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Publication Number
US-11326176-B2
Publication Date
2022-05-10
Expiration Date
Abstract
A method of in vivo assembly of a recombinant micelle including: introducing a plasmid into a plant cell, wherein: the plasmid includes a segment of deoxyribonucleic acid (DNA) for encoding a ribonucleic acid (RNA) for a protein in a casein micelle, the segment of DNA is transcribed and translated; forming recombinant casein proteins in the plant cell, wherein: the recombinant casein proteins include a κ-casein and at least one of an αS1-casein, an αS2-casein, a β-casein; and assembling in vivo a recombinant micelle within the plant cell, wherein: an outer layer of the recombinant micelle is enriched with the κ-casein, an inner matrix of the recombinant micelle include at least one of the αS1-casein, the αS2-casein, the β-casein.
Core Innovation
A plant-cell system is used for in vivo assembly of a recombinant micelle by introducing a plasmid into a plant cell. The plasmid comprises DNA encoding RNA for at least two ruminant casein proteins, and the DNA is transcribed and translated in the plant cell, forming recombinant casein proteins in the plant cell. The recombinant casein proteins comprise a κ-casein and at least one of an αS1-casein, an αS2-casein, and a β-casein.
In vivo assembly of the recombinant micelle occurs within the plant cell so that an outer layer of the recombinant micelle is enriched with the κ-casein. An inner matrix of the recombinant micelle comprises at least one of the αS1-casein, the αS2-casein, and the β-casein. The system includes targeting via N-terminal signal peptides and post-translational processing of the recombinant casein proteins as part of forming recombinant micelles.
The plant-cell plasmid system can further include promoter and N-terminal signal peptide DNA, and can include genetic elements such as selectable and/or screenable markers. The system can also include interference RNA to suppress expression of native seed storage proteins, as well as transcription units intended to modulate intracellular mineral pools. Example embodiments are described for plant species including Arabidopsis and soybean, with characterization of in vivo micelles by protein composition and size-range measurements.
Claims Coverage
The patent includes two independent claims that cover in vivo assembly of a recombinant micelle in a plant cell and a plasmid configured for such in vivo assembly. Across the independent claims, the core coverage centers on κ-casein together with αS1-, αS2-, or β-caseins, DNA transcription and translation in the plant cell, and assembly in vivo such that κ-casein is enriched in the outer layer while αS1-, αS2-, or β-caseins comprise the inner matrix.
In vivo assembly of a recombinant micelle within a plant cell by plasmid transcribed and translated DNA
A method of in vivo assembly of a recombinant micelle comprising introducing a plasmid into a plant cell, wherein the plasmid comprises DNA encoding RNA for at least two ruminant casein proteins, wherein the DNA is transcribed and translated in the plant cell, forming recombinant casein proteins in the plant cell, and assembling in vivo a recombinant micelle within the plant cell.
Outer layer enriched with κ-casein and inner matrix comprising at least one of αS1-, αS2-, and β-caseins
Assembling in vivo a recombinant micelle within the plant cell wherein an outer layer of the recombinant micelle is enriched with the κ-casein and an inner matrix of the recombinant micelle comprises at least one of the αS1-casein, the αS2-casein, and the β-casein.
A plasmid for in vivo assembly of a recombinant casein micelle with promoter and N-terminal signal peptide
A plasmid for the in vivo assembly of a recombinant casein micelle comprising DNA encoding at least two ruminant casein proteins comprising a κ-casein and at least one of an αS1-casein, an αS2-casein, a β-casein, and DNA encoding a promoter and an N-terminal signal peptide.
At least two ruminant casein proteins including κ-casein and at least one of αS1-, αS2-, or β-casein for micelle assembly
DNA encoding at least two ruminant casein proteins comprising a κ-casein and at least one of an αS1-casein, an αS2-casein, and a β-casein, in which the plasmid is configured for in vivo assembly of a recombinant casein micelle.
Claim coverage emphasizes in vivo plant-cell formation of recombinant casein proteins followed by in vivo micelle assembly characterized by κ-casein enrichment in the outer layer and αS1-, αS2-, or β-caseins in the inner matrix, with an additional independent claim directed to a plasmid encoding the required ruminant casein proteins plus a promoter and an N-terminal signal peptide.
Stated Advantages
Not explicitly described in patent.
Documented Applications
Not explicitly described in patent.
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