Rumor also has it that more than one "bad" codon in a row, particular
near the beginning, can be extra-bad. For one small, A/T rich
protein, we simply "fixed" a pair of bad Arg codons near the
N-terminus at the same time as recloning it, by using the N-terminal
cloning primer to do the mutagenesis. Combined with using codon-plus
E coli, expression improved dramatically.
Good luck!
Phoebe Rice
At 09:29 AM 2/1/2008, Looney, Andrea Lynn (Andrea Hevrdeys) wrote:
Dear All,
I am getting very little (not zero) expression of my protein. I am
curious to know if it is worthwhile to codon optimize my gene, which
is ~1200bp, for E.coli. Can you have "too much of a good
thing"? Can codon optimizing overwhelm the cell or in some way
cause death or lowered expression?
Thank you all,
Andrea L. Hevrdeys
---------------------------------------------------------------------------------------------------------------------------
Phoebe A. Rice
Assoc. Prof., Dept. of Biochemistry & Molecular Biology
The University of Chicago
phone 773 834 1723
fax 773 702 0439
http://bmb.bsd.uchicago.edu/Faculty_and_Research/01_Faculty/01_Faculty_Alphabetically.php?faculty_id=123
http://www.nasa.gov/mission_pages/cassini/multimedia/pia06064.html
