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Figure 1 Design of the bidirectional ligase maxizyme. (a) The DNA ligase was designed to be a binary (two black strands) enzyme with two catalytic domains fused by a common stem structure. When the binary strands associate to form the correct structure, the catalytic domains are formed and are capable of ligating two DNA substrates (green and purple). (b) An effector DNA (red) can specifically base pair with the binary enzyme, stimulating the correct folded structure and catalyzing the ligation of two substrates on the opposite end. In this way the effector oligonucleotide is recoded into a new oligonucleotide ligation product. For convention, we refer to ligation of substrates on the ‘bottom’ and on the ‘top’ of the enzyme.
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