[Yeast] integration, but low expression

[Nicolas Brandes]

Hi,

 

Here is what I have done: I cloned my YFP gene into the pRS303 integration
vector and integrated this plasmid into the yeast chromosome at the
his3-loci. I have done this to get a stable YFP expression. But now I have
the problem that the expression level of my protein is very low. For my
further experiments I would like to have a higher expression. 

 

Can someone tell me what the expression level of my YFP gene is compared to
the expression level with a high-copy plasmid? The pRS30x series integrates
typically as a single-copy, so I guess the expression is very low.

 

And I would like to overexpress my gene. Can I somehow get the same rate of
expression with an integrated gene compared to the expression with a
high-copy plasmid? Do I need a yeast promoter to get high expression and do
I have to fuse my gene then with an endogenous yeast promoter? Or can I just
use another vector which integrates in multiple copies?

 

Thanks for your help,

Nico

 

 

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Nicolas Brandes 
University of Michigan
Department of MCDB



 

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