Question about GFP
Posted 12 February 2012 - 11:43 PM
Posted 13 February 2012 - 12:15 AM
Mybe your interested proteins have some toxicity or negetive effect on the cell growth, so they cannot compete with the cells without GFP and your interested genes.
If you have the strict and logical control, your results are still available and useful and can be published in journals.
Posted 27 March 2012 - 03:22 AM
Posted 29 March 2012 - 04:40 PM
Posted 20 November 2014 - 10:27 PM
It happens to my GFP stable cells too. I construct a LNCaP/EGFP stable cell lines by cloning pickup. Everything goes well. The cells have very good green fluorescence first. but the fluorescence of my cells suddenly become weaker and weaker. I thought this clone might be unstable clone, so I picked another 3, but all have the similar result. I almost gave up, but I find that the GFP expression of the cell can be recovered by increasing the G418 concentration. When I increase the concentration to 1000 ug/ml and culture them for around 1 week, all the cells show very very strong fluorescence again and cells are also healthy. The only problem is the cell growth rate, very slow. BTW, I use the high pure G418. So I think the GFP gene is not kicked out, might be in hybernation. You can wake it up by G418.