A simple and highly efficient cloning method that employs PCR to directly create a fusion between insert and vector.
We have developed a very simple procedure for gene cloning using compound primers. This approach permits the insertion of a DNA fragment in a single step using a PCR-based cloning protocol, which employs annealing rather than ligation to create the recombinant in the plasmid. This method has been tested successfully to clone a Phalaenopsis gene coding for P450. A potential application of this protocol for constructing a cDNA library is also proposed.
Su
Vincent
V
Department of Life Science, National Tsing Hua University, Hsin-Chu 30055, Taiwan.
Hsu
Ban-Dar
BD
D003001:Cloning, Molecular / Q000379:methods*; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics*; D018744:DNA, Plant / Q000235:genetics*; D015723:Gene Library; D029595:Orchidaceae / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*
D018744:DNA, Plant; D003577:Cytochrome P-450 Enzyme System