Methods and compositions for directed gene assembly

Abstract

The present invention is directed to methods and compositions for use of homologous recombination for directed evolution, gene reassembly, and directed mutagenesis. One aspect of the present invention relates to methods for use of bacterial conjugative transfer and homologous recombination for directed evolution, gene reassembly, and directed mutagenesis. Another aspect of the present invention relates to compositions for use in or produced by the methods of the present invention, including libraries, archives and databases.

1 .- 39 . (canceled)  
     
     
         40 . A donor library comprising a plurality of donor vectors, said donor vectors comprising in the following order from 5′ to 3′: a first donor DNA sequence and a second donor DNA sequence, such that said first donor DNA sequence is homologous to a first target DNA sequence of a target gene of interest, and a second donor DNA sequence is homologous to a second target DNA sequence of a target gene of interest, wherein said donor vectors are suicide vectors.  
     
     
         41 . The donor library of  claim 40  wherein the members are arrayed.  
     
     
         42 . The donor library of  claim 41  wherein the members are arrayed in an 8×12, 16×24, or a 32×48 matrix.  
     
     
         43 . (canceled)  
     
     
         44 . (canceled)