Gene cluster for ramoplanin biosynthesis
AssigneeEcopia BioSciences, Inc.
InventorsChris M. Farnet, Emmanuel Zazopoulos, Alfredo Staffa
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Abstract
The present invention relates to isolated genetic sequences encoding proteins which direct the biosynthesis of the antibiotic ramoplanin in microorganisms. The isolated biosynthetic gene cluster serves as a substrate for bioengineering of antibiotic structures.
Claims24
An isolated nucleic acid comprising a nucleic acid sequence selected from the group consisting of:
- (a) a nucleic acid encoding any of ramoplanin ORFs 1 to 33 (SEQ ID NOS:
- 2 to 34);
- (b) a nucleic acid encoding a polypeptide encoded by any of ramoplanin ORFs 1 to 33 (SEQ ID NOS:
- 2 to 34);
- (c) a nucleic acid amplified by polymerase chain reaction (PCR) using primer pairs that amplify any of ramoplanin ORFs 1 to 33 (SEQ ID NOS:
- 2 to 34).
An isolated nucleic acid comprising a nucleic acid sequence selected from the group consisting of:
- (a) a nucleic acid encoding any of ramoplanin ORFs 4, 5, 9 to 19, 22 to 26, 29, 30 and 31 (SEQ ID NOS:
- 5, 6, 10 to 20, 23 to 27, 30, 31 and 32);
- (b) a nucleic acid encoding a polypeptide encoded by any of ramoplanin ORFs 4, 5, 9 to 19, 22 to 26, 29, 30 and 31 (SEQ ID NOS:
- 5, 6, 10 to 20, 23 to 27, 30, 31 and 32);
- (c) a nucleic acid encoding a polypeptide that is at least 75% identical in amino acid sequence to a polypeptide of ramoplanin ORFs 4, 5, 9 to 19, 22 to 26, 29, 30 and 31 (SEQ ID NOS:
- 5, 6, 10 to 20, 23 to 27, 30, 31 and 32).
An isolated nucleic acid comprising a nucleic acid that hybridizes under stringent conditions to any one of ORFs 1 to 32 of the ramoplanin biosynthesis gene cluster (SEQ ID NOS:
- 2 to 33) and can substitute for the ORF to which it specifically hybridizes to direct the synthesis of a ramoplanin.
An isolated nucleic acid comprising a nucleic acid that hybridizes under stringent conditions to any one of ORFs 4, 5, 9 to 19, 22 to 26, 29, 30 and 31 (SEQ ID NOS:
- 5, 6, 10 to 20, 23 to 27, 30, 31 and 32) of the ramoplanin biosynthesis gene cluster and can substitute for the ORF to which it specifically hybridizes to direct the synthesis of a ramoplanin.
An isolated gene cluster comprising ORFs encoding polypeptides sufficient to direct the synthesis of a ramoplanin or a ramoplanin analog.
The isolated gene cluster of claim 12 wherein the gene cluster is present in a bacterium.
An isolated polypeptide comprising a polypeptide sequence selected from any one of:
- (a) a polypeptide of any one of ORFs 1 to 32 (SEQ ID NOS:
- 2 to 33);
- and
- (b) a polypeptide which is at least 75% identical in amino acid sequence to a polypeptide of any one of ORFs 1 to 32 (SEQ ID NOS:
- 2 to 33).
Dependencies are inferred from the claim text.