title: Single restriction enzyme-assisted megaprimer polymerase chain reaction to fuse two DNA sequences on separate cloning vectors.
abstract: We describe a simple and versatile method to fuse two DNA sequences on separate cloning vectors in a single polymerase chain reaction (PCR). The method, termed restriction enzyme-assisted megaprimer PCR (REM-PCR), requires that the two cloning vectors share a common sequence and that the DNA sequences to be fused are cloned in the same orientation with respect to the common sequence. Fusion of the two sequences is achieved by mutual priming at the common sequence between two DNA fragments that were generated by restriction enzyme and linearly amplified by repetitive priming in the PCR reaction mixture.
authors: Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology, Higashi, Tsukuba, Ibaraki, Japan.
authors: Nobuhisa
authors: N
authors: Umeki
authors: Taro Q P
authors: TQ
authors: Uyeda
mesh_terms: D001483:Base Sequence; D003001:Cloning, Molecular; D004247:DNA / Q000737:chemistry; D004262:DNA Restriction Enzymes; D005822:Genetic Vectors / Q000737:chemistry*; D016133:Polymerase Chain Reaction / Q000379:methods*
chemical_list: D004247:DNA; D004262:DNA Restriction Enzymes