SV40 early promoter read-through - (Jan/15/2007 )
Hello!
I've cut the antibiotic resistance (KanR/NeoR) plus the HSV TK polyA out from pEGFP-N1(using EcoO109I and StuI) and religated the vector. The SV40 early promoter, driving the NeoR, remains in the vector. Is it possible that there is a read-through from this SV40 promoter into my eGFP-reporter gene?
Thanx in advance!
SV40 promoter will not affect the eGFP expression. The promoter is too weak to make any impact when a stronger CMV promoter is driving the expression.
Did u add any other antibiotic resistance gene to ur vector?
Did u add any other antibiotic resistance gene to ur vector?
No i didn't! I'm going to check the effect on gene expression when the antibiotic resistance is deleted from the vector!
I believe that if you removed the Neo poly A signal, that the SV40 promoter could drive expression of GFP independently of CMV. It might be a limited effect based on the expression levels from CMV, but I do believe this could occur.