Proteolytic inactivation of select proteins in bacterial extracts for improved expression
US10450353B2 · kind B2 · utility
Assignee
Inventors
Key dates
| Filing date | Apr 10, 2017 |
| Grant date | Oct 22, 2019 |
| Priority date | — |
| Expiry date | Apr 10, 2037 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12Y304/21
- WIPO fieldMeasurement
- WIPO sectorInstruments
Abstract
The present disclosure provides modified proteins that are capable of being cleaved by the protease OmpT1. The proteins can be modified in an exposed surface motif to incorporate OmpT1 cleavage sites. Also provided are nucleic acids encoding the modified proteins, bacterial cells that express the modified proteins, and cell free synthesis systems containing modified RF1. The disclosure further provides methods for reducing the deleterious activity of a modified protein in a cell free synthesis system by contacting the modified protein with OmpT1. Also provided are methods for reducing RF1 competition at an amber codon in the cell free synthesis system, and methods for expressing a protein in the cell free synthesis system. The modified proteins of the invention can be used to increase the yield of proteins having non-natural amino acids incorporated at an amber codon.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.