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. Author manuscript; available in PMC: 2015 Aug 24.
Published in final edited form as: J Biol Chem. 2001 Mar 15;276(22):19141–19149. doi: 10.1074/jbc.M100329200

Fig. 2. Experimental strategy for construction of circularly permuted ALAS variants-containing library.

Fig. 2

A murine erythroid ALAS cDNA obtained by digestion of pAC10 vector with SalI (see “Experimental Procedures”) was circularized, and the circularized DNA was subsequently treated with RQ1 DNase. The randomly linearized ALAS-encoding fragments were repaired and blunt-ended with T4 DNA polymerase Klenow fragment and subcloned into the pAC9 expression vector to yield a library of circularly permuted ALAS variants (see “Experimental Procedures” for details).