Cloning and function analysis of 5′ end flanking sequence of EOLA1 gene
- VernacularTitle:EOLA1基因5′侧翼区的克隆及调控功能初步研究
- Author:
Ziwen LIANG
;
Zhongcheng YANG
;
Jian CHEN
;
Yu CHEN
- Publication Type:Journal Article
- Keywords:
endothelial-overexpressed lipopolysaccharide-associated factor 1 gene;
promoter;
reporter gene;
activity assay
- From:Journal of Third Military Medical University
2003;0(10):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct the ?-gal reporter genes containing the 5′-end flanking of endothelial-overexpressed lipopolysaccharide-associated factor 1 (EOLA1) gene in different sequence lengths and identify the sequence, which regulates the gene expression of EOLA1 by the ?-gal analysis system. Methods The target sequences were amplified by the method of genome walker, and were inserted into the upstream of ?-gal gene located in the ?-gal enhancer vector by the directional clone technique respectively; the regulative sequence was identified by analyzing the ?-gal activities of reconstructed plasmid in ECV304 cells. Results The regions, containing 2 659 bp and 1 951 bp upstreaming from exon 1, significantly stimulated the reporter gene activity as compared with that of the ?-gal control vector in transfected cells. But the region, containing 361 bp upstreaming from exon 1, did not stimulate the reporter gene activity. Conclusion There is an up-regulative element of gene transcription in the region of -361 to -1 951 bp in EOLA1 gene upstream.
