Exploring the effects of active site constraints on hiv-1 reverse transcriptase dna polymerase fidelity*

HIGHLIGHTS

• who: Janina Cramer from the Proteins-Recombinant heterodimeric wild-type and M V mutant , RTs were expressed in Ecoli and purified as described before (36, ). Enzyme concentrations were routinely determined using an extinction coefficient at , nm of, M⫺, cm⫺1. Buffers-All experiments were carried out at , °C in a buffer containing , mM Tris-HCl, pH, .0, mM MgCl2, and , mM KCl. Annealing buffer consisted of , mM Tris-HCl, pH, .5, and , mM NaCl. Modified Thymidine, ⬘-Triphosphates-4⬘-Modified thymidine, ⬘triphosphates (TRTP) were synthesized as described previously (26). Oligonucleotides-Oligodeoxynucleotides were purchased from a commercial supplier and . . .
  

   

  

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