Fig. 3. The efficiency and accuracy of pTk-RadA splicing is radically improved at high temperature or through addition of ssDNA in vitro.
(A and B) Native PAGE and quantification of fluorescent signals reveal the impact of temperature on the efficiency of intein excision of Tk-RadAΔ286–858. (C) SDS-PAGE of purified pTk-RadAΔ286–585 before incubation (T0) without (−) and with (+) an ssDNA cofactor that markedly increases the efficiency and accuracy of intein splicing and extein ligation. The lane labeled M contains proteins of known molecular weights measured in kilodaltons.