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Recombinant plasmids containing B cell activation factor gene promoters with different lengths and their preparation method and application

A technology of activating factors and recombinant plasmids, applied in biochemical equipment and methods, botany equipment and methods, gene therapy, etc., can solve problems such as abnormal expression of downstream proteins, lack of BAFF gene promoter structure, functional analysis and transcriptional regulation , to achieve the effect of a simple method

Inactive Publication Date: 2009-10-28
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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Problems solved by technology

[0005] However, in recent years, the research progress and trend of BAFF at home and abroad have mainly focused on exploring the relationship between BAFF and SLE immunopathology at the protein and nucleic acid levels, and there is obviously a lack of research on the structure, function analysis and transcriptional regulation of the BAFF gene promoter. Research
The promoter is necessary for gene expression, it determines the space, time and expression intensity of exogenous gene expression, etc. Any structural abnormality at the promoter level or changes in transcriptional regulatory elements will directly affect the mRNA transcription. Abnormal, resulting in abnormal expression of downstream protein levels

Method used

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  • Recombinant plasmids containing B cell activation factor gene promoters with different lengths and their preparation method and application
  • Recombinant plasmids containing B cell activation factor gene promoters with different lengths and their preparation method and application
  • Recombinant plasmids containing B cell activation factor gene promoters with different lengths and their preparation method and application

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Embodiment 1

[0023] Example 1: Preparation of recombinant plasmids containing different length BAFF gene promoters and their activity detection

[0024] Foreign studies have reported that the human BAFF gene is located on chromosome 13q34, including a coding region containing 285aa, a promoter region of about 1020 bp in the upstream 5' flanking region, two non-coding regions at the 3' and 5' ends, and six exons. . Moreover, there are four polymorphic sites in the BAFF promoter region, which are respectively located at -1283G>A, -871C>T, -514T>C and -353G>C (see Figure 4 ). Accordingly, the present invention analyzes the transcriptional activity of the 1020 bp upstream regulatory sequence of the human BAFF gene segment by segment. The present invention uses computer software (DNA assist1.0) to carry out the simulation analysis of the main nucleoprotein binding to the 1020bp human BAFF gene promoter sequence (Accession: AB730224) reported by Genebank, suggesting that there are 6 Sp-1 (-32...

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Abstract

The invention relates to the technical field of biomedical engineering. At present, the structure, function analysis and transcriptional regulation of the B cell activating factor (BAFF) gene promoter are still blank. The present invention provides five recombinant plasmids containing different lengths of the B cell activating factor (BAFF) gene promoter. It consists of -1349~-1099bp, -1349~-893bp, -1349~-743bp, -1349~-431bp, -1349~-329bp fragments of the upstream promoter sequence of human BAFF gene and without promoter, but containing chloramphenicol acetyl The carrier composition of the base transferase CAT reporter gene, the invention also provides the preparation method and application of the recombinant plasmid. The recombinant plasmids constructed by the present invention containing BAFF gene promoters of different lengths are accurate and reliable, and have laid a foundation for further research on the transcriptional regulation mechanism of human BAFF gene activation; and the method of comparing the transcriptional activity of recombinant promoters with reporter gene activity is simple and can be quantified , is a simple method for screening drugs for antagonism / treatment of SLE at the transcriptional level, and also provides a new target for exploring the prevention and treatment of SLE.

Description

technical field [0001] The invention relates to the technical field of biomedical engineering, in particular to five recombinant plasmids containing different lengths of B cell activation factor (BAFF) gene promoters and their preparation methods and applications. Background technique [0002] Systemic lupus erythematosus (SLE) is a common and frequently-occurring disease, mainly manifested in the excessive proliferation and activation of B cells, the production of various autoantibodies such as anti-double-stranded DNA antibodies, and through immune complexes and other ways, causing Almost every system and organ in the body is affected. The disease mostly occurs in young and middle-aged women aged 20 to 50. According to reports, the prevalence of SLE in my country is close to 1‰, and the total number of patients in the country is as high as 1 million. If the diagnosis and treatment are not timely, 70-80% of SLE patients will suffer from renal failure, seriously affecting t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/63C12N15/65C12N15/12A61K48/00A61P37/02
Inventor 周琳仲人前孔宪涛王皓屠小卿朱烨郝婉莹
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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