I want to express a human gene in the tobacco plant. My gene does not have any kozak consensus sequence or even kozak like sequence upstream of the ATG.
Do I need to insert it in my construction or not?
What is your suggestion for Kozak sequence? CCACC before [ATG] or
GCCACC then [ATG] or CGCCACC [ATG] (G?)
How come the downstream G after ATG can be added/altered without changing the CDS of the protein?
I do not think that Kozak consensus sequence applies to plants, but some equivalent sequences have been reported: http://www.ias.ac.in/jgenet/Vol78No2/jg440.pdf
Based on observations with numerous plant genes, I would suggest CCAAC[ATG]. The downstream G would change the amino acid after the met, but plants don't seem to absolutely require a G after the start codon. Do make sure that the promoter and 5' leader do not contain any other ATGs followed by Stop codons, as this will reduce translation of your gene. I have a paper that refers to this you might use to obtain more relevant citations [Bassett et al. "Multiple transcripts of a gene....." MGG 271:752-760 (2004). You should also be aware that plants will sometime "process" animal mRNAs, so check your CDS for GT...adenine/thymine rich sequence of varying lengths...AG. The GT and AG bases are the start and end of the intron recognition. If you have AT rich seqs between these, they might be removed by the plants. Good luck!
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