Plasmid addiction system to drive desired gene expression
AssigneeSumitomo Chemical Company, Limited
InventorsMatthew de la Pena Mattozzi, Daniel Kim, Sonya Clarkson
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Abstract
The present invention relates to a Plasmid Addiction System for the stabilization of expression plasmids encoding proteins of interest. The invention uses a succinate cycle optimization to ensure the expression of plasmid(s) of interest. By ensuring that plasmids of interest contain genes necessary in the succinate cycle, the system ensures that the passage of the plasmid to daughters and therefore improves the efficiency of production and expression of genes and/or products of interest.
Claims10
A process for producing a recombinant protein of interest in a transformed bacterial host, comprising:
- a) introducing one or more recombinant plasmids into said bacterial host cell, wherein said one or more recombinant plasmids comprise a cloned DNA sequence comprising all or part of at least two genes each encoding a functional essential succinate pathway enzyme that is integral to the survival of the host cell and the one or more recombinant plasmids also encode at least one gene of interest coding for a protein product, and wherein the one or more recombinant plasmids are devoid of an antibiotic resistance gene;
- b) selecting surviving colonies of the transformed host containing the one or more recombinant plasmids;
- and,
- c) using the surviving colonies for fermenting bacterial colonies to allow the expression of said protein product;
- and
- wherein the at least two genes are sucAB and sucCD, and
- wherein the bacterial host is a mutant with eliminated activity of sucAD or sucABCD.
Dependencies are inferred from the claim text.