[Study of an enhancer in promoter region of human colorectal cancer related ST13 gene].
Gong-xing Chen, Jiawei Zhang, Shu Sen Zheng
Abstract
Gong-xing Chen, Jiawei Zhang, Shu Sen Zheng
Abstract
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.
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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.
Key concepts: Enhancer, Luciferase, Gene, Biology, Molecular biology, Transfection, Regulatory sequence, Promoter