In vivo detection of proteins interaction based on adenylate cyclase hybrid system
US10760072B2 · kind B2 · utility
Assignee
Inventors
Key dates
| Filing date | Jul 15, 2015 |
| Grant date | Sep 1, 2020 |
| Priority date | — |
| Expiry date | Jun 2, 2037 |
Classification
- Technology area (CPC G)Physics
- CPC primaryG01N2333/988
- WIPO fieldMeasurement
- WIPO sectorInstruments
Abstract
The present invention relates to a method to detect the interaction between a target ligand and a moiety of interest using an adenylate cyclase enzyme (AC) and calmodulin (CaM) as interacting partners, said method comprising:i) expressing in a suitable host cell:(a) a low number of molecules of a first chimeric polypeptide containing AC, and(b) a low number of molecules of a second chimeric polypeptide containing CaM,wherein said AC in said first chimeric polypeptide and/or said CaM in said second chimeric polypeptide has decreased affinity for its interacting partner,wherein said AC in said first chimeric polypeptide is fused to a moiety of interest and said CaM in said second chimeric polypeptide is fused to a target ligand, or conversely,and wherein, when said moiety of interest and said target ligand interact, said AC is activated, andii) detecting the activation of said AC.The present inventors herein show that only one AC/CaM complex per cell is sufficient to confer a selectable trait to the host cell. Unexpectedly, even less than one AC/CaM complex per cell can be sufficient to confer a selectable trait to the host cell. This surprising result confers a very high sensitivity…
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.