Purpose of PolyA in cloning (View forum version)



redo

Posted 07 May 2014 - 07:28 PM

Hi everyone,
I had a beginner's question, new to all of this. While looking at plasmid constructs, I notice sometimes, when 2 proteins need to be co-expressed, they are cloned in as:

Protein A - IRES - Protein B

 

but other times as:

Protein A - IRES - Protein B - PolyA

What is the purpose of the poly A sequence? Does it boost up protein levels by stabilizing the mRNA? 

Thanks a lot!


Trof

Posted 10 May 2014 - 10:33 AM

Are you sure there is no polyA signal in the other construct? That's a place where polymerase detaches. I can't imagine what would be effect of ommiting it, trascripts stopping unexpectedly in different parts of construct?

PolyA is also one of the marks of eukyrote mRNA, I would even expect it to have other functions apart from mRNA stabilization.

 

Is't there a possibility that there is some non-standard polyA signal instead?