Study of an enhancer in promoter region of human colorectal cancer related ST13 gene.
- Author:
Gong-xing CHEN
1
;
Jia-wei ZHANG
;
Shu ZHENG
Author Information
- Publication Type:Journal Article
- MeSH: Base Sequence; Carrier Proteins; genetics; Colorectal Neoplasms; genetics; Enhancer Elements, Genetic; genetics; Genes, Reporter; Humans; Molecular Sequence Data; Promoter Regions, Genetic; genetics; Transcription, Genetic; Transfection; Tumor Suppressor Proteins; genetics
- From: Journal of Zhejiang University. Medical sciences 2004;33(5):385-389
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo 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.
METHODSSeveral 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.
RESULTSSeveral 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).
CONCLUSIONThe 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.
