Bacteriophage T7 DNA ligase. Overexpression, purification, crystallization, and characterization.

@article{Doherty1996BacteriophageTD,
  title={Bacteriophage T7 DNA ligase. Overexpression, purification, crystallization, and characterization.},
  author={Aidan J. Doherty and Stephen Ashford and Hosahalli S. Subramanya and Dale B. Wigley},
  journal={The Journal of biological chemistry},
  year={1996},
  volume={271 19},
  pages={11083-9}
}
The bacteriophage T7 DNA ligase gene was amplified using polymerase chain reaction-based methods and cloned into a T7 promoter-based expression vector. The protein was overexpressed to greater than 15% of total soluble protein and purified to homogeneity, yielding 60-70 mg of protein per liter of bacterial culture. An initial physical and biochemical characterization of the enzyme reveals that it exists as a monomer and can ligate nicked, cohesive, and blunt-ended DNA fragments. Inhibition of… CONTINUE READING

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