I have already used some strategies such as codon optimization, Kozak sequence and ER signal for enhancing transgene expression in plant host. Is there any other strategy for enhancing gene expression level in transgenic plants?
1. You can use a strong promoter
2. Some 'introns' can increase gene expression (see attached papers); also see 'Intron-mediated enhancement' ( http://en.wikipedia.org/wiki/Intron-mediated_enhancement )
3. If affordable, you can also direct your transgene to a proven locus with high expression characteristic using current molecular tool such as sequence-specific nucleases (ex. TALEN, CRISPR/Cas,....).
Google for ribosomal ramp. Choose the first 10 codons wisely - nott too rare but also not too good ones. Low GC content in the leader and first amino acids may also help the ribosomes to get started
"28 nt long synthetic 50UTR (synJ) as an enhancer of transgene expression in
dicotyledonous plants" - a paper that may be interesting.
Or the paper of the "super strong plant promoter" http://www.ncbi.nlm.nih.gov/pubmed/14756302
Mind, not all codon optimization tools are good. Many will only pick frequently used codons.
Dear Maziar
I think the best way is using a MAR (matrix attachment region) in transgene cassette. Nuclear matrix attachment regions (MARs) are genomic DNA sequences, located at the physical boundaries of chromatin loops. MARs may influence the structure of transgene chromatin and/or its ability to interact with other sequences in the nucleus.Inclusion of MARs in transgene cassettes enhances their expression and reduces position-effect variations in the transgenic host.
good luck
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