Replication of nucleic acids using single-strand DNA binding proteins

摘要

Single-strand DNA binding proteins have been found to increase the efficiency of strand-displacement replication of long targets, and when added to amplification reactions such as SDA, they significantly improve the amplification efficiency of targets which are otherwise too long to be efficiently amplified. The gp32 protein of T4 has been shown in SDA to produce about 10⁶ to 10⁸-fold amplification in 15-20 min. for targets about 800-1,000 bp in length. In the absence of gp32 there is essentially no detectable amplification for targets of this length. Targets about 400-500 bp in length are amplifiable about a billion-fold in the presence of gp32, representing an amplification factor approximately equivalent to that observed for targets less than 100 bp in length in conventional SDA reactions in the absence of SSBs. Other single-strand DNA binding proteins have also been shown to improve replication and amplification efficiency for long targets.