primer design - over expression (Jul/03/2008 )
Hi all
i designed a primer to change translation initiation region of some protein cDNA and i studid the level of expression of it in E. coli. i changed just 11 codons in Nterminal without changing amino acid sequences. some one asked me : why you changed just 11 codons ?? and i'm thinking about it ... so any one who can help????
-mona_bola-
QUOTE (mona_bola @ Jul 3 2008, 06:48 PM)
Hi all
i designed a primer to change translation initiation region of some protein cDNA and i studid the level of expression of it in E. coli. i changed just 11 codons in Nterminal without changing amino acid sequences. some one asked me : why you changed just 11 codons ?? and i'm thinking about it ... so any one who can help????
i designed a primer to change translation initiation region of some protein cDNA and i studid the level of expression of it in E. coli. i changed just 11 codons in Nterminal without changing amino acid sequences. some one asked me : why you changed just 11 codons ?? and i'm thinking about it ... so any one who can help????
how do you know only 11 matter ?
why not 10 or 3 ?
if you have references, then go for it, but if you only chose this number, it's indeed questionable.
... and by "changing the codons" do you mean introducing silent base changes ? I'm not sure I understood your problem correctly... Grosso modo, you wanna see what is the determining sequence before the translation start and how mutating them affects translation ?
-ph3no-