Method for producing phosphoserine incorporated proteins by using SepRS mutants and EF-Tu mutants
US9322044B2 · kind B2 · utility
Assignee
Inventors
Key dates
| Filing date | Mar 14, 2014 |
| Grant date | Apr 26, 2016 |
| Priority date | — |
| Expiry date | Apr 22, 2034 |
Classification
- Technology area (CPC Y)Emerging Cross-Sectional Technologies
- CPC primaryY02P20/52
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
The present invention relates to a method of producing a phosphorylated protein using a SepRS (O-phosphoseryl-tRNA synthetase) mutant and an EF-Tu mutant, which have increased activity. More specifically, the invention relates to a method of producing a phosphorylated protein by incorporating phosphoserine into the specific position of a target protein or polypeptide using tRNASep serving to recognize at least one codon in the mRNA of the target protein or polypeptide, an O-phosphoseryl-tRNA synthetase (SepRS) mutant selected by a molecular evolution technique and serving to aminoacylate tRNASep with phosphoserine (Sep), and an EF-Tu mutant serving to bind and deliver Sep-tRNASep to the ribosome.According to the invention, a phosphorylated protein can be produced in an amount of mg per liter using the SepRS and EF-Tu mutants. Thus, the invention is useful for the production of various phosphorylated proteins, including phosphorylated enzymes, the production of antibodies, the fabrication of protein chips, and cell-based screening for new drug development.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.