Abstract: Objective To construct and express the recombinant plasmid of DLC-1(deleted in liver cancer). Methods DLC-1 gene fragment was obtained by PCR. DLC-1DNA fragment was digested with Xba Ⅰ and BamH Ⅰ together, and then ligation reaction to eukaryotic expression vector pcDNA3.1 which was also digested under the same condition. The product of ligation reaction was transformed into Escherichia coli DH5a. Recombinant plasmid pcDNA3.1/DLC-1 was transfected into HT-29 cells with liposome. Then, the product of recombinant plasmid was obtained by RT-PCR. Results Double restriction enzyme digestion and subsequent sequencing of recombinant plasmids confirmed the correctness of the recombinant plasmids. Recombinant plasmid pcDNA3.1/DLC-1 was transfected into HT-29 cells with liposome, and the product of recombinant plasmid was obtained. Conclusion The pcDNA3.1/DLC-1 was expressed in HT-29. This method provides a basis for studying colorectal cancer.