Non-glycosylated transferrin expressed in monocots
AssigneeVentria Bioscience
InventorsDeshui Zhang, Ning Huang, Diane Phuong Nguyen, Paula Bryan
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Abstract
Disclosed are compositions and methods of making non-glycosylated transferrin protein in transgenic monocot plants.
Claims18
A method of producing a recombinant, non-glycosylated transferrin protein in rice plant seeds, said method comprising the steps of:
- (a) transforming a rice plant cell with a chimeric gene comprising
- (i) a promoter from a gene of a seed maturation-specific monocot plant storage protein;
- (ii) a first DNA sequence, operably linked to said promoter, the first DNA sequence encoding a signal sequence targeting a polypeptide linked thereto to a protein storage body of a monocot plant seed cell;
- and
- (iii) a second DNA sequence, linked in translation frame with the first DNA sequence, encoding a mammalian transferrin protein, wherein the first DNA sequence and the second DNA sequence together encode a fusion protein comprising the signal sequence and the transferrin protein;
- (b) growing a rice plant from the transformed rice plant cell for a time sufficient to produce seeds containing the transferrin protein;
- and
- (c) harvesting the seeds from the plant.
A seed-derived composition comprising a non-glycosylated transferrin protein, and at least one pharmaceutically acceptable excipient or nutrient, wherein the non-glycosylated transferrin protein is produced in a rice plant containing a nucleic acid sequence encoding the transferrin protein and is extracted from seed harvested from the rice plant.
A method for making a cell culture medium, said method comprising the steps of:
- (a) obtaining a rice plant stably transformed with a chimeric gene having construct comprising
- (i) a seed maturation-specific promoter;
- (ii) a leader DNA sequence encoding a transit sequence targeting a linked polypeptide to a protein storage body of a rice plant seed cell;
- wherein said leader DNA sequence is operably linked to said promoter;
- (iii) a protein-coding sequence encoding a transferrin protein;
- (b) cultivating the transformed plant under seed-maturation conditions;
- (c) harvesting the seeds from the cultivated plant;
- (d) extracting the harvested seeds with an aqueous solution, thereby obtaining an extract of water soluble plant components comprising at least 3% by total protein weight of non-glycosylated transferrin protein;
- (e) purifying the transferrin protein from the aqueous solution;
- and
- (f) adding the purified transferrin protein to a cell culture medium.
A serum-free cell culture medium comprising an extract of rice seed comprising seed-expressed non-glycosylated transferrin protein.
Dependencies are inferred from the claim text.