Method for preselecting recombinant clones containing a specific nucleic acid sequence and subsequent transformation with preselected clones

摘要

A method for preselecting a nucleic acid sequence of interest is provided. The method comprises immobilizing an affinity molecule to a solid support matrix, wherein the affinity molecule is specific for a hapten or target molecule; hybridization of a "hook" comprised of either an oligonucleotide (complementary to the sequence of interest) labeled with a hapten or target molecule to the target recombinant vector containing the nucleic acid sequence of interest; capturing of the vector- hook via the hapten or target molecule by the respective immobilized affinity molecule for which it has binding specificity; liberating the captured vector DNA by enzymatic digestion of either the hook, or of the immobilized affinity molecule; and adding and incubating competent host cells to promote the introduction of recombinant vector DNA into the competent host cells. Also disclosed are embodiments in which a) the hook is incorporated as part of the vector by hybridization to a denatured vector, followed by second strand synthesis to include the hook as part of the newly synthesized strand; and b) the hook is a protein that binds a genetic element within or adjacent to the nucleic acid sequence of interest, and the immobilized affinity molecule has binding specificity for the protein.