Abstract: Using a system of multi-gene plant expression vector based on a set of Cre-LoxP recombination system, the BtCryIAc, BADH, pttGA20ox and rolB genes were inserted into the plant expression vector pYL1305, which was named pYL1305BBGR. The tabacco was transformed by leaf disc method mediated by Agrobacterium tumefaciens. Tested by PCR and Southern blot, the multi genes have been successfully transferred into the genome of tobacco. The semi-quantitative RT-PCR analysis showed that all the 4 genes were expressed normally.