Recombinant carrier molecule for expression, delivery and purification of target polypeptides
First Claim
1. A carrier protein comprising a recombinant carrier molecule, wherein the recombinant carrier molecule has at least 90% amino acid identity to an amino acid sequence of a modified lichenase B (licB) polypeptide that is based on a rearranged amino acid sequence of the Clostridium thermocellum wild-type licB (wild-type licB) amino acid sequence encoded by the nucleic acid sequence set forth in SEQ ID NO:
- 18, wherein the modified licB polypeptidecomprises original N-terminal and C-terminal regions that correspond in sequence to N- and C-terminal polypeptides obtained by splitting the amino acid sequence encoded by SEQ ID NO;
18 within the loop region defined by amino acid residues 82-94 of the amino acid sequence encoded by SEQ ID NO;
18,wherein the modified licB polypeptide comprises the original C-terminal region fused upstream of the original N-terminal region, andwherein the recombinant carrier molecule optionally has a heterologous polypeptide comprising a therapeutic polypeptide or a disease-associated epitope fused thereto.
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Accused Products
Abstract
Recombinant carrier molecules having amino acid sequences from thermostable enzymes and methods of use for expression, recovery and delivery of foreign sequences (peptides and polypeptides) produced in different systems (bacteria, yeast, DNA, cell cultures such as mammalian, plant, insect cell cultures, protoplast and whole plants in vitro or in vivo are provided. The recombinant carrier molecule using sequences from lichenase B (Lic B) were also made and used as part of carrier protein to express, recover and deliver a variety of target polypeptides of interest.
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Citations
13 Claims
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1. A carrier protein comprising a recombinant carrier molecule, wherein the recombinant carrier molecule has at least 90% amino acid identity to an amino acid sequence of a modified lichenase B (licB) polypeptide that is based on a rearranged amino acid sequence of the Clostridium thermocellum wild-type licB (wild-type licB) amino acid sequence encoded by the nucleic acid sequence set forth in SEQ ID NO:
- 18, wherein the modified licB polypeptide
comprises original N-terminal and C-terminal regions that correspond in sequence to N- and C-terminal polypeptides obtained by splitting the amino acid sequence encoded by SEQ ID NO;
18 within the loop region defined by amino acid residues 82-94 of the amino acid sequence encoded by SEQ ID NO;
18,wherein the modified licB polypeptide comprises the original C-terminal region fused upstream of the original N-terminal region, and wherein the recombinant carrier molecule optionally has a heterologous polypeptide comprising a therapeutic polypeptide or a disease-associated epitope fused thereto. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8)
- 18, wherein the modified licB polypeptide
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9. A method for production of a carrier protein in a plant comprising:
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(a) providing a plant containing an expression cassette having a nucleic acid encoding a carrier protein operably linked to a promoter such that expression of the cassette results in expression of the carrier protein, wherein the carrier protein comprises a recombinant carrier molecule having at least 90% amino acid identity to an amino acid sequence of a modified licB polypeptide that is based on a rearranged amino acid sequence of the Clostridium thermocellum wild-type licB amino acid sequence encoded by the nucleic acid sequence set forth in SEQ ID NO;
18, wherein the modified licB polypeptide comprises original N-terminal and C-terminal regions that correspond in sequence to N- and C-terminal polypeptides obtained by splitting the amino acid sequence encoded by SEQ ID NO;
18 within the loop region defined by amino acid residues 82-94 of the amino acid sequence encoded by SEQ ID NO;
18, wherein the modified licB polypeptide comprises the original C-terminal region fused upstream of the original N-terminal region, and wherein the recombinant carrier molecule optionally has a heterologous polypeptide comprising a therapeutic polypeptide or a disease-associated epitope fused thereto; and(b) growing said plant under conditions in which the nucleic acid is expressed and the carrier protein is produced. - View Dependent Claims (10, 11, 12, 13)
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Specification