Method for generating split, non-transferable genes that are able to express an active protein product
AssigneeNew England Biolabs, Inc.
InventorsMing-Qun Xu, Thomas C. Evans, Sriharsa Pradhan, Donald G. Comb, Henry Paulus, Luo Sun, Lixin Chen, Inca Ghosh
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Abstract
A new type of transgene system is disclosed which allows efficient protein expression in a target host such as a plant, but avoids the undesirable result of the migration of the transgene into related host system and/or to the environment via the pollen. The methods described herein may also be applied to the expression of virtually any protein of interest (e.g. a toxic protein) in eukaryotic (yeast, insect, mammalian cells, etc.) and prokaryotic (E. coli, etc.) organisms.
Claims18
A method of reconstituting a target protein from protein fragments in a plant, comprising:
- (a) splitting a gene encoding a target protein into at least two DNA fragments;
- (b) separating the DNA fragments of step (a) to prevent transmission of the gene to other plants;
- wherein one of the DNA fragments coding for a portion of the target protein is compartmentalized in the nucleus, and the other DNA fragment coding for another portion of the target protein is compartmentalized in the chloroplast;
- (c) expressing the DNA fragments of step (b) within the plant to produce the corresponding fragments of the target protein;
- and
- (d) reconstituting the target protein from the protein fragments in the plant.
A method of preventing transmission to a second plant of a gene coding for a target protein in a first plant, comprising:
- (a) splitting the gene encoding the target protein into at least two DNA fragments;
- and
- (b) separating the DNA fragments of step (a) wherein one of the DNA fragments coding for a portion of the target protein is compartmentalized in the nucleus of a host cell in the first plant, and the other DNA fragment coding for another portion of the target protein is compartmentalized in the chloroplasts of the host cell;
- and
- (c) preventing transmission of the gene coding for the target protein to the second plant.
Dependencies are inferred from the claim text.