To ascertain the hydrolytic nature of the DNA cleavage reaction
mediated by complex 3 DNA religation experiment was performed
in which supercoiled pBR322 DNA was treated with T4 ligase
enzyme and subjected to gel electrophoresis [35]. The complex 3
yielded nicked DNA which was religated by using T4 DNA ligase
enzyme; nicked form (FormII) was religated to a large extent in the presence of T4 ligase enzyme in comparison to control DNA alone in
supercoiled form