Patent ID: 8853179

Claim:
A method of improving full-length protein expression efficiency comprising: a) providing a polynucleotide comprising: i) a coding sequence for the full-length protein; ii) a primary initiation codon that is upstream of the coding sequence of the full-length protein, said primary initiation codon encoding the first amino acid of the coding sequence of the full-length protein; and iii) one or more secondary initiation codons located within the coding sequence of the full-length protein downstream of the primary initiation codon; and b) mutating the one or more secondary initiation codons located within the coding sequence of the full-length protein downstream of the primary initiation codon, wherein the mutation results in a decrease in initiation of protein synthesis at the one or more secondary initiation codons, thereby increasing expression efficiency of the full-length protein initiated at the primary initiation codon, wherein mutating the one or more secondary initiation codons located within the coding sequence of the full-length protein downstream of the primary initiation codon comprises mutating one or more nucleotides such that the amino acid sequence of the protein remains unaltered.