I recently saw the paper:
"The +4G Site in Kozak Consensus Is Not Related to the Efficiency of Translation Initiation"
Article The +4G Site in Kozak Consensus Is Not Related to the Effici...
Which uses Bioinformatics to argue that the conserved +4 G in the Kozak sequence (below) is because the second amino acid is typically alanine. And that encoding alanine is important, not +4 G per se.
(A/G)CCATGG
Some of my plasmids have n-terminal affinity tags and the second codon is GAC, which encodes Asp, not Ala. This GAC codon satisfies the +4 G of the consensus Kozak sequence, but it does not encode Alanine like the paper suggests.
Does anyone know how alanine as the second amino acid affects expression in mammalian systems? Should I change my plasmids to alanine for the second codon? I couldn't find any papers on this.
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