Objective: To study the base sequence of an enhancer in up-stream 5'-flank near regulation region (from -595 to +74) of human colorectal cancer related gene ST13.
Methods: Several deletion PCR primers were designed. Amplified DNA fragments of ST13 gene 5'-flank near region were cloned with pGEMT-EASY vector and sequenced; then subcloned into several pGL2 report vectors respectively. Equal quantitative recombined DNA was transfected into SW620 cell lines and the luciferase activity was checked.
Results: Several amplified base sequence fragments (669 bp,263 bp,163 bp) in pGL2-Basic all enhanced and promoted luciferase gene expression strongly. The 47 bp and 101 bp fragments didn't promote luciferase gene expression. 101 bp fragment recombined with pGL2-Promoter enhanced luciferase gene expression distinctly (P<0.01), but the effect was less strong than the positive pGL2-Control(P<0.05).
Conclusion: The base sequence 101 bp (from -595 to -494) in up-stream 5'-flank near regulation region of colorectal cancer related gene ST13 is an enhancer regulating gene transcript.