gfp fusion proteins - is liker needed if so how many amino acids usual choice? (Jan/29/2007 )

hello all

i am planning to clone GFP to my gene and get 5'end fusions and 3'end fusions, and try to get them expressed in plants, view thru confocal or under UV and finally do western

for fusion proteins is a long linker (20-50 aminoacids ) needed for proper folding and expression?

and is it ok if i skip stop codon of the first gene but keep the start codon of the second ?

i read some papers and some people only used 6-7 aminoacids for gfp fusions? any suggestions?


also in case of fusion protiens is it usual to skip the start codon of the second protein ?

any thoughts appreciated

regards

laxmi

When we make fusion protiens, we have 2 glycine between the 2 genes. So far this has worked.

You should remove the first stop codon and having the second start codon is not necessary. You could have the second start if you wanted.

Good Luck !!!

thank you very much scolix i am going to design primers to do away with the stop codon in the first and keep the start codon in the second and see.and some glycine residues invbetween so that i can have a kpnI site inbetween

thanks again

regards
laxmi