Thanks for your reply Dr. Ligterink, pUAST kind of vector with GFP is already exist i.e. UAS-GFP, but we want to clone our desire gene in that one. actually i exactely want to do translational fusion of my gene and GFP because i want to check the effect of the that peptide having few amino acid in it.

Dr. Aaron, You mean to say that we have to place GFP towards N and C terminal of our desire gene. anmd how I check N terminal and C terminal fusion,

Actually sir i m sorry if it is too basic question but i don't have bioinformatic knowledge

Actually Sir, I want to clone my desire gene and GFP togather, so is it possible if yes then what should i do, for this should we do ordered for the synthetic peptide from any company,having both our desired sequence and GFP.

Respected Dr. Wilco,

I want to check the express pattern of this complete sequence with my desire gene and GFP in drosophila.

So Sir please explain me in detail from staring what should i do? actually i don't have much knowledge in this, i m in my first year of phd. So please tell me what bioinformatics software we do need to use, what other things are necessary for this experiment.

I will be very thankful to you.

Yes sir, exactly i want to check the localization and movement of our desire protein in living cell.

So for this what should i do?

I know i m irritating you so much? but Sir i will be very thankful to you for helping me.

We usually add a linker between desire protein and GFP, The linker sequence is (4G1S)*2 ,which can avoid misfolding of eitheir protein