Expression of toxic genes in Methylorubrum extorquens with a tightly repressed, cumate-inducible promoter

  • Methylorubrum extorquens is an important model methylotroph and has enormous potential for the development of C1-based microbial cell factories. During strain construction, regulated promoters with a low background expression level are important genetic tools for expression of potentially toxic genes. Here we present an accordingly optimised promoter, which can be used for that purpose. During construction and testing of terpene production strains harbouring a recombinant mevalonate pathway, strong growth defects were observed which made strain development impossible. After isolation and characterisation of suppressor mutants, we discovered a variant of the cumate-inducible promoter PQ2148 used in this approach. Deletion of 28 nucleotides resulted in an extremely low background expression level, but also reduced the maximal expression strength to about 30% of the original promoter. This tightly repressed promoter version is a powerful module for controlled expression of potentially toxic genes in M. extorquens.

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Author:Laura PöschelORCiDGND, Elisabeth Gehr, Paulina Jordan, Frank Sonntag, Markus BuchhauptORCiDGND
Parent Title (English):Antonie van Leeuwenhoek
Publisher:Springer Science + Business Media B.V
Place of publication:Dordrecht [u.a.]
Document Type:Article
Date of Publication (online):2023/09/26
Date of first Publication:2023/09/26
Publishing Institution:Universitätsbibliothek Johann Christian Senckenberg
Release Date:2023/11/13
Tag:Alphaproteobacteria; Background expression; Cumate; Inducible Promoter; Methylorubrum extorquens AM1
Page Number:10
Dewey Decimal Classification:5 Naturwissenschaften und Mathematik / 57 Biowissenschaften; Biologie / 570 Biowissenschaften; Biologie
Licence (German):License LogoCreative Commons - CC BY - Namensnennung 4.0 International