eGFP Expression Efficiency

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pmngrad
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eGFP Expression Efficiency

Post by pmngrad » Thu Apr 22, 2010 5:30 pm

Hi All,

Say you have a transgenic mouse who has been genetically modified to express eGFP where ever Protein X is expressed. What events might cause eGFP not to be expressed in a cell, when you are certain that Protein X is expressed. For example, say you are looking at two cells fluorescently stained...

1) Positive for eGFP and Positive for anti-Protein X
2) Negative for eGFP and Positive for anti-Protein X

I suppose that if the GFP protein was mis-folded, it would prevent the eGFP signal, however I would not expect this to happen too often. Any other ideas?

Thank you :?

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JackBean
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Post by JackBean » Fri Apr 23, 2010 8:08 am

I don't think, that GFP misfolding would occur. You could have some other substance, which would adsorb the emitted light, that's used sometimes, but for protein-protein interaction research. But I don't think, that something like that occurred in the cell naturally :)

What about some silencing?
http://www.biolib.cz/en/main/

Cis or trans? That's what matters.

pmngrad
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Re: eGFP Expression Efficiency

Post by pmngrad » Fri Apr 23, 2010 2:25 pm

Thanks for that response - I'll look into what silencing can do to play into this.

I think that the actual terms I should have used for this question is:

What would cause reduced eGFP fidelity in some cells and not others. I'm not sure if that key word triggers anyone's brains! Thanks again!

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JackBean
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Post by JackBean » Fri Apr 23, 2010 4:46 pm

well, the silencing is not the same in all cells
http://www.biolib.cz/en/main/

Cis or trans? That's what matters.

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