Bicistronic vector vs "normal" vector - (Dec/05/2010 )
Dear all,
I hope someone is able to answer my question.
What is the difference or advantage of a bicistronic vector (EGFP as a marker)compared to a normal vector expressing my protein of interest as a fusion protein (e.g. EGFP). In both cases I see a fluorescent signal after a successful transfection, right? And what about the bidirectional vector. I am a bit confused.
Every comment is very welcome.
Thank you very much
Purzel
Using a fusion with GFP may change your protein's activity or localisation. You will need to carefully test this. Using bicistronic vectors, IRES or 2A based, avoids this problem, because you'll have your native protein expressed.
rsm
Bear in mind that when using a bicistronic construct with IRES, the expression of your secondary transgene is markedly lower than the primary.
elpollodiablo on Tue Dec 7 08:07:10 2010 said:
Bear in mind that when using a bicistronic construct with IRES, the expression of your secondary transgene is markedly lower than the primary.
And why?
elpollodiablo on Tue Dec 7 08:07:10 2010 said:
Bear in mind that when using a bicistronic construct with IRES, the expression of your secondary transgene is markedly lower than the primary.
And why?
elpollodiablo on Tue Dec 7 08:07:10 2010 said:
Bear in mind that when using a bicistronic construct with IRES, the expression of your secondary transgene is markedly lower than the primary.
I am not convinced
.That is actually an area of intense discussion. Well, depends on who you're talking to, I guess. But I had recently a good discussion about it...
Here a good read about this topic:
http://www.ncbi.nlm.nih.gov/pubmed/15934839
I can recommend 2A peptides, for some circumstances.
rsm