The Application Of Type Ⅱ Restriction Endonuclease PstI To Synthetic Biology

Here we have a research on the influence of PstI restriction endonuclease on the host genomes. The PstI protein ,which had been expressed by RosettaTM transfromed plasmid with pstI gene under the induction of IPTG in the final concentration 1 mM in 30 oC, was proved to have the same function as normal type II restriction endonuclease by enzyme digestion experiment in vitro. This study had found that PstI restriction endonuclease could digest the host genomes which would be degraded by DNase latter, and attained bacteria shell without genomes finally. Our results revealed RosettaTM genomes were totally degraded after inducing three hours using DAPI dyeing method with laser scanning confocal microscope. Accordingly, by this means we can acquire a protein synthetic system without genomes and an intact bacteria shell.