目的　简化重组腺病毒载体的构造方法 ,构建含Smad3DcDNA或Smad7cDNA的重组腺病毒载体 ,为下一步的基因治疗奠定基础。方法　以AdEasySystem为基础 ,应用序贯化学转化方法 ,在大肠杆菌E .coliBJ5 183体内将携带目的基因的穿梭质粒和骨架质粒重组。结果　成功构建了重组腺病毒载体pAd Smad3D和 pAd Smad7及空腺病毒载体 ,并经酶切鉴定证实。结论　用序贯化学转化方法 ,可以快速高效地在大肠杆菌内构建重组腺病毒载体。 Objective To simplify the construction of recombinant adenovirus vector and construct a recombinant adenovirus vector containing Smad3DcDNA or Smad7 cDNA to lay the foundation for the next gene therapy. Methods Based on AdEasySystem, the shuttle plasmid and backbone plasmid carrying the target gene were recombined in E. coli BJ5 183 by sequential chemical transformation. Results The recombinant adenovirus vector pAd Smad3D, pAd Smad7 and empty adenovirus vector were successfully constructed and confirmed by restriction enzyme digestion. Conclusion Sequential chemical transformation method can construct recombinant adenovirus vector rapidly and efficiently in E. coli.