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Recombinant plasmid containing effective B cell activation factor gene promotor and its preparation method and uses

An activating factor, recombinant plasmid technology, applied in biochemical equipment and methods, botanical equipment and methods, gene therapy and other directions, can solve the problem of no changes in the function of BAFF gene regulatory elements, and achieve simple methods and accurate recombinant plasmids. Effect

Inactive Publication Date: 2009-02-04
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the research on the precise location of the upstream regulatory elements of the BAFF gene and its interaction with the corresponding DNA-binding protein is still blank, and there is no report on the functional changes of the BAFF gene regulatory elements under pathological conditions.

Method used

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  • Recombinant plasmid containing effective B cell activation factor gene promotor and its preparation method and uses
  • Recombinant plasmid containing effective B cell activation factor gene promotor and its preparation method and uses
  • Recombinant plasmid containing effective B cell activation factor gene promotor and its preparation method and uses

Examples

Experimental program
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Effect test

Embodiment 1

[0018] Example 1: Preparation of recombinant plasmid containing effective BAFF gene promoter

[0019] The promoter of the human BAFF gene is located in the region of about 1020 bp in the upstream 5' flanking region of the gene (see Kawasaki A, Tsuchiya N, Fukazawa T, et al. Analysis on the association of human BLYS (BAFF, TNFSF13B) polymorphisms with systemic lupus erythematosus and rheumatoid arthritis[ J]. Genes Immun, 2002, 3(7): 424-429), see Figure 4 . The purpose of this study is to analyze the promoter regulation activity of a partial fragment (-1349~-743bp sequence) of the upstream 5'flanking region of the human BAFF gene. The sequence and the reporter gene are combined into a recombinant promoter plasmid, and the promoter activity of the recombinant is observed through gene transfection and the activity of the reporter gene, which lays the foundation for exploring the regulatory target sequence that activates / blocks the BAFF gene at the transcriptional level.

[00...

Embodiment 2

[0128] Example 2: Effects of some cytokines on the transcriptional regulation of BAFF

[0129] A large number of studies have shown that the abnormal expression of BAFF is closely related to the immunopathology of SLE and other autoimmune diseases. Foreign literature reports that the expression of BAFF molecules in myeloid cells such as monocytes, macrophages, and dendritic cells is affected by various cytokines such as IL-10, IFN-γ, and IL-4. Upregulation / downregulation of BAFF protein synthesis and secretion. However, how these cytokines regulate the expression of BAFF molecules in myeloid cells and whether they affect the synthesis and secretion of BAFF at the transcriptional level has not yet been clearly explained. In this study, the recombinant phBAFF3 constructed in "Example 1" and containing a sequence of -1349 to -743bp with strong transcriptional activity was used as the experimental platform, and it was transiently transfected into human myeloid leukemia cells HL-6...

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Abstract

The invention discloses a recombination plasmid with effective BAFF gene promotor in the biomedical engineering domain, which is characterized by the following: the recombination plasmid is composed of -1349--743bp fragment of human BAFF gene upstream promotor sequence and alficetin acetylase(CAT); The usage and method are provided for preparing recombination plasmid; the recombination plasmid is exact and reliable; it's easy to activate and block-up controlling target dots of BAFF gene in transcription horizon.

Description

technical field [0001] The invention relates to the technical field of biomedical engineering, in particular to a recombinant plasmid containing an effective B cell activating factor gene promoter, a preparation method and application thereof. Background technique [0002] Systemic lupus erythematosus (SLE) is a common and frequently-occurring disease, mainly manifested by the excessive proliferation and activation of B cells and the production of various autoantibodies (anti-double-stranded DNA antibodies are the most important). The disease mostly occurs in young and middle-aged women aged 20 to 50, with a male to female ratio of about 1:8; if the diagnosis and treatment are not timely, 60 to 70% of patients will suffer from renal failure, seriously affecting the quality of labor. However, the pathogenesis of SLE is still unclear, so far there is no specific diagnostic method, let alone ideal treatment. Among the many possible pathogenesis of SLE, the theory of excessive ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/63C12N15/54C12N15/12C12N15/28A61K48/00A61P37/02
Inventor 仲人前周琳孔宪涛高春芳王皓屠小卿陆慧琦
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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