| Products/Services Used | Details | Operation |
|---|---|---|
| Gene Synthesis | All cistrons contained a single gene except the two-subunit proteins GlyRS and PheRS, which were maintained as two-gene cistrons as found in wild type E. coli. Each gene (flanked by a VSV terminator) was synthesized from oligodeoxyribonucleotides, cloned and sequence-verified by GenScript (NJ, USA; total cost was $36492 in 2008), then subcloned into a BioBrick vector afterastandardT7promoter/lacoperator/RBS(Figure1B). See Supplementary Methods for more details. | Get A Quote |
Two of the many goals of synthetic biology are synthesizing large biochemical systems and simplifying their assembly. While several genes have been assembled together by modular idempotent cloning， it is unclear if such simplified strategies scale to very large constructs for expression and purification of whole pathways. Here we synthesize from oligodeoxyribonucleotides a completely de-novo-designed， 58-kb multigene DNA. This BioBrick plasmid insert encodes 30 of the 31 translation factors of the PURE translation system， each His-tagged and in separate transcription cistrons. Dividing the insert between three high-copy expression plasmids enables the bulk purification of the aminoacyl-tRNA synthetases an... More
