Inhibition of peptide cleavage in plants
AssigneeLarge Scale Biology Corporation
InventorsKathleen M. Hanley, Fakhrieh S. Vojdani, Long V. Nguyen, Wayne P. Fitzmaurice
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Abstract
The invention provides a system for expressing a foreign peptide in a plant cell, wherein the foreign protein is sensitive to a protease activity in the plant cell, by introducing into a plant cell a polynucleotide, which encodes the foreign protein and an another polynucleotide, which encodes a genetic element capable of reducing the protease activity in the plant cell. The invention also provides for plant cells, which incorporate this system, and for methods of reducing the proteolysis of the foreign protein expressed in a plant cell by using this system.
Claims37
A host cell comprising one or more polynucleotides, wherein said one or more polynucleotides encode a protein of interest and a genetic element capable of reducing a protease activity in a host cell or fluids, wherein said one or more polynucleotides is capable of expressing said protein of interest in the host cell, wherein said protease activity is capable of cleaving said protein of interest, wherein said protein of interest is non-native to the host cell.
25 The plant cell comprising the plant cell according to claim 6.
A polynucleotide comprising a substantially similar or complementary sequence of at least a part of the coding sequence, or one or more fragments, of a Nicotianalisin protease which is not identical with another protease.
A method of reducing the amount of a protein of interest cleaved by a hydrolase activity in a host cell, comprising the steps of:
- (a) introducing a polynucleotide into a host cell, wherein said polynucleotide comprises a genetic element capable of reducing a hydrolase activity in said host cell;
- and
- (b) expressing a protein of interest in said host cell, wherein said protein of interest is capable of expression in said host cell, wherein said protein of interest is capable of being cleaved by said hydrolase activity;
- whereby the amount of protein of interest cleaved by said hydrolase activity in said host cell is reduced compared to the amount of protein of interest cleaved by said hydrolase activity in another host cell in which said polynucleotide is not introduced.
A vector containing a genetic element capable of reducing protease activity and a polynucleotide encoding a protein of interest.
A composition of purified Nicotianalisin having a specific activity greater than 100 units/mg protein.
Dependencies are inferred from the claim text.