Figure 2

DNA ligation and adenylyltransferase activities of the putative recombinant DNA ligases. A. LigA wt and LigA K128A were incubated with [³²P]-NAD⁺ and adenylyltransferase activity was detected by SDS-PAGE. Protein bands were visualized with Coomassie Blue R250 (left) and by autoradiography (right). B. DRB0100 wt and K40A were incubated with α-[³²P]-ATP. Protein-AMP complexes and free α-[³²P]-ATP were separated by SDS-PAGE. Proteins were stained with Coomassie Blue R250 (left) and detected by autoradiography (right). C. Titration of LigA wt and LigA K128A on a nicked DNA substrate. Indicated amounts of LigA wt and LigA K128A were incubated with the DNA substrate as described in the Methods section. [³²P]-labelled DNA oligonucleotides were visualized by autoradiography. D. Quantification of three independent experiments as shown in C. Error bars are given as the standard error of the mean.