Ribb
AssigneeSmithKline Beecham Corporation
InventorsMichael Terance Black, Lisa Kathleen Shilling, Robert King Stodola, Richard Lloyd Warren, Anna Lisa Kosmatka, Richard Oakley Nicholas, Leslie Marie Palmer, Michael Arthur Lonetto, Jason Craig Fedon, John Edward Hodgson, David Justin Charles Knowles
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Abstract
The invention provides ribB polypeptides and polynucleotides encoding ribB polypeptides and methods for producing such polypeptides by recombinant techniques. Also provided are methods for utilizing ribB polypeptides to screen for antibacterial compounds.
Claims16
An isolated polypeptide consisting of the amino acid sequence as set forth in SEQ ID NO:
- 2.
An isolated polypeptide encoded by an isolated first polynucleotide wherein the isolated first polynucleotide hybridizes under stringent conditions to a second polynucleotide which encodes the mature polypeptide of SEQ ID NO:
- 2;
- wherein stringent conditions comprise overnight incubation at 42° C. in a solution comprising:
- 50% formamide, 5×SSC (150 mM NaCl, 15 mM trisodium citrate), 50 mM sodium phosphate (pH7.6), 5×Denhardt's solution, 10% dextran sulfate, and 20 micrograms/ml denatured, sheared salmon sperm DNA, followed by washing the filters in 0.1×SSC at 65° C.;
- wherein the isolated polypeptide has riboflavin synthase enzymatic activity expressed by the DNA contained in NCIMB Deposit No. 40771.
An isolated polypeptide encoded by an isolated first polynucleotide wherein the isolated first polynucleotide hybridizes under stringent conditions to the polynucleotide sequence of SEQ ID NO:
- 1, wherein stringent conditions comprise overnight incubation at 42° C. in a solution comprising:
- 50% formamide, 5×SSC (150 mM NaCl, 15 mM trisodium citrate), 50 mM sodium phosphate (pH7.6), 5×Denhardt's solution, 10% dextran sulfate, and 20 micrograms/ml denatured, sheared salmon sperm DNA, followed by washing the filters in 0.1×SSC at 65° C.;
- wherein the isolated polypeptide comprises a sequence of at least 30 amino acids.
An isolated polypeptide comprising a polypeptide sequence selected from the group consisting of:
- (a) a first sequence which is SEQ ID NO:
- 2, and
- (b) a second sequence comprising at least 50 consecutive amino acids of SEQ ID NO:
- 2.
- wherein the isolated polypeptide is effective to induce antibodies to a polypeptide having the sequence of SEQ ID NO:2.
An isolated polypeptide comprising a polypeptide consisting of SEQ ID NO:
- 2 and a heterologous amino acid sequence fused to the polypeptide.
An isolated polypeptide comprising a polypeptide, wherein the polypeptide consists of an amino acid sequence identical to SEQ ID NO:
- 2 except that, over the entire length corresponding to SEQ ID NO:
- 2, the amino acid sequence has a substitution, deletion or insertion of one amino acid, wherein the isolated polypeptide has riboflavin synthase enzymatic activity.
An isolated polypeptide encoded by an isolated first polynucleotide wherein the isolated first polynucleotide hybridizes under stringent conditions to the polynucleotide sequence of SEQ ID NO:
- 3, wherein stringent conditions comprise overnight incubation at 42° C. in a solution comprising:
- 50% formamide, 5×SSC (150 mM NaCl, 15 mM trisodium citrate), 50 mM sodium phosphate (pH7.6), 5×Denhardt's solution, 10% dextran sulfate, and 20 micrograms/ml denatured, sheared salmon sperm DNA, followed by washing the filters in 0.1×SSC at 65° C.;
- wherein the isolated polypeptide comprises a polypeptide sequence of SEQ ID NO:4.
Dependencies are inferred from the claim text.