dismutase from bacterial cells.
机译:已经创建了包括编码人锰超氧化物歧化酶的DNA的双链cDNA分子。已经发现了编码人锰超氧化物歧化酶的双链DNA分子的一条链的序列。这些分子可以以原核的,例如细菌的或真核的,例如细菌的形式引入。酵母或哺乳动物的细胞,以及在合适条件下培养或生长的所得细胞,可产生人类锰超氧化物歧化酶或其类似物,然后再回收。人MnSOD或其类似物可用于催化超氧化物自由基的还原,减少再灌注损伤,延长离体器官的存活时间或治疗炎症。本发明还涉及在细菌细胞中产生酶活性的人锰超氧化物歧化酶或其类似物的方法,该细菌细胞包含并能够通过将细菌细胞保持在合适的条件和合适的条件下表达编码超氧化物歧化酶的DNA序列。生产介质。向生产培养基中添加一定量的Mn ++,以使培养基中Mn ++的浓度大于约2 ppm。本发明还涉及从细菌细胞中回收纯化的具有酶活性的锰超氧化物歧化酶的方法。

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