Method for genome mining for secreted protein genes
AssigneeNovozymes A/S
InventorsPreben Nielsen, Birthe Ravn Jørgensen
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Abstract
The present invention relates to a method for screening for compounds secreted by an organism, comprising (a) raising antibodies against secreted products of a donor organism, (b) providing a gene library from the donor organism, (c) cloning the gene library into a suitable host organism, (d) expressing the cloned genes in the host organism, and (e) detecting positive clones, which upon expression of the cloned genes secretes a compound, using the antibodies of (a) to identify such positive clones. The invention also relates to compounds, nucleotide sequences and microorganisms identifiable by said method.
Claims12
A method for screening for compounds secreted by a microorganism, comprising:
- (a) cultivating the microorganism to produce a supernatant or a supernatant including surface bound compounds of the microorganism which comprises at least 2 secreted products,
- (b) preparing a mixture of antibodies by immunizing an animal with the supernatant comprising at least 2 secreted products and isolating the mixture of antibodies formed in the animal,
- (c) providing a gene library from the microorganism,
- (d) cloning the gene library into a suitable host organism,
- (e) screening clones of the host organism using isolated antibody mixture to identify positive clones expressing a cloned gene encoding a secreted compound, and
- (f) screening positive clones for peptides or proteins having a desired function or subjecting positive clones to nucleotide sequencing in order to identify genes encoding compounds of interest.
A method for screening for compounds secreted by a microorganism, comprising:
- (a) cultivating the microorganism to produce a supernatant or a supernatant including surface bound compounds of the organism which comprises at least 2 secreted products,
- (b) raising antibodies against all compounds in the supernatant or the supernatant including surface bound compounds,
- (c) providing a gene library from the organism,
- (d) cloning the gene library into a suitable host organism,
- (e) screening clones of the host organism using isolated antibody mixture to identify positive clones expressing a cloned gene encoding a secreted compound, and
- (f) screening positive clones for peptides or proteins having a desired function or subjecting positive clones to nucleotide sequencing in order to identify genes encoding compounds of interest.
Dependencies are inferred from the claim text.