Molecular cloning and expression of two cytosolic copper-zinc superoxide dismutases genes from Nelumbo nucifera.

Two cytosolic copper-zinc superoxide dismutase (cytCuZnSOD) complementary deoxyribonucleic acid were achieved in Nelumbo nucifera (Elian). The active sites and common characteristics of cytCuZnSOD family were showed by homology modeling. The two recombinant proteins expressed by PET-32a vector showed the similar SOD activity (89.94 ± 0.54 U/mg) and could maintain more than 90% activity after incubation at 65°C. The subcellular location by green fluorescent protein revealed that these two isoforms were all located in cytosol and nucleus. The cytCuZnSODs were expressed in various parts of N. nucifera, which were expressed highest in the leafstalks and young leaves and lowest in the roots. The cytCuZnSOD messenger ribonucleic acids isolated from wounded leaves significantly increased at 1.5 h after treatment (HAT) with the highest expression at 3 HAT, after which the level decreased.