Gene cluster screening of clones having DNA from mixed populations of organisms
View Patent ↗Disclosed is a process of screening clones having DNA from an uncultivated microorganism for a specified protein, e.g. enzyme, activity by screening for a specified protein, e.g. enzyme, activity in a library of clones prepared by (i) recovering DNA from a DNA population derived from at least one uncultivated microorganism; and (ii) transforming a host with recovered DNA to produce a library of clones which is screened for the specified protein, e.g. enzyme, activity.
1. A method for identifying a polyketide synthase gene cluster of interest comprising:
culturing a gene expression library comprising a pool of expression constructs, each expression construct comprising an f-factor based vector containing one or more suitably-sized naturally-occurring genomic DNA fragments, wherein the genomic DNA fragments in the pool of expression constructs are directly obtained from a plurality of species of uncultivated donor microorganisms and wherein the genomic DNA fragments are operably-associated with one or more regulatory regions that drives expression of genes encoded by the genomic DNA fragments in an appropriate host organism; and
detecting a naturally-occurring polyketide synthase gene cluster contained in one or more of the naturally-occurring genomic DNA fragments.
2. The method of claim 1 , wherein the host organism is a prokaryotic cell.
3. The method of claim 1 , wherein the host organism is a eukaryotic cell.
4. The method of claim 1 , wherein the donor organism is a fungal cell.
5. The method of claim 1 , wherein the donor organism are prokaryotic cells.
6. The method of claim 1 , wherein the donor organism are eukaryotic cells.
7. The method of claim 1 , wherein the donor organism are fungal cells.
8. The method of claim 1 , wherein the genomic DNA fragments are operably associated with their native regulatory region(s).