HIGHLIGHTS
- who: Rajam S. Mani from the Bacterial PNKP expression and purification His-tagged human PNKP, PNKPWFX402, and the FHA domain of PNKP were expressed in Ecoli BL21(DE3)pLysS (Millipore, Etobicoke, Ontario, Canada), purified as described previously (10, , ), and kept in storage buffer (, mM TrisHCl, pH, .4, mM NaCl, mM MgCl2, and , mM dithiothreitol) at ⫺, °C. Bacterial , expression and purification The , cDNA encoding residues, (EB2) and, (BLB) incorporating a, terminal hexahistidine tag were cloned into the pET28a vector (Millipore) using a restriction-free cloning procedure (46). Briefly, in the first step a regular PCR amplification was performed . . .
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