Maize (Zea mays L.) is an important food crop and cultivates widely in the world. The Brittle 2 (Bt2) gene encodes the small subunit of ADP-glucose pyrophosphorylase (AGPase) enzyme, one of the enzymes that plays an important role in endosperm starch biosynthesis of cereal crops. Many studies around the world have demonstrated the successful transfer of the genes (Brittle 2 and Shrunken 2) encoding AGPase enzyme into plant to obtain the crop plants having enhanced starch biosynthesis. Bt2 gene has been transferred into maize plants and the content of starch in the transgenic plants was higher than that of wild-type, consequently, the yield of transgenic maize was increased significantly. To develop maize lines having high starch content by transgenic techniques, the construction of transformation vector carrying the Bt2 gene that involves in starch biosynthesis is an important prerequisite. The aim of this study is to construct the transformation vector carrying Bt2 for further gene transfer experiments. In this article, the authors present the results on construction of two transformation vectors pB1121/35S-Bt2-Nos and pCambia2300/Ubi-Bt2-Nos carrying Bt2 gene isolated from a maize line H20 and drived by 35S or Ubiquitin promoter. The complete constructs pB1121/35S-Bt2-Nos and pCambia2300/Ubi-Bt2-Nos were, sussessfully, transformed into E. coli strain DH5a and into A. tumefaciens strains C58 and EHA105. The presence of the target gene (Bt2) and its promoter (Ubi or 358) in E. coli strain DH5a and A. tumefaciens strains C58 and EHA105 were confirmed by colony-PCR amplification with respective specific primers and cleaving by suitable restriction enzymes. The constructed vectors will be further used in transfer of Bt2 gene into embryos or meristem for the regeneration of transgenic maize lines with enhanced starch biosynthesis and good quality.