RT Journal Article
SR Electronic
T1 Cell-free translation is more variable than transcription
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 073841
DO 10.1101/073841
A1 Fabio Chizzolini
A1 Michele Forlin
A1 Noël Yeh Martín
A1 Giuliano Berloffa
A1 Dario Cecchi
A1 Sheref S. Mansy
YR 2016
UL http://biorxiv.org/content/early/2016/12/26/073841.abstract
AB Although RNA synthesis can be reliably controlled with different T7 transcriptional promoters during cell-free gene expression with the PURE system, protein synthesis remains largely unaffected. To better control protein levels, a series of ribosome binding sites (RBS) was investigated. While RBS strength did strongly affect protein synthesis, the RBS sequence could explain less than half of the variability of the data. Protein expression was found to depend on other factors besides the strength of the RBS, including GC content. The complexity of protein synthesis in comparison to RNA synthesis was observed by the higher degree of variability associated with protein expression. This variability was also observed in an E. coli cell extract-based system. However, the coefficient of variation was larger with E. coli RNA polymerase than with T7 RNA polymerase, consistent with the increased complexity of E. coli RNA polymerase.