Cloning Strategies Using a Third Irrelevant Enzyme Site (TIES) to Overcome Certain Cloning Problems

Abstract

We describe a cloning strategy, third irrelevant enzyme site (TIES), based on threecomponent assembly ligation reactions to overcome seven types of problematical cloning reactions. Implementation of the TIES strategy has made it possible to obtain the desired recombinant where the corresponding approach using conventional recombinant DNA methodologies and twocomponent ligations failed to give the desire product.