Objective:To construct lentiviral expression vector contained phosphatidylinositol 3-kinase p110 gamma(PI3KCG), and identify its expression in neonatal rat cardiomyocytes and detect its prelimary function. Methods:The PI3KCG lentiviral vector plasmid (PLV-PI3KCG)was constructed by homologous recombination method. The three plasmids of PLV-PI3KC and of lentivirus system were co-transfected into human embryonic kidney 293T cells by using lipofectamine. The expression of green fluorescent protein (GFP)in the control group was examined by using fluorescent microscope at 24 h and 48 h after transfection. Recombinant PLV-PI3KCG plasmid was successfully identified in 293T cells detected by polymerase chain reaction (PCR). The viral supernatant was collected with 72 h after transfection. The ischemia/reoxygenation (I/R)injury model of neonatal rat myocardial cells was established. Myocardial cells isolated from SD neonatal rats were random division into five groups after being cultured for 3d: the normal control group, the I/R group, the null vector positive control group, the PI3KCG transfection preconditioning group and the PI3KCG transfection+ Ly294002 group. Various techniques were adopted to detect the products of cells and cellular:the cardiomycytes beat frequency, the levels of myocardial cells viability rate and the levels of lactate dehydrogenase (LDH). Results:The PLV-PI3KCG plasmid was constructed and expressed in the 293T cells successfully. Compared with the I/R group, the myocardial cells viability and cardiomycytes beat frequency of the PI3KCG transfection preconditioning group were significantly increased (P < 0.01,respectively), and released LDH were significantly decreased (P < 0.01,respectively). Conclusion:PLV-PI3KCG plasmid was expected to become an available vector to investigate PI3K/Akt pathway in the cardiocytes ischemia reperfusion injury process.