Transcriptional promoter for human BACE1 gene as well as main cis-acting elements and application of transcriptional promoter

A gene transcription and promoter technology, applied in the direction of recombinant DNA technology, DNA / RNA fragments, medical preparations containing active ingredients, etc., can solve few problems

Active Publication Date: 2015-02-18
CHILDRENS HOSPITAL OF CHONGQING MEDICAL UNIV
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Problems solved by technology

The cis-acting elements bound by the above-mentioned many transcription factors are located far upstream from the transcription initiation site and play a regulatory role in transcription, and the transcription factors that combine with these cis-acting elements to play a transcriptional regulatory role depend on It plays a role in the interaction with general transcription fac

Method used

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  • Transcriptional promoter for human BACE1 gene as well as main cis-acting elements and application of transcriptional promoter
  • Transcriptional promoter for human BACE1 gene as well as main cis-acting elements and application of transcriptional promoter
  • Transcriptional promoter for human BACE1 gene as well as main cis-acting elements and application of transcriptional promoter

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Experimental program
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Embodiment

[0025] 1. Materials and methods

[0026] 1.1 Search the GenBank database and literature to determine the promoter region of the BACE1 gene.

[0027] 1.2 Deletion plasmid construction

[0028] The BACE1 promoter luciferase reporter gene p3BU-1273 / -400 contains the sequence from -1273 to -400 upstream of the translation initiation site of the BACE1 gene. The target fragment is amplified by PCR and named as B1U-1273 / -400 or B1U1273-400 (the following names are all the same as this method, B1U-1273 / -400 represents the sequence from the 1273rd nucleotide to the 400th nucleotide upstream of the translation initiation site of BACE1 gene), the primer is B1U1273F / B1U400R (see Table 1), the template is bacterial artificial chromosome DNA (RP11-970G21) (from BAC DNA library). The LA TAQ enzyme from TAKARA company was used for amplification. The reaction program was initial denaturation at 95°C for 30s, followed by 30 cycles at 98°C for 30s, 68°C for 2min, and extension at 72°C for 10mi...

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Abstract

The invention provides a transcriptional promoter for the human BACE1 gene as well as a core promoter region and cis-acting elements of the transcriptional promoter. According to the invention, the smallest promoter region defining the high-level transcription of the human BACE1 gene and the core promoter region defining basal transcription of the human BACE1 gene are firstly confirmed, and the two cis-acting elements capable of enhancing gene transcription are confirmed, the structure of the BACE1 gene promoter core region playing a key function on the transcription of the BACE1 gene is disclosed, and a transcription regulatory mechanism of the BACE1 gene and influence, found through the transcription regulatory mechanism, on the transcription and expression of the BACE1 gene are understood more deeply. Effective action target spots are provided for preventing, diagnosing and treating AD and other diseases.

Description

technical field [0001] The invention relates to a human gene promoter and its main cis-acting element, in particular to the expression and regulation of human BACE1 gene. Background technique [0002] The BACE1 gene encodes β-site amyloid precursor protein cleavage enzyme (BACE1, also known as Asp2 or memapsin2), which is a transmembrane aspartic acid protease with 501 amino acids. BACE1 is the only β-secretase in the human body, which generates β-amyloid protein (amyloid β peptide, Aβ) by cleaving amyloid precursor protein (AβPP or APP), which is a rate-limiting step of Aβ production. In the brains of Alzheimer's disease (AD) patients, Aβ is an important pathological manifestation - the main component of amyloid neuritic plaques, and Aβ deposition plays a key role in the occurrence and development of AD. Reducing the generation of Aβ by inhibiting the activity of BACE1 has become the target of drug development for the prevention and treatment of AD. [0003] The human BAC...

Claims

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Application Information

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IPC IPC(8): C12N15/113C12Q1/68A61K45/00A61P25/28
Inventor 周卫辉
Owner CHILDRENS HOSPITAL OF CHONGQING MEDICAL UNIV
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