Patent ID: 6913886

Claim:
A method for selecting a polynucleotide encoding a prey polypeptide, said prey polypeptide being able to interact with a bait polypeptide, said method comprising: (A) providing a yeast cell mixture comprising a first population of yeast cells, which are not in a colony, containing at least one first haploid recombinant yeast cell containing a polynucleotide encoding said bait polypeptide (bait polynucleotide) and a second population of yeast cells, which are not in a colony, containing at least one second haploid recombinant yeast cell containing a polynucleotide encoding said prey polypeptide (prey polynucleotide); (B) subjecting said polynucleotide encoding said bait polypeptide to a two-hybrid screening method, wherein said two-hybrid screening method comprises a step of mating for a maximum of 5 hours, without replica plating, said at least one first haploid recombinant yeast cell containing said bait polynucleotide with said second haploid recombinant yeast cell containing said prey polynucleotide, provided that one haploid yeast cell among said first haploid recombinant yeast cell or said second haploid recombinant yeast cell also contains at least one detectable gene that is activated by a polypeptide including a transcriptional activation domain; and (C) selecting the recombinant diploid yeast cell obtained at step (B) for which said detectable gene has been expressed to a degree greater than expression in the absence of interaction between said bait polypeptide and said prey polypeptide.