Cotton leaf curl virus (CLCuV) belongs to the subgroup III of geminiviruses with single strand DNA genome. Study demonstrated that the bidirectional promoter of CLCuV had activity in Agrobacterium tumefaciens (Smith et Townsend) Conn. This is the first report for the activity of the bidirectional promoter of geminivirus in A. tumefaciens. Results showed that the activity of the complementary sense promoter was stronger than that of virion sense promoter, and was detected 2-fold higher than that of CaMV 35S promoter in A. tumefaciens. Moreover, the promoter 5' deletion analysis indicated that the mean GUS activity driven by a 287 nucleotides complementary sense promoter fragment (from-287 to the translation initiation site) is 4 times higher than that driven by the whole complementary sense promoter in A. tumefaciens. This result suggested that there might exist negative regulatory elements in this deleted fragment. The function of other cis-elements included in CLCuV complementary sense promoter was also discussed in this paper.