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Target protein for Alzheimer's disease and coding gene and application thereof

An Alzheimer's disease, protein technology, applied in the field of genetic engineering, can solve problems such as unclearness and Aβ accumulation

Inactive Publication Date: 2011-09-07
HEBEI AGRICULTURAL UNIV.
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, exactly what causes Aβ accumulation and how it is linked to dementia remains unclear

Method used

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  • Target protein for Alzheimer's disease and coding gene and application thereof
  • Target protein for Alzheimer's disease and coding gene and application thereof
  • Target protein for Alzheimer's disease and coding gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Example 1: The Drosophila dZIP1 gene is responsible for Zn uptake in brain and body tissues

[0057] 1. Sequence analysis and comparison of human hZip1 ortholog gene dZip1 in Drosophila

[0058] Using NCBI Blast software, Blast the amino acid sequence of the human Zip family gene hZip1 in the GenBank flybase database. As a result, the amino acid sequence of Drosophila melanogaster (Drosophila melanogaster) dZip1 (CG9428-PA) has the highest amino acid sequence identity with hZip1 (sequence identity = 29 %, E value = 2e-15). dZip1 is located on chromosome 2 of the Drosophila genome, and the dZip1 ORF region has 1135bp (including a 76bp Intron between 1018-1094bp), encoding 352aa, and the predicted molecular weight is 37836Da. Using TMHMM software to predict the transmembrane region of the dZip1 protein structure, it is shown that dZip1 has 8 typical TM domains of the Zip family, and contains a variable region between TM3 and TM4, which is located in the cytoplasm ( fig...

Embodiment 2

[0077] Example 2: Specific regulation of the expression level of dZip1 in the Drosophila nervous system can significantly affect the deposition of Aβ amyloid in the brain

[0078] Thioflavin-S (Thioflavin S, TS) staining can specifically identify Aβ fibers, which is used in this experiment to detect the deposition of Aβ42 fibers in the Drosophila brain. The specific operation is as follows. The Drosophila brain was first fixed in 4% paraformaldehyde and 2% Triton-X100 solution for 2 hours, and then transferred to 0.25% TS solution containing 50% ethanol overnight. Then wash with 50% ethanol once for 10 minutes, and wash with PBS three times for 10 minutes each time. After that, focusclear (Pacgen Biopharmaceuticals Co.) was added dropwise, and the coverslip was covered. The slices were observed with a Zeiss LSM 510 confocal laser microscope, and data analysis was performed with LSM 510 analysis software. TS-positive deposits in mushroom body regions of the Drosophila brain w...

Embodiment 3

[0079] Example 3: Changes in Aβ amyloid deposition in Drosophila brain are closely related to Zn balance changes caused by regulation of dZip1 expression level

[0080] 1. Real-time detection of dZip1 specific overexpression in the Drosophila nervous system and Zn accumulation in the brain caused by RNAi by Zinquin staining

[0081] Since Zinquin staining can specifically label Zn ions, this experiment uses this technique to detect the effect of different expression levels of dZip1 on Zn uptake in Drosophila brain in real time. The specific operation is as follows: Drosophila after eclosion for 10 days was transferred to the culture medium containing 4mM ZnCl 2 Drosophila brains were dissected at 40h and 72h respectively, and incubated in 25μM zinquin (Sigma) solution at 37°C for 30 minutes. Then, wash with PBS solution 3 times, 5 minutes each time. The treated Drosophila brain was observed and photographed under a fluorescent microscope (Nikon, Diaphot 300). the result sho...

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Abstract

The invention relates to the field of gene engineering, in particular to a target protein for Alzheimer's disease (AD) and a coding gene and application thereof. The symptoms of patients with the AD can be obviously improved or delayed by ribonucleic acid interference (RNAi) of expression of the target protein Zip1 which can be used for treating human neurodegenerative diseases, particularly the AD. The target protein for human AD has an amino acid sequence shown as SEQ ID NO.3 or 5, and the gene has a nucleotide sequence shown as SEQ ID NO.4 or 6. Through the RNAi of the target gene, the early cognitive ability of the patients with the AD can be improved, the deposition of an amyloid beta (A beta) protein in a brain and nerve degeneration caused thereby are slowed down or depressed, athletic ability is improved and life is prolonged. The invention is expected to provide a new treatment strategy and a key target for treating the AD.

Description

technical field [0001] The present invention relates to the field of genetic engineering, in particular, the present invention relates to a target protein of Alzheimer's disease, its encoding gene and application. The present invention finds that it can be used to treat human neurodegenerative diseases, especially the target gene Zip1 of Alzheimer's disease, and interfering with the expression of Zip1 through RNAi can significantly improve or delay the symptoms of Alzheimer's patients. Background technique [0002] Alzheimer's disease (AD) is a major neurodegenerative disease affecting human health at present. Because its onset is closely related to age and mostly occurs in the elderly, the incidence rate will increase with the aging of the social population. higher. In the United States, there are 2047 AD patients for every 8 million population, and there are 6-10 million people in China. However, so far humans have no effective measures to treat the disease. [0003] β-...

Claims

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Application Information

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IPC IPC(8): C07K14/47C12N15/12C12N15/63A61K48/00A61P25/28
Inventor 郎明林范强旺王磊周兵
Owner HEBEI AGRICULTURAL UNIV.
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