Methods and nucleic acid sequences for the expression of the cellulose synthase operon
AssigneeCetus Corporation
InventorsArie Ben-Bassat, Roger D. Calhoon, Anna L. Fear, David H. Gelfand, James H. Meade, Rony Tal, Hing Wong, Moshe Benziman
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Abstract
Nucleic acid sequences encoding the bacterial cellulose synthase operon derived from Acetobacter are disclosed. Methods for isolating the genes, vectors containing the genes, and transformed hosts useful for the expression of recombinant bacterial cellulose synthase or production of cellulose are also described.
Claims53
An isolated polynucleotide acid segment consisting essentially of contiguous nucleic acid sequences encoding a bacterial cellulose synthase operon (SEQ ID No:
- 1), wherein said polynucleotide acid segment is capable of hybridizing to one or more of the four genes of Sequence ID No:
- 1 under stringent conditions.
An isolated polynucleotide substantially corresponding to the nucleotide segment consisting essentially of the nucleic acid sequence encoding cellulose synthase A (SEQ ID No:
- 3) wherein said polynucleotide acid segment is capable of hybridizing to the polynucleotide of Sequence ID No:
- 3 under stringent conditions.
An isolated polynucleotide substantially corresponding to the nucleotide segment consisting essentially of the nucleic acid sequence encoding cellulose synthase B (SEQ ID No:
- 4) wherein said polynucleotide acid segment is capable of hybridizing to the polynucleotide of Sequence ID No:
- 4 under stringent conditions.
An isolated polynucleotide substantially corresponding to the nucleotide segment consisting essentially of the nucleic acid sequence encoding cellulose synthase C (SEQ ID No:
- 5) wherein said polynucleotide acid segment is capable of hybridizing to the polynucleotide of Sequence ID No:
- 5 under stringent conditions.
An isolated, polynucleotide substantially corresponding to the nucleotide segment consisting essentially of the nucleic acid sequence encoding cellulose synthase D (SEQ ID No:
- 6) wherein said polynucleotide of Sequence ID No:
- 6 under stringent conditions.
A method for increasing cellulose production in a recombinant microorganism, which method comprises:
- a) transforming a strain of a cellulose producing species of bacteria with a vector comprising at least one gene derived from the cellulose synthase operon, selected from the group consisting of cellulose synthase genes B, C, D, taken singly, and mixtures of genes A, B, C, and D;
- and
- (b) culturing said transformed microorganism under conditions suitable for the production of cellulose;
- and
- (c) recovering the cellulose.
A recombinant DNA vector comprising:
- a) a functional Acetobacter origin of replication-containing fragment of plasmid p824, and
- b) one or more DNA segments that convey resistance to at least one antibiotic when transformed into a sensitive host cell that is susceptible to transformation, cell division and culture.
An isolated protein having the amino acid sequence of an Acetobacter cellulose synthase B (SEQ ID No:2 residues 755-1556) and allelic variants thereof.
An isolated protein having the amino acid sequence of an Acetobacter cellulose synthase A (SEQ ID No:2 residue 1-754) and allelic variants thereof.
An isolated protein having the amino acid sequence of an Acetobacter cellulose synthase C (SEQ ID No:2 residues 1557-2875) and allelic variants thereof.
An isolated protein having the amino acid sequence of an Acetobacter cellulose synthase D (SEQ ID No:2 residues 2876-3031) and allelic variants thereof.
Dependencies are inferred from the claim text.