A New EBS2b-IBS2b Base Paring (A −8 /T −8 ) Improved the Gene-Targeting Efficiency of Thermotargetron in Escherichia coli

The randomized base pairing in the interval of IBS2 and IBS1 of Tel3c/4c intron (−8 and −7 sites) in Thermotargetron (TMT) results in a low success rate and gene-targeting efficiency in bacteria. In the present work, we constructed a randomized gene-targeting plasmids pool (RGPP) to study whether there is a base preference in target sequences.