It depends. It is for basic science lentiviruses are very good on proliferating and non-proliferating cells, however it can be harsh for some cells. Yet the transaction efficiency can be very high 80-90%. Retroviruses would only transduce dividing cells and are also very effective and can be gentler depending on the cell type. For highly proliferating cells I personally would use retroviruses.

If it is to create therapeutic models, adenoviruses are good because they do not implant themselves into the genome; however, the expression decreases over time.

Transfections, I would only use them if viruses are out of reach or if the cells used get very high transfection efficiencies (like some cell lines)

Thank you for your answer, Alejandro. I really need, first, choose the plasmid vectors which contain the detection system. Do you know about it?

I used pEGFPN-1 (GFP) from Clontech, it worked well for me. I am not sure if they still have them, please check with them. What is the size of your protein that you would like to tag?

Hi, Sohail. Thanks for your help. My protein is about 37 kDa.

In which bacterium did you use this plasmid?

The kit comes with XL1-Blue supercompetent cells, they work very well. You can use any other competent cell, but the transformation efficient would not be same with XL1 blue. DHa is also a good host strain.

Looking at the size of your protein, tagging with GFP may not be a good idea. GFP is approximately 25-26 kDa and you might get into situation of "protein misfolding or structure/function problems" and it would not be a good strategy for protein localization study. However, I am not sure about your goal here. If you still want to go ahead with GFP tagging, please remove stop codon from vector when you do mutagenesis. Otherwise, you won't get GFP signal. If possible please explain, the goal of your expt.

Best wishes

SSQ