Large vectors and methods for high-yield production
AssigneeKatholieke Universiteit Leuven
InventorsMelody Janssen, Joeri Auwerx, Kai Dallmeier, Nicolas Ongenae, Cédric Vansalen, Nesya Goris
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AssigneeKatholieke Universiteit Leuven
InventorsMelody Janssen, Joeri Auwerx, Kai Dallmeier, Nicolas Ongenae, Cédric Vansalen, Nesya Goris
Estimating the term…
Provided herein are methods for the production of a vector with a size of at least 16 kb from bacterial cells comprising the consecutive steps of a) obtaining bacterial cells comprising a vector with a size of at least 16 kb, comprising an inducible origin of replication, b) inoculating culture medium with the bacterial cells comprising the vector, c) culturing the bacterial cells in the culture medium, d) adding one or more inducers of said inducible origin of replication to the culture medium when the bacterial culture has reached an optical density at 600 nm (OD600) of at least 20, e) further culturing the bacterial cells in the culture medium, f) optionally separating the bacterial cells from the culture medium, and g) recovering the plasmid from the bacterial cells. Also provided herein are vectors with a size of at least 16 kb comprising an inducible origin of replication for use in such methods.
A vector with a size of at least 16 kb comprising an inducible origin of replication, wherein the inducible origin of replication comprises a nucleic acid sequence having at least 95% sequence identity with the nucleic sequence as set forth in SEQ ID NO 9.
A method for the production of a vector with a size of at least 16 kb from bacterial cells, wherein the vector comprises an inducible origin of replication that comprises a nucleic acid sequence having at least 95% sequence identity with the nucleic sequence as set forth in SEQ ID NO:
An inducible origin of replication comprising a nucleic acid sequence having at least 95% sequence identity with the nucleic sequence as set forth in SEQ ID NO:
Dependencies are inferred from the claim text.