Abstract:

Objective  To construct the human programmed cell death 4 promotor luciferase reporter gene vector and detect its activity in cells. Methods  The PDCD4 promotor from human genomic DNA was amplified by PCR, and was inserted into the luciferase report gene pGL4-basic vector. The amplified DNA sequence was confirmed by sequencing. To detect the transcriptional activity of human PDCD4-P1 in the plasmid,transient transfection was performed in different cell lines, and pRL-TK was used to determine the transfection efficiency. Results  The sequencing results indicated that the amplified sequence was correct. The results of transient transfection showed that the recombinant plasmid could be highly expressed in the OVCAR3 cell line which could highly express endogenous PDCD4, but lowly in the SKOV3 cell line in which lowly expressed endogenous PDCD4 could be detected. Conclusion  The human PDCD4 promotor luciferase reporter gene vector has been constructed successfully, which will lay experimental foundation for further study.

Key words: Programmed cell death 4; Promotor; Luciferase; Reporter gene