Posted 22 January 2013 - 01:02 PM
I am wondering if anybody has the same experience as me.
I generated lentivirus with a MSCV vector with GFP at the N terminal, encoding full length of my protein.
With the viral stocks, I infected multiple cancer cell lines. i could see green fluorescence under microscope and sorted the cells by flow cytometry. I did see lots of fluorescence after sorting. But the expression levels of the protein didn't change at all, tested by western blot.
I don't understand. If it is because of the cell toxicity of my protein.....? But why i could see so much fluorescence?
I am considering to switch another vector. But i still don't understand what happened to my protein. How could the GFP and my protein be separated?
If anybody can help me figure this out?
Posted 26 January 2013 - 08:27 PM
Posted 27 January 2013 - 12:45 PM
In the vector, my gene is actually in front of GFP, another word, GFP is at the N-terminal of my gene.