To obtain the entire coding sequence of human PRL-3 gene and construct its prokaryotic expression vector. With total RNA extracted from the human colorectal carcinoma cell line SW480 as the template, PRL-3 gene was amplified by RT-PCR with primers designed according to the published sequence of GenBank, and the product was inserted into pGEM-T Easy vector. The recombinant plasmid pGEM-T-PRL-3 was identified by restriction endonuclease analysis and DNA sequencing. After digestion with restriction endonuclease, PRL-3 gene was cloned into the multicloning sites of the prokaryotic expression vector pGEX-4T-1. The entire coding region of human PRL-3 gene was cloned, and the recombinant pGEX-4T-1-PRL-3 vector was successfully constructed and expressed, which may provide the basis for further study of the relationship between human colorectal carcinoma and PRL-3 gene.