Efficient DNA ligation in DNA-RNA hybrid helices by Chlorella virus DNA ligase

@inproceedings{Lohman2014EfficientDL,
  title={Efficient DNA ligation in DNA-RNA hybrid helices by Chlorella virus DNA ligase},
  author={Gregory J S Lohman and Yi Zheng Zhang and Alexander M. Zhelkovsky and Eric J Cantor and Thomas C Evans},
  booktitle={Nucleic acids research},
  year={2014}
}
Single-stranded DNA molecules (ssDNA) annealed to an RNA splint are notoriously poor substrates for DNA ligases. Herein we report the unexpectedly efficient ligation of RNA-splinted DNA by Chlorella virus DNA ligase (PBCV-1 DNA ligase). PBCV-1 DNA ligase ligated ssDNA splinted by RNA with kcat ≈ 8 x 10(-3) s(-1) and K(M) < 1 nM at 25 °C under conditions where T4 DNA ligase produced only 5'-adenylylated DNA with a 20-fold lower kcat and a K(M) ≈ 300 nM. The rate of ligation increased with… CONTINUE READING
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