Tuning for high protein production

DNA contains the information to produce all the proteins a cell needs. However, a cell needs regulatory elements to produce the right protein at the right time in sufficient quantity. To preserve energy a cell does not produce more protein than required. However, for biotechnology applications, a cell needs to produce as much of a specific protein as possible to create an efficient production process. In this thesis, the translation regulating elements were studied to then tune them for reliable high protein yields. The 5’UTR was standardized using a bicistronic design. The 3’UTR was found to significantly impact protein production through a yet unknown mechanism, but surprisingly in a very consistent way. Finally, codon usage was studied extensively and we concluded that the first 9 codons mainly dictate the efficiency of translation and not the overall codon usage.  These findings can contribute to more efficient protein production processes in therapeutics, industry and research and might even enable new production processes.