gene expresion - (Nov/19/2008 )
Hi,
I want to know about the gene expression.I want to clone two genes in the order from left to right
1. promoter 2. my gene with both start and stop codon. 3.Gus gene of 1Kb 4. terminator.
my dought is transription will proceed from promoter then my gene, will it proceed to the GUS gene or it will stop because of stop codon of my gene then it will continue or not if it stops.,because i have stop codon in my gene,will it makes any problem for transcribing gene and then to terminator i.e kanamycin for selction.
can any help help me in this issue.
Thanking you very much,
Regards,
SVRREDDY.
-ramirddyag-
QUOTE (ramirddyag @ Nov 19 2008, 12:27 PM)
Hi,
I want to know about the gene expression.I want to clone two genes in the order from left to right
1. promoter 2. my gene with both start and stop codon. 3.Gus gene of 1Kb 4. terminator.
my dought is transription will proceed from promoter then my gene, will it proceed to the GUS gene or it will stop because of stop codon of my gene then it will continue or not if it stops.,because i have stop codon in my gene,will it makes any problem for transcribing gene and then to terminator i.e kanamycin for selction.
can any help help me in this issue.
Thanking you very much,
Regards,
SVRREDDY.
I want to know about the gene expression.I want to clone two genes in the order from left to right
1. promoter 2. my gene with both start and stop codon. 3.Gus gene of 1Kb 4. terminator.
my dought is transription will proceed from promoter then my gene, will it proceed to the GUS gene or it will stop because of stop codon of my gene then it will continue or not if it stops.,because i have stop codon in my gene,will it makes any problem for transcribing gene and then to terminator i.e kanamycin for selction.
can any help help me in this issue.
Thanking you very much,
Regards,
SVRREDDY.
Stop codon has nothing to do with transcription. Polyadenylation signal which follows the stop codon determines the transcription termination.
You need another promoter for Gus gene, or insert an IRES cassettee between the two genes; inserted before the polyadenylation signal of your gene.
it will look like either of the following -
promoter-yourgene-polyAsignal-anotherpromoter-Gusgene
promoter-yourgene-IRES-Gusgene-polyAsignal
(yourgene will contain start and stop codons in both cases).
-cellcounter-