Dear Jaballi,

in most cases, linker sequences of 10-20 amino acids are sufficient to allow both proteins to work properly.

If you fear your protein looses some of its properties, I would construct both an N-Terminal and C-terminal construct.

For the Sequence, try to use small amino acids (Gly, Ser, Ala), as this makes the linker flexible.

Also think of Codon-optimization of the linker sequencse for yeast expression

Hope that helps

Best and good luck

Chris

Dear Jaballi,

in most cases, linker sequences of 10-20 amino acids are sufficient to allow both proteins to work properly.

If you fear your protein looses some of its properties, I would construct both an N-Terminal and C-terminal construct.

For the Sequence, try to use small amino acids (Gly, Ser, Ala), as this makes the linker flexible.

Also think of Codon-optimization of the linker sequencse for yeast expression

Hope that helps

Best and good luck

Chris

I agree with Christian as you can use any small amino acids as a linker. Most prepered is poly gly or poly ala as they allow free movement and doesnot alter folding patter of the domain connected. You can use a mix of small aminoacids also. The size of the linker can be 5-12 amino acids. If possible go through this paper it will help you.

http://www.ncbi.nlm.nih.gov/pubmed/23026637

You could try a (G4S)-Linker. Usually this is long enough and provides enough flexibility.