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. 2001 Oct 15;15(20):2730–2740. doi: 10.1101/gad.932201

Figure 1.

Figure 1

Expression and purification of the Mms4–Mus81 complex. (A) MMS4 and MUS81 were subcloned downstream of the T7 promoters of pET28a and pET11a, respectively, and further combined to yield a single plasmid, pNJ6220, carrying the indicated expression cassettes. (B) An extract from induced E. coli cells carrying plasmid pNJ6220 was applied to a phosphocellulose column, washed, and eluted with the indicated salt gradient. Column fractions were then resolved by SDS-PAGE, Western blotted, and probed with antiserum against Mms4 or Mus81 as indicated at right. (L) Load; (FT) flow through. (C) Fractions containing immunoreactive material in B were pooled, applied to a Ni-agarose column, washed, and eluted with buffer containing the indicated concentrations of imidazole. Fractions were Western blotted and probed with the indicated anti-serum. (D) Fractions in C were resolved by SDS-PAGE and stained with silver. (M) Molecular weight markers.