DNA damage - techniques (Apr/12/2007 )

hi....
I am trying to see if my protein has to do with the maintainance of DNA repair.... but i am not sure wher or how should i start testing it.
I am working with plants but I guess that animal assays can also work... Any idea will be really wellcome....I am at lost here.
I am also looking at some papers but not much come out.....



thanks in advance

Hi,

I know that this sounds an obvious question but do you know which part of DNA repair it might be involved in??

If you think it might be involved in signalling or early events in DNA repair you could try co-localisation studies. Forgive my ignorance with plants but in cultured mammalian cells co-localisation of your protein with gamma-H2AX or 53BP1 would imply that the protein may be involved in double strand break repair as these proteins both form foci at sites of DSBs. This technique is also possible with other repair pathways.

Other than that if you think your protein may involved at the nitty gritty level of DNA repair such as processing the DNA then sinple repair assays using oligonucleotides or plasmids might work with immunodepletion of your protein in cell extracts or RNAi.

Hope this helps,
Chewie

Chewie

thank you very much

I am not completely sure where my protein may be acting but, till the moment, everything indicates that is in the signaling pathway.
I will try colocalization.
thank you very much