Aquaculture farming may benefit from genetically engineering fish to tolerate environmental stress. Here, we used the vector pCVCG expressing the Vitreoscilla hemoglobin (vhb) gene driven by the common carp beta-actin promoter to create stable transgenic zebrafish. The survival rate of the 7-day-old F-2 transgenic fish was significantly greater than that of the sibling controls under 2.5% O-2 (dissolved oxygen (DO), 0.91 mg/l). Meanwhile, we investigated the relative expression levels of several marker genes (hypoxia-inducible factor alpha 1, heat shock cognate 70-kDa protein, erythropoietin, beta and alpha globin genes, lactate dehydrogenase, catalase, superoxide dismutase, and glutathione peroxidase) of transgenic fish and siblings after hypoxia exposure for 156 h. The expression profiles of the vhb transgenic zebrafish revealed that VHb could partially alleviate the hypoxia stress response to improve the survival rate of the fish. These results suggest that that vhb gene may be an efficient candidate for genetically modifying hypoxia tolerance in fish.