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Hi Reza, We have also used E. coli-optimized constructs in insect cell systems with no problems. It might worth checking a few different insect cell lines. We have a case when little expression is observed in Sf9 and Sf21, but massive amount (50 mg/L secreted) is obtained from High Five (same as T. ni from other companies). I have heard that screening a few different media is a good idea, but I have not experience this problem first hand. PCR the virus directly to see if the expression cassette is still there is probably also a good idea. Best, Pun On Tue, Feb 16, 2016 at 8:47 PM, Reza Khayat <[log in to unmask]> wrote: > Hi, > > > We have an E. coli codon optimized gene that we would like to try > expressing in insect cells. The protein is not expressing and we are > concerned that it may be because of the codon optimization. Could this be a > possibility, or are the insect optimized codons similar to that of E. coli? > Thanks. > > > Best wishes, > Reza > > > Reza Khayat, PhD > Assistant Professor > City College of New York > Department of Chemistry > New York, NY 10031 >