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Transcription factors that dont bind DNA


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8 replies to this topic

#1 cardosopedro

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Posted 14 January 2011 - 08:08 AM

Hi.

I am studying a protein which is probably a transcription factor (it encodes a transcription factor domain although this domain is not involved in DNA-binding). Bioinformatics predictions show that apparently the protein does not have DNA-binding motifs, although I have functional data highly suggestive of the protein being involved in gene expression regulation.

Is it possible that it is a non-binding transcription factor? Are these proteins common? Do they have particular features? Could it be interacting with a DNA-binding molecule allowing the regulation of gene expression? Does anyone know how I can predict this and look for the putative mediator(s)?


A big thanks for anyone who can help!
;)

Edited by cardosopedro, 14 January 2011 - 08:09 AM.


#2 cardosopedro

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Posted 21 January 2011 - 02:14 AM

Can someone help me please?
:)

#3 bob1

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Posted 22 January 2011 - 01:01 AM

Well there are the E2F group and Rb that don't bind to DNA but do control transcription at a certain level. You could do ChIP to identify if it is binding DNA and which sequence it is binding to.

#4 cardosopedro

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Posted 22 January 2011 - 02:41 AM

Thank you.

Yes, I thought of that. But there is not comercially available antibody against my protein. I will have to produce my own antibody.

#5 Radish

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Posted 25 January 2011 - 01:43 PM

I would proppose that you cloned your protein from total RNA, add a tag (flag or myc) get into a vector (gateway system would probably be the best.
And in 2 to 3 months you could overexpress it in your target cells and use a flag or myc anti-body to do the pull-down.


best regards

Radish

#6 cardosopedro

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Posted 26 January 2011 - 02:10 AM

Very nice idea, thanks!

#7 Radish

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Posted 26 January 2011 - 11:46 AM

No problem.
Pm me if you need any help!

#8 gaurav amit prakash

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Posted 31 January 2011 - 07:23 AM

hey

as we all know that there are two type of tfs ;
1) gtf
2) taf
so first of all u have to see that they are working as gtfs or tafs
for gtfs u have to use chip to see that either they are binding to dna or not

for tafs u have to see it by south western blotting.

check it it may say something..... :ph34r: <_< :rolleyes: :) B)

#9 Radish

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Posted 31 January 2011 - 12:39 PM

This may sound stupid but...
what are tafs and gtfs?


((TBP)-associated factors and General transcription factors??)

if so you don't have limited techniques for any particular class...




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