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. 2021 Jan 14;8:553474. doi: 10.3389/fbioe.2020.553474

FIGURE 2.

FIGURE 2

Strand-displacement by mutant enzymes with D732N. (A) Half-dumbbell test. Catalyzed DNA synthesis begins at the primer (arrow) and proceeds to and through the loop and back again, only if the polymerase is capable of strand displacement, to make a product 2.4 kb in size. The two D732N mutant versions of Taq (full-length A111 and Klentaq1) are marked with asterisk (*). Bst pol was expected to be able to strand-displace, and was included as a positive control. We explain unextended 1.2 kb template as due to inefficiency of the half-dumbbell construction at unknown point(s) during its preparation. (B) Discovery of LAMP ability. A multiply mutant form of Taq pol carrying D732N, named KTflnC4RR (lanes KT*, see text) was tested compared Manta (lanes M), a brand of the large fragment of Bst DNA polymerase (Enzymatics). The banded amplification product which is typical for LAMP appeared at 3 h incubation at 60°C for both enzymes but only for KT* enzyme at 70°C. The middle lane size standards were 1, 2, 4 kb.