摘要
In this note, we report a novel and efficient three primers PCR (TP-PCR) method to rapidly generate recombinant DNA molecule at precise junction between two arbitrary DNA fragments. TP-PCR method is characterized by its reaction system with two templates and three primers, which can produce a recombinant DNA molecule in one PCR reaction. The main advantages of this method are the independence of sequences at the recombination site, the rapid-ness, and the easy establishment of adequate conditions. This method has been successfully applied to constructing a fusion protein gene, sck gene.
In this note, we report a novel and efficient three primers PCR (TP-PCR) method to rapidly generate recombinant DNA molecule at precise junction between two arbitrary DNA fragments. TP-PCR method is characterized by its reaction system with two templates and three primers, which can produce a recombinant DNA molecule in one PCR reaction. The main advantages of this method are the independence of sequences at the recombination site, the rapidness, and the easy establishment of adequate conditions. This method has been successfully applied to constructing a fusion protein gene, sck gene.
基金
This work was supported by the National High Science and Technology Program "863" (Grant Nos. 2001AA212041 and 2001AA222251)
the National Natural Science Foundation of China (Grant No. 39989001)
the National Special Program for Research and Industr
