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Selected Publication:

Mairhofer, J; Wittwer, A; Cserjan-Puschmann, M; Striedner, G; .
(2015): Preventing T7 RNA Polymerase Read-through Transcription-A Synthetic Termination Signal Capable of Improving Bioprocess Stability.
ACS Synth Biol. 2015; 4(3):265-273 FullText FullText_BOKU

Abstract:
The phage-derived T7 RNA polymerase is the most prominent orthogonal transcriptions system used in the field of synthetic biology. However, gene expression driven by T7 RNA polymerase is prone to read-through transcription 11 due to contextuality of the T7 terminator.. The native T7 emulator has a termination efficiency of approximately 80% and therefore provides insufficient insulation of the expression unit. By using a combination of a synthetic T7 termination signal with two well-known transcriptional terminators (rrnBT1 and T7), we have been able to increase the termination efficiency to 99%. To characterize putative effects of an enhanced termination signal on product yield and I process stability, industrial-relevant fed batch cultivations have been performed. Fermentation of a E. coli HMS174(DE3) strain carrying a pET30a derivative containing the improved termination signal showed a significant decrease of plasmid copy number (PCN) and an increase in total protein yield under standard conditions.
Authors BOKU Wien:
Cserjan Monika
Mairhofer J├╝rgen
Striedner Gerald

Find related publications in this database (Keywords)
recombinant protein expression
plasmid vector design
T7 RNA polymerase
con textuality
T7 terminator
read-through transcription
insulator


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