High throughput screening method for identifying molecules having biocidal function
OwnerXiongying Cheng, Illimar Altosaarinventors
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Abstract
Novel methods for screening and cloning DNA sequences coding for biocidal molecules, and for identifying biocidal molecules involving use of viability staining assay are provided. The methods consist of construction of libraries of DNA from natural and synthetic sources, and expression of cloned DNA in surrogate hosts, identification of DNA clones biocidal to the host cells by viability assay and isolation of the DNA sequences coding for biocidal molecules. Further provided are methods of identifying the biocidal molecules and their use.
Claims15
A method of identifying nucleic acid sequence encoding molecule having biocidal function comprising the steps of:
- a. constructing a library of nucleic acid molecules, isolated from an organism or part thereof, or synthesized chemically, each of the nucleic acid molecules operably linked to an inducible promoter sequence in a vector,
- b. inducing the library into host cells to produce transformed cells;
- c. growing the transformed cells in the absence of an inducer,
- d. adding the inducer and growing the transformed cells to express each of the nucleic acid molecules in the library to produce a library of induced colonies or cells;
- e. staining the induced colonies or cells with one or more than one dye;
- f. determining the viability of the induced colonies or cells, and identifying colonies with reduced or lost cell viability;
- and
- g. isolating the nucleic acid sequence from the colonies or cells with reduced or lost cell viability.
Dependencies are inferred from the claim text.