Identification and clone of human Alzheimer's disease related gene nicastrin promoter.
- Author:
Mei YANG
1
;
Fang CAI
;
Rui-shan WANG
;
Qian PAN
;
Zhi-gao LONG
;
Xiao-yun DENG
;
Kun XIA
;
Jia-hui XIA
;
Zhuo-hua ZHANG
Author Information
1. National Key Lab of Medical Genetics of China, Central South University, Changsha 410078, China. nlmglcy@xysm.net
- Publication Type:Journal Article
- MeSH:
Alzheimer Disease;
genetics;
Amyloid Precursor Protein Secretases;
Cloning, Molecular;
Genes, Reporter;
genetics;
HeLa Cells;
Humans;
Membrane Glycoproteins;
analysis;
genetics;
Promoter Regions, Genetic;
genetics
- From:
Journal of Central South University(Medical Sciences)
2006;31(1):9-13
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To identify the promoter of human nicastrin (NCT) gene, a major component of gamma-secretase which is closely related with pathogenesis of Alzheimer's disease.
METHODS:Promoter of human Alzheimer's disease related gene, nicastrin, a 1768 bp fragment was firstly isolated from human genomic DNA by PCR. This fragment's 3 flanking end was 4 bp upstream to the start codon ATG (+1) of the gene. This fragment was used as template, a series of deleted fragments were amplified and constructed to the pGL3-Enhancer plasmid with the artificial designed linkers. The relative activity of their promoter in Hela cells was studied by dual-luciferase assay.
RESULTS:The 420 bp fragment showed the strongest activity, and the 237 bp fragment was the minimal fragment in length with activity.
CONCLUSION:The promoter of NCT is located at -432/-133 region upstream the translational start codon, while its basal promoter is between -359/-90 that drives the transcription of reporter gene in Hela cells.
