Recombinant DNA is a DNA molecule formed by joining sequence segments into a new genetic arrangement. Complementary ends can align the constituent fragments, and polymerase and ligase activities complete a continuous DNA backbone [1]. Constructs can combine promoters, coding sequences, and termination elements into single or multiple expression units, with the orientation and boundaries of each segment specified during assembly [2]. Restriction-based joining uses recognition and cleavage properties that generate compatible fragment ends; controlled methylation can provide additional specificity for cleavage and ligation [3]. Hierarchical assembly connects intermediate constructs into larger molecules while preserving the intended sequence organization [4]. The resulting molecule carries the combined nucleotide information, including the designed junctions and any replication or selection elements included in the construct [5].
- DNA assembly
- plasmid constructs
- sequence junctions
- expression constructs
References
- Daniel G Gibson; Lei Young; Ray-Yuan Chuang; J Craig Venter; Clyde A Hutchison; Hamilton O Smith; Enzymatic Assembly of DNA Molecules up to Several Hundred Kilobases. Nat. Methods 2009, 6, 343-345, 10.1038/nmeth.1318.
- George M Taylor; Paweł M Mordaka; John T Heap; Start-Stop Assembly: a Functionally Scarless DNA Assembly System Optimized for Metabolic Engineering. Nucleic Acids Res. 2018, 47, e17-e17, 10.1093/nar/gky1182.
- Wei-Hua Chen; Zhong-Jun Qin; Jin Wang; Guo-Ping Zhao; The MASTER (methylation-assisted tailorable ends rational) Ligation Method for Seamless DNA Assembly. Nucleic Acids Res. 2013, 41, e93-e93, 10.1093/nar/gkt122.
- Da Lin; Christopher A O’cAllaghan; MetClo: Methylase-assisted Hierarchical DNA Assembly Using a Single Type IIS Restriction Enzyme. Nucleic Acids Res. 2018, 46, e113-e113, 10.1093/nar/gky596.
- Fang Ba; Qing Sun; Site-specificRecombinases and Their Essential Role in Synthetic Biology. Biotechnol. Adv. 2026, 92, 109005, 10.1016/j.biotechadv.2026.109005.