[Identification and clone of human Alzheimer's disease related gene nicastrin promoter].
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.


CONCLUSIONS
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.
Yang
Mei
M
National Key Lab of Medical Genetics of China, Central South University, Changsha 410078, China. nlmglcy@xysm.net
Cai
Fang
F
Wang
Rui-shan
RS
Pan
Qian
Q
Long
Zhi-gao
ZG
Deng
Xiao-yun
XY
Xia
Kun
K
Xia
Jia-hui
JH
Zhang
Zhuo-hua
ZH
D000544:Alzheimer Disease / Q000235:genetics*; D053829:Amyloid Precursor Protein Secretases; D003001:Cloning, Molecular; D017930:Genes, Reporter / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D008562:Membrane Glycoproteins / Q000032:analysis / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000235:genetics*
D008562:Membrane Glycoproteins; C413604:nicastrin protein; D053829:Amyloid Precursor Protein Secretases