Title:

Joining Forces: Engineering a High-Efficiency DNA Ligase for Tunable Biomaterials

DNA ligases are essential enzymes in biotechnology and materials science, but commercial sources can be expensive and may not meet specific research needs. In this study, we successfully engineered a high-efficiency DNA ligase using a cost-effective and scalable expression and purification protocol. The ligase was expressed in BL21 E. coli cells using a selective growth medium, and purification was performed using Immobilized Metal-Chelate Affinity Chromatography (IMAC) with a 6-His tag. Our results show that the engineered ligase has a higher ligation efficiency and speed compared to commercial T4 ligase when ligating linear DNA strands. Moreover, our expression and purification protocol yielded a significant amount of highly pure protein in a single-step purification. This work demonstrates the feasibility of producing a customizable and cost-effective DNA ligase for use in biopolymer materials research.

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