In the study, a plant binary expression vector, named pCRI1210, was constructed on the basis of pBI121 vector, which contained CaMV 35S promoter, cotton β-tubulin gene introns , CaMV 35S polyA terminator and interference expression box with GFP as reporter gene. Functional analysis in tobacco showed that the interference express cassette can normally expressed exogenous gene and GFP gene . The vector had multiple cleavage sites in a TUB intron 5 "and 3" , so that target gene can be positive and reverse inserted and hairpin structure of dsRNA was builded. In addition, the vector carries multiple cloning sites at both 5′and 3′of the CaMV 35S promoter, which allows easy exchange 35S promoter to study other promoter functions. pCRI1210 can used to expression vector or interference vector,which provided a very practical tool for transgenic research.