Construction of Eukaryotic Expression Vector of RNA interference Specific for Bcr/abl

Objective:To construct eukaryotic expression vector of siRNA specific for bcr/abl as a molecular target tool to cleave bcr/abl mRNA in chronic myelogenous leukemia cell, and to expore the feasibility of it. Methods:Genome sequences of bcr/abl fusion gene was retrieved from Genbank, siRNA(small interfering RNA)was designed according to the Tuschl's principle of RNAi-based medicine,and was converted into cDNA coding expression of shRNA(small hairpin RNAs)of siRNA for bcr/abl fusion gene.The cDNA was synthesized and inserted into plasmid pTER.The pTER shRNA1 and pTER shRNA2 of recombinant plasmid being eukaryotic expression vector was controlled by the H1 promoter of RNA polymerase Ⅲ,and identified by the restriction map and the sequence analysis.Results:The pTER/shRNA1 and pTER/shRNA2 of recombinant plasmid identificated by the restriction map and the sequence analysis completely coincided with the designs.Conclusions:The siRNA eukaryotic expression vector against bcr/abl mRNA has been successfully conctructed.