Modular construction of plasmids through ligation-free assembly of vector components with oligonucleotide linkers.
We have developed a modular method of plasmid construction that can join multiple DNA components in a single reaction. A nicking enzyme is used to create 5' and 3' overhangs on PCR-generated DNA components. Without the use of ligase or restriction enzymes, components are joined using oligonucleotide linkers that recognize the overhangs. By specifying the sequences of the linkers, desired components can be assembled in any combination and order to generate different plasmid vectors.
Vroom
Jonathan A
JA
Department of Chemical Engineering, Stanford University, Stanford, CA 94305, USA.
Wang
Clifford L
CL
D001483:Base Sequence; D003001:Cloning, Molecular; D008969:Molecular Sequence Data; D009841:Oligonucleotides / Q000737:chemistry / Q000235:genetics*; D010957:Plasmids
D009841:Oligonucleotides