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Method for measuring influencing factors to BAFF-R (B cell-Activating Factor-Receptor) gene promoter activity of human

A technology of influencing factors and promoters, applied in the determination/testing of microorganisms, biochemical equipment and methods, etc.

Inactive Publication Date: 2012-05-09
AFFILIATED HOSPITAL OF NANTONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no relevant report on the effect of IFN-γ and NF-κB on the expression of BAFF-R

Method used

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  • Method for measuring influencing factors to BAFF-R (B cell-Activating Factor-Receptor) gene promoter activity of human
  • Method for measuring influencing factors to BAFF-R (B cell-Activating Factor-Receptor) gene promoter activity of human
  • Method for measuring influencing factors to BAFF-R (B cell-Activating Factor-Receptor) gene promoter activity of human

Examples

Experimental program
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Embodiment Construction

[0037] A method for determining factors affecting the activity of human BAFF-R gene promoter, comprising the following steps: (1) primer design:

[0038] Use the primer design software to design eight fragment primers on the 5' flanking sequence of the BAFF-R gene in a tiling manner. Eight pairs of primers have the same downstream primers. The 3' end of the primers adds a restriction endonuclease HindIII site, and the 5' Add a restriction endonuclease Mlu I site at the end, and add 6 protective bases at both ends to facilitate digestion and insertion into the multiple cloning site of the vector; the primer sequences of the eight fragments and the downstream primer sequences are:

[0039]

[0040] Among them, 1-8 are the upstream primer sequences of eight fragments of different lengths, and 9 is the common downstream primer sequence of the 3' ends of the eight fragments; the above primers are used to amplify the corresponding products B1 (549bp), B2 (691bp), B3 (878bp) , B4(...

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Abstract

The invention discloses a method for measuring influencing factors to BAFF-R (B cell-Activating Factor-Receptor) gene promoter activity of a human. The method comprises the following steps of: primer design, cloning of BAFF-R gene 5' flanking region, construction of BAFF-R gene 5' flanking region sequence loss plasmid, cell culture, interference of an IFN-gamma and NF-kappaB inhibitor to cell proliferation, transfection of a recombinant, detection of luciferase activity, interference of the IFN-gamma and NF-kappaB inhibitor to the BAFF-R promoter activity and the mRNA expression and the like.The method is reliable and easy to operate and provides an essential basis for further studying a regulatory mechanism of the BAFF-R.

Description

Technical field: [0001] The invention relates to a method for determining factors affecting the activity of human BAFF-R gene promoter. Background technique: [0002] BAFF-R is the specific receptor of BAFF (B cell activating factor), which can specifically bind to BAFF. BAFF / BAFF-R forms a complex to regulate the self-stabilization of B cells and participate in the proliferation and function of B and T lymphocytes. adjust. Studies have confirmed that BAFF-R and its ligand BAFF mRNA and protein levels are abnormally expressed in B cell-related diseases. BAFF-R is the specific receptor of BAFF, which plays an important role in the process of BAFF's biological function, and at the same time regulates the proliferation and survival of B lymphocytes, and maintains the homeostasis of B lymphocytes' own environment. The combination of BAFF and BAFF-R can activate the classic and non-classical NF-κB signaling pathways. BAFF-R can promote the survival and proliferation of normal B...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12Q1/66C12Q1/34
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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