Abstract:

Objective     To identify the promoter region of human tumor necrosis factor-α induced protein 8-like 2（TIPE2）, and important regulatory regions of a 1331bp putative TIPE2 promoter. Methods      A potential 1331bp promoter region and 7 fragments of 5′ truncation upstream TIPE2 gene were amplified from human genomic DNA by PCR. Then they were inserted into the luciferase report gene vector pGL4-Basic and transfected into human SKOV3 cells. Relative luciferase activity was detected to determine promoting activities. Results      DNA sequencing results indicated that promoter fragments were correctly identified. Analysis of relative luciferase activity showed that the 1331bp presented a strong promoter activity, which was increased by 7.9fold compared with pGL4-Basic.  TIPE2 promoter activity was respectively increased by 6.38-fold and 7.32-fold after -965bp to -593bp and -501bp to -390bp were deleted, while promoter activity was decreased by 2.04-fold after -593bp to -501bp was deleted. Conclusion     The 1331bp region upstream TIPE2 gene has high promoter activity. Two potential negative and one positive regulatory regions were identified by 5′truncation of  the 1331bp promoter region.

Key words: Genes, TIPE2；Promoter regions(genetics)； Luciferase； Gene expression regulation