Expression of biologically active polypeptides in duckweed
AssigneeBiolex Therapeutics, Inc.
InventorsAnne-Marie STOMP, Lynn Dickey, John Gasdaska
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Abstract
Methods, nucleic acid sequences, and transformed duckweed plant or duckweed nodule cultures for the expression and the secretion of biologically active polypeptides from genetically engineered duckweed are provided. Expression of recombinant polypeptides in duckweed is improved by modifying the nucleotide sequence of the expression cassette encoding the polypeptide for improved expression in duckweed. Recovery of biologically active polypeptides from duckweed is improved by linking the biologically active polypeptide to a signal peptide that directs the secretion of the polypeptide into the culture medium.
Claims33
A duckweed plant culture or a duckweed nodule culture, wherein said duckweed plant culture or said duckweed nodule culture is stably transformed with a nucleic acid molecule comprising a coding sequence for α-2b-interferon and an operably linked coding sequence for a signal peptide capable of directing secretion of the α-2b-interferon.
A duckweed plant culture or a duckweed nodule culture, wherein said duckweed plant culture or said duckweed nodule culture is stably transformed with a nucleic acid molecule comprising a nucleotide sequence encoding biologically active α-2b-interferon wherein said nucleotide sequence has been modified for enhanced expression in duckweed.
A nucleic acid molecule comprising the α-2b-interferon-encoding nucleotide sequence set forth in SEQ ID NO:2 operably linked to the signal peptide-encoding nucleotide sequence given in SEQ ID NO:3.
A duckweed plant culture or duckweed nodule culture, stably transformed to express a biologically active recombinant polypeptide from a nucleotide sequence comprising a coding sequence for said recombinant polypeptide and an operably linked coding sequence for a signal peptide that directs secretion of the recombinant polypeptide;
- wherein said signal peptide sequence is selected from the group consisting of:
- (a) the human α-2b-interferon signal peptide;
- (b) the Arabidopsis thaliana chitinase signal peptide;
- (c) the rice α-amylase signal peptide;
- and
- (d) the modified rice α-amylase peptide set forth in SEQ ID NO:7.
A method of producing biologically active recombinant polypeptide in a duckweed plant culture or a duckweed nodule culture, comprising the steps of:
- (a) culturing within a duckweed culture medium a duckweed plant culture or a duckweed nodule culture, wherein said duckweed plant culture or said duckweed nodule culture is stably transformed to express said biologically active recombinant polypeptide, and wherein said biologically active recombinant polypeptide is expressed from a nucleotide sequence comprising a coding sequence for said recombinant polypeptide and an operably linked coding sequence for a signal peptide that directs secretion of the recombinant polypeptide;
- and
- (b) collecting said recombinant polypeptide from the duckweed culture medium;
- wherein said signal peptide is selected from the group consisting of:
- (i) the human α-2b-interferon signal peptide;
- (ii) the Arabidopsis thaliana chitinase signal peptide;
- (iii) the rice α-amylase signal peptide;
- and
- (iv) the modified rice α-amylase peptide set forth in SEQ ID NO:7.
Dependencies are inferred from the claim text.