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Method for detecting activity of human BAFF-R gene promoter

A measurement method and a promoter technology, which are applied in the determination/inspection of microorganisms, biochemical equipment and methods, etc., can solve the problems that have not yet been reported in the research, and achieve the effect of easy operation

Inactive Publication Date: 2010-07-21
AFFILIATED HOSPITAL OF NANTONG UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are many studies on the expression level of BAFF-R and the occurrence and development of diseases, but there is no relevant report on the research on the transcriptional regulation of BAFF-R gene

Method used

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  • Method for detecting activity of human BAFF-R gene promoter
  • Method for detecting activity of human BAFF-R gene promoter
  • Method for detecting activity of human BAFF-R gene promoter

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Embodiment Construction

[0032] A method for measuring the activity of human BAFF-R gene promoter, which is characterized in that it comprises the following steps:

[0033] material:

[0034] Strain DH5α is the preserved strain of this unit. Human Burkitt’s lymphoma cell line Raji was donated by the Immunology Laboratory of Nantong University, and the multiple myeloma cell line KM3 was donated by Shanghai Second Military Medical University. Mammalian Genomic DNA Extraction Kit, Gel Recovery Kit (Biyuntian); PCR reagents, DNA Marker (Dalian Bao Bio-Takara Company); pGL3-Basic vector, pGL3-Control plasmid, internal reference plasmid psv-β-gal plasmid, Luciferase reporter gene detection kit, T4 ligase, restriction enzymes Mlu I, HindIII, plasmid extraction kit, purchased from Promega; fetal calf serum, RPMI1640 medium was purchased from Gibco. The liposome transfection reagent is a product of Invitrogen, USA;

[0035] (1) Primer design:

[0036] Use primer design software Primer premier5.0 to design eight frag...

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Abstract

The invention discloses a method for detecting the activity of a human BAFF-R gene promoter, which comprises the following steps: designing a primer, cloning a BAFF-R gene 5' flanking region, constructing a sequence-loss plasmid of the BAFF-R gene 5' flanking region, culturing cells, recombining a report gene plasmid transiently transfected cell, detecting luciferase activity and the like. The method is easy to operate and accurate, detects the difference between transcriptional activities of a BAFF-R gene 5' upstream sequence in different cells and in different areas, determines the area where a core promoter of a BAFF-R gene is, and lays a foundation for studying the effect of a BAFF-R gene transcriptional regulation element and clarifying the expression regulation mechanism of the gene.

Description

Technical field: [0001] The invention relates to a method for measuring the activity of human BAFF-R gene promoter. Background technique: [0002] BAFF-R is a member of the tumor necrosis factor receptor family, and is a specific receptor for B lymphocyte stimulator (BLyS), also known as (B cell activating factors belonging to the TNF family, BAFF). BLyS is involved in the regulation of the proliferation and function of B and T lymphocytes by binding to three receptors: transmembrane protein activator (TACI), B cell maturation antigen (BCMA) and BAFF receptor (BAFF-R). Overexpression of BLyS will make B lymphocytes continue to proliferate and secrete more autoantibodies, leading to the occurrence of a series of autoimmune diseases, and it will also make lymphocytes proliferate out of control and cause the occurrence of B cell malignancies. Among the three receptors of BLyS, BAFF-R is its specific receptor that can specifically bind to it. BAFF-R is necessary for the survival and...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/66
Inventor 鞠少卿王惠民王跃国袁宏香丛辉王旭东丁伟峰浦江倪红兵
Owner AFFILIATED HOSPITAL OF NANTONG UNIV
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