Aim To construct recombinant adeno-associated viruses (rAAV) vectors carrying human apolipoprotein A_Ⅰ (apoA_Ⅰ) and apo A_Ⅰ Milano cDNA, and explore a new strategy to prevent and treat the atherosclerotic diseases. Methods Human apo A_Ⅰ cDNA with a his-tag in upward of the cDNA sequence was obtained by reverse transcription-polymerase chain reaction (RT-PCR) and polymerase chain reaction (PCR), human apo A_Ⅰ Milano cDNA was then prepared by Site-Directed Mutagenesis. The particle numbers of rAAV vectors after extractd with a most economic and convenient method was assayed by Dot-blot, and the purity was assayed by SDS polyacrylamide gel electrophoresis (SDS-PAGE). The expression efficiency of the apo A_Ⅰ and apo A_Ⅰ Milano in C₂C₁₂ after infected by rAAV vectors were detected by ELISA method. Results The titre of the rAAV vectors of apo A_Ⅰ and apo A_Ⅰ Milano was about 2×10 14/L, and the result of SDS-PAGE showed the purity of the rAAV vectors was good. The expressed apo A_Ⅰ level is 0.39±0.04 mg/L and the apo A_Ⅰ Milano is 0.31±0.03 mg/L in the DMEM culture medium. Conclusions The success of the rAAV vectors construction and purification and the expression of apo A_Ⅰ and apo A_Ⅰ Milano in C2C 12 cells mediated by these vectors, makes the injection of rAAV encoding human apo A_Ⅰ (rAAVA) and rAAV encoding human apo A_Ⅰ milano (rAAVAM) vectors in mouse muscular cells possible, and contributes to the hope of finding a safe and effective way to prevent and treat atherosclerotic diseases.