As our previous study shown, the therapeutic effect of double genes (SERCA2a and Cx43) on heart failure after myocardial infarction (MI) were better than that of single gene (SERCA2a or Cx43) therapy on the basis of bone marrow stem cells (BMSCs) transplantation. Based on previous research, the aim of this study was to investigate the optimize ratio of co-delivery of SERCA2a and Cx43 genes for MI therapy after biotinylated microbubbles (BMBs) transplantation via ultrasonic-targeted microbubble destruction (UTMD). Forty rats underwent left anterior descending (LAD) ligation and BMSCs injection into the infarct and border zones. Four weeks later, double genes of SERCA2a and Cx43 with different ratios (1:1, 1:2 and 2:1) were co-delivered into the infarcted heart via the UTMD. Cardiac mechano-electrical function was determined at 4-wk after gene delivery, and the hearts of the rats were harvested for measurement of MI size and detection of SERCA2a and Cx43 expression. Q-PCR analysis of the expression of Nkx2.5 and GATA4 in the myocardial infarct zone and measurement of neovascularization in infarcted heart. After comparing the therapeutic effects of different co-gene ratios, the SERCA2a/Cx43-1:2 group shown remarkable cardiac electrical stability and strengthen the role of anti-arrhythmia. In conclusion, the optimum ratio of SERCA2a/Cx43 gene is 1:2, which take advantageous in maintaining cardiac electrophysiological stability.