Overview of StrataClone PCR Cloning Technology
This technology exploits the activities of topoisomerase I from Vaccinia virus and Cre recombinase from bacteriophage P1. In vivo, DNA topoisomerase I assists in DNA replication by relaxing and rejoining DNA strands. Topoisomerase I cleaves the phosphodiester backbone of DNA strands after the sequence 5´-CCCTT. it forms a covalent DNA–enzyme intermediate that conserves bond energy for religating the cleaved DNA back to the original strand. Once the covalent DNA–enzyme intermediate is formed, the religation reaction can also occur with a heterologous DNA acceptor.