Abstract: To construct a luciferase reporter plasmid containing human T-cell immunoglobulin- and mucin-domain-containing molecule-3(Tim-3) gene promoter． Methods The human Tim-3P2 was amplified by PCR using human genomic DNA as a template and was directionally cloned into pGL3basic multiple cloning sites to construct the luciferase reporter plasmid pGL3-Tim-3P2. To detect the transcriptional activity of human Tim-3P2 in the plasmid, transient transfection was performed in different cell lines. pRL-TK was used to normalize the transfection efficiency. Results DNA sequencing verified the successful construction of the plasmid pGL3-Tim-3P2. The results of transient transfection showed that the recombinant plasmid could be lowly expressed in the murine macrophage cell line RAW264.7 and murine melanoma cell-line-B16, both of which can endogenously express Tim-3， but not in the African green monkey kidney cell line COS-7 in which no endogenous Tim-3 could be detected. ConclusionThe recombinant plasmid containing human Tim3P2 has been successfully constructed which could selectively express in Tim-3 positive cells.

Key words: Genes, Tim-3, Human, Promoter, Luciferase reporter gene