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In addition to its antioxidant activity aimed at decreasing oxidative stress, O 2 -O 3 therapy could improve the metabolism of erythrocytes and O 2 supply to hypoxic tissues by improving glycolysis with the formation of 2,3-diphosphoglyceric acid (2,3-DPG) and ATP, increasing Krebs cycle and leading to the shift the HbO 2 dissociation curve to the right (Giunta et al., 2001)

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Select several individual colonies that are positive by PCR and grow a separate 5 ml mini-culture of each selected transformant to screen for expression of the target protein (see Note 6)