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Deinococcus radiodurans transcription inhibition factor

A technology of Deinococcus radiodurans, transcription inhibitor, applied in bacterial peptides, chemical instruments and methods, peptide sources, etc., can solve the problems of DdrO function and regulation mode and objects that have never been studied and reported.

Inactive Publication Date: 2014-12-17
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But so far, the specific function, regulation mode and object of DdrO have never been reported by research

Method used

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  • Deinococcus radiodurans transcription inhibition factor
  • Deinococcus radiodurans transcription inhibition factor
  • Deinococcus radiodurans transcription inhibition factor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] (1) DdrO binds to gene promoters containing RDRM sites in vitro

[0031] DdrO and dr0070 , dr0099 , dra0151 , dr0219 , dr0326 , dra0346 , dr0423 , dr0596 , dr0906 and dr1039 The full promoter of the isogen was added in binding buffer (100 mM NaCl, 20 mM Tris-HCl 8.0, 5 mM MgCl 2 ) for 40 minutes. Use 12% TB-PAGE gel to detect the migration of DNA bands. Experiments have shown that the promoters of the above genes can all combine with DdrO, and the migration of the bands occurs ( figure 1 ).

[0032] (2) The RDRM site of the gene promoter is required for DdrO binding in vitro

[0033] DdrO and P without RDRM sites dr0070- ,P dr0099- ,P dr2338- and P dr0423- Isogenic promoters in binding buffer (100 mM NaCl, 20 mM Tris-HCl 8.0, 5 mM MgCl 2 ) for 40 minutes. Use 12% TB-PAGE gel to detect the migration of DNA bands. Experiments have shown that none of the above-mentioned gene promoters without RDRM sites can bind DdrO, and no band migration occurs (...

Embodiment 2

[0039] (1) DdrO binds to gene promoters containing RDRM sites in vitro

[0040] DdrO and dr1143 , dr1289 ,dr1696 , dr1771 , dr1775 , dr1913 , dr1921 , dr2256 , dr2275 , dr2336 and dr2574 The full promoter of the isogen was added in binding buffer (150 mM NaCl, 30 mM Tris-HCl 8.0, 5 mM MgCl 2 ) for 40 minutes. Use 12% TB-PAGE gel to detect the migration of DNA bands. Experiments have shown that the promoters of the above genes can all combine with DdrO, and band migration occurs ( figure 1 ).

[0041] (2) The RDRM site of the gene promoter is required for DdrO binding in vitro

[0042] DdrO and P without RDRM sites dr0326- ,P dra0346- and P dr2574- and other promoters in binding buffer (150 mM NaCl, 30 mM Tris-HCl 8.0, 5 mM MgCl 2 ) for 40 minutes. Use 12% TB-PAGE gel to detect the migration of DNA bands. Experiments have shown that none of the above gene promoters can bind DdrO, and no band migration occurs ( figure 2 ).

[0043] (3) DdrO binds DNA dam...

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Abstract

The invention discloses a Deinococcus radiodurans transcription inhibition factor. The transcription inhibition factor can be combined with an RDRM site-containing DNA damage emergency response and repair gene promoter in Deinococcus radiodurans in vitro. Components of a combination reaction buffer comprise 100-200mM of NaCl, 10-50mM of Tris-HCl 8.0 and 5-10mM of MgCl2, and the reaction temperature is 30DEG C. The transcription inhibition factor can inhibit the transcription expression of the DNA damage emergency response and repair gene in vivo. The transcription inhibition factor has not been reported so far is disclosed in the invention, and is of great significance for researching DNA damage repair.

Description

technical field [0001] The invention relates to a Deinococcus radiodurans transcriptional inhibitor DdrO, relates to a DNA sequence capable of specifically binding to DdrO and its sequence characteristics, and performs the function of an inhibitory factor in vivo. Background technique [0002] In molecular biology, a transcription factor refers to a protein that can bind to a specific DNA sequence upstream of a gene and can regulate the transcription of the gene. It can regulate the combination of RNA polymerase and DNA template, and can also regulate the function of RNA transcription of RNA polymerase on DNA template. Transcription factors not only bind to the promoter region upstream of genes, but also form transcription factor complexes with other transcription factors to affect the transcription of genes. [0003] Transcription factors and their systems have been widely used in molecular biology, molecular medicine and other fields. Using transcription factor systems, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/195
CPCC07K14/195
Inventor 华跃进王云光王梁燕
Owner ZHEJIANG UNIV
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