two additional amino acids in gene - (Feb/07/2007 )
Hi all,
I am cloning mRFP gene to hydrophilic loop of a membrane protein. Unfortunately I have added two additional amino acids just before the mRFP gene. Now the sequence would be ATG TGT ATG...... first ATG (Met)and TGT(Cys) are additional. Could anyone of u please tell how these 2 additional AA's will affect the expression of mRFP as well as my membrane protein??? Thanks in advance....
Translation would start from the first ATG. As far as expression of mRFP is concerned, One cannot really predict it becasue its a fusion protein, you have to transfect cells and verify it works as you wished or not. At times fusion protiens are cleaved or aggregate due to the presence of fluorescence protien. Basically, its any ones guess.
thank u very much acolix....
what a nice research, inserting a fluorescent protein in a membrane protein.......I want to try it for a long time,
but, well, will the folding of the membrane protein be easily affected ? and it will be expressed in bacteria or eukaryotic ??
but, well, will the folding of the membrane protein be easily affected ? and it will be expressed in bacteria or eukaryotic ??
we already got fusion protein of this same membrane protein with gfp and it works fine. We will express this protein in eukaryotic suspension cell culture.......