Western Blot Hela Cos7 - Show nearly identical genes of two related organisms same expression (Jan/20/2006 )
Hi there, my question: How probable is it that one gene of two closely related organisms shows the same expression after transfection of Hela/Cos7 cells? The gene sequences in the two organisms show over 90% sequence homology.
Hi there,
I'm slightly unsure what exactly you mean here...
Does your question refer to the same gene from two different organisms that has been put into the same expression plasmid before being transfected?
Gene expression in cells really comes down to the promoter you use in the expression plasmid (maybe you should have a little read about promotors and expression plasmids?).
If you use the same promotor then yes, the expression should be the same as long as your transfection efficiency is the same and the sequence does not contain exotic base triplets that aren't usually used in these cells. Considering the fact that both sequences are from closely related species, the latter would have an influence on the expression of both genes.
Hope that helps
LeserattePD
Right. Thanks. I work with one particular gene which I characterize in two different species.
It turned out, that the sequences of my gene are nearly the same in the two organisms.
According to what you said, I could theoretically expect the same expression results in eukaryotic cells in case I use the same expression plasmid and promoter for transfection (what I of course do).
If there no antibodies against that protein, you are studying, yet. You should make them and then you can check the endogenous protein expression levels in tissues samples from those 2 different species.
Actually what you could do is a Northern blot. Then you can compare RNA expression levels betwwen species and more interesting between different tissues/organs.
Yeah... no reason I can think of why they shouldn't.
But you really really need to compare transfection efficiency for transient transfections and copy-number for stable transfected cells.
It turned out, that the sequences of my gene are nearly the same in the two organisms.
According to what you said, I could theoretically expect the same expression results in eukaryotic cells in case I use the same expression plasmid and promoter for transfection (what I of course do).