Protein forming complex with c-fos protein, nucleic acid encoding the same and method of using the same

a technology of protein forming complex and c-fos protein, which is applied in the field of protein, can solve the problems of enormous time and labor required to achieve 1:1 molecule analysis technique and the use of two hybrid methods

Inactive Publication Date: 2006-08-17
KEIO UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0031] An object of the present invention is to provide a complex that interacts with the c-Fos protein, which is well known as a transcription control factor, as a target protein.

Problems solved by technology

However, such a thorough 1:1 molecule analysis technique as the two-hybrid method takes enormous time and labor.

Method used

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  • Protein forming complex with c-fos protein, nucleic acid encoding the same and method of using the same
  • Protein forming complex with c-fos protein, nucleic acid encoding the same and method of using the same
  • Protein forming complex with c-fos protein, nucleic acid encoding the same and method of using the same

Examples

Experimental program
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Effect test

example 1

[0367] Cotranslation selection / screening of IVV was carried out by using the c-Fos protein as a bait and a mouse brain cDNA library as a prey (FIG. 2), and as a result, genes or nucleic acid sequences encoding novel proteins that can form a complex with the c-Fos protein were obtained.

[0368] The preparation method of the bait, c-Fos protein, was as follows. A DNA template was prepared from a pCMV-FosCBPzz vector (SEQ ID NO: 168) by PCR (primers 5′ SP6(O29)T7-FosCBPzz (SEQ ID NO: 169) and 3′ FosCBPzz (SEQ ID NO: 170), and PCR program CYCB1 (refer to Table 1)) using TaKaRa Ex Taq (Takara Shuzo). The DNA template was transcribed (37° C., 2 hours) by using RiboMAX™ Large Scale RNA Production Systems (Promega) to prepare a mRNA template of the bait c-Fos protein. A bait DNA made to coexist was prepared by PCR (primers 5′ DNA (SEQ ID NO: 172) and 3′ DNA (SEQ ID NO: 173)) using DNA-Fos / Jun (SEQ ID NO: 171) containing the Fos / Jun binding sequence as a template according to the PCR program ...

example 2

[0374] A prey IVV library was prepared from the bait c-Fos and a mouse brain cDNA library in the same manner as that used in Example 1, and the screening / selection procedure (FIG. 2) was also performed in the same manner as that used in Example 1. However, in this example, the first stage selection using IgG beads in the two-stage screening was repeated 3 times, and the two-stage selection was performed only for the 4th time to obtain the proteins of SEQ ID NOS: 47 to 56 (amino acid sequences of Fip-cx.1), SEQ ID NOS: 57 to 76 (amino acid sequences of Fip-cx.2), SEQ ID NOS: 77 to 81 (amino acid sequences of Optin), SEQ ID NOS: 82 to 84 (amino acid sequences of Snap19), SEQ ID NOS: 85 and 86 (amino acid sequences of C130020M04Rik), SEQ ID NOS: 87 to 89 (amino acid sequences of FLJ32000), SEQ ID NOS: 90 and 91 (amino acid sequences of Rit2), SEQ ID NOS: 92 and 93 (amino acid sequences of cytocrome b), SEQ ID NOS: 94 and 95 (amino acid sequences of Apoe), SEQ ID NOS: 96 and 97 (amino a...

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Abstract

Proteins that interact with c-Fos, nucleic acids encoding them and inhibitors utilizing them as well as methods for detecting an interaction and screening methods utilizing a protein that interacts with c-Fos are provided. Comprehensive analysis of transcription control factor complexes in a mouse brain cDNA library with c-Fos as a bait by using the cotranslation selection and screening of in vitro virus (IVV) and the C-terminal labeling method are conducted, thereby to analyze proteins unknown so far, proteins known so far, but unknown to form a complex with the c-Fos protein, and so forth.

Description

TECHNICAL FIELD [0001] The present invention relates to proteins that interact with c-Fos, nucleic acids encoding them and inhibitors utilizing them as well as methods for detecting an interaction and screening methods utilizing a protein that interacts with c-Fos. [0002] At present, genomic nucleotide sequences of various organisms are going to be elucidated. In researches of genomic sequences, there are expected, as post-sequencing researches of the second act, researches of analyzing meanings of the elucidated genomic information, i.e., structural and functional analyses of genes and proteins (Non-patent documents 1 and 2), analyses of protein / protein and protein / nucleic acid interactions (Non-patent documents 3 and 4) and so forth. [0003] On the basis of analyses of networks of interactions between protein and protein, protein and nucleic acid and so forth in post-sequencing genomic functional analyses utilizing such techniques as described above, it is expected to create drugs ...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): C07K14/705C12Q1/68G01N33/53C07H21/04C07K14/47C12N15/10
CPCC07K14/47C12N15/1055G01N2500/04
Inventor MIYAMOTO, ETSUKOISHIZAKA, MASAMICHIYANAGAWA, HIROSHI
Owner KEIO UNIV
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