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US 10,155,951 B2Granted
Methods and systems for engineering bacterial systems to shield toxicity during non-native protein expression and purification
AssigneeLawrence Livermore National Security, LLC
InventorsMimi Cho Yung, Feliza A. Bourguet, Timothy S. Carpenter, Matthew A. Coleman
Life of this patent
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Abstract
The invention described herein relates to methods, systems, compositions and cells to provide one or more toxic non-native proteins in a cell, wherein the one or more toxic non-native proteins are contained in at least one empty microcompartment within the cell.
Claims27
A method to provide a toxic non-native protein in a cell, the method comprising
- introducing into the cell at least one first polynucleotide encoding at least one microcompartment protein, the at least one polynucleotide operatively linked to one or more first regulatory elements leading to the expression of the at least one microcompartment protein in the cell, the at least one microcompartment protein capable of assembling with one or more same and/or different microcompartment proteins to form at least one empty microcompartment within the cell;
- and
- introducing into the cell at least one second polynucleotide encoding for one or more toxic non-native proteins capable of reacting with a native membrane substrate with a reaction resulting in a cell damage, each of the one or more toxic non-native proteins operably linked to a leader peptide capable of directing expressed proteins to the at least one empty microcompartment, the at least one second polynucleotide operably linked to one or more second regulatory elements leading to the expression of the at least one toxic non-native protein operably linked to the leader peptide in the cell;
- to obtain the toxic non-native protein within the at least one empty microcompartment within the cell.
1 more element
A system for shielding a bacterial cell from toxicity during intracellular production of a toxic non-native protein, the system comprising
- at least one first polynucleotide encoding at least one microcompartment protein operatively linked to one or more first regulatory elements leading to the expression of the at least one microcompartment protein in the cell, the at least one microcompartment protein capable of assembling with one or more same and/or different microcompartment proteins to form at least one empty microcompartment within the cell;
- and
- at least one second polynucleotide encoding for at least one toxic non-native protein capable of reacting with a native membrane substrate with a reaction resulting in a cell damage each of the one or more toxic non-native proteins operably linked to a leader peptide capable of directing expressed proteins to the at least one empty microcompartment, the at least one second polynucleotide operatively linked to one or more second regulatory elements leading to the expression of the at least one toxic non-native protein operably linked to the leader peptide.
Dependencies are inferred from the claim text.