Novel Approach to the Ligation of Single-Stranded DNA Fragments by T4 DNA Ligase—DNA Mobile Multiple-Restriction Fragments: “UNI-LINKERS” for Cloning of Genes

Abstract A group of uniquely designed single-stranded oligodeoxyribo-nucleotides that form hairpin loops were synthesized. These oligonucleotides can be ligated to other synthetic single-stranded fragents differing in length and design without the need for external annealing templates. A novel approach to building a limitless variety of mobile multiple-restriction DNA fragments termed “uni-linkers” and which are open only at one end, is described. The latter were used as “multi-stage” linkers in cloning experiments. The cloning of the cDNA for a carrot proline-rich protein, DC-5, into a plant vector is presented as an example. Other possible applications are also discussed.