Reconstitution of Mcm4/6/7 using anion exchange chromatography. A) A portion of the indicated fractions from the Mono Q column were analyzed by SDS-PAGE (6%) and stained with Coomassie Brilliant Blue. The migration of size markers through the gel is indicated on the left. The migrations of Mcm4, Mcm6 and Mcm7 are indicated on the right. There is a minor contaminating band that migrates between Mcm6 and Mcm4 that is found in all of our preparations. B) ATP hydrolysis (black square) and DNA unwinding (white circle) by equal volumes of the indicated fractions were analyzed as described in the "Methods" section. C) A 50 μg portion of Mono Q fraction 67 was analyzed by gel filtration on a Superose 6 PC 3.2/30 column equilibrated in Buffer A with 100 mM NaCl. Shown here are Colloidal Coomassie Blue-stained SDS polyacrylamide gels (6%) of the indicated fractions. The vertical line marks the border between separate gels. The peak elutions of size standards are indicated at the bottom of the gel. The migration of size standards though the SDS gels is indicated on the left and of Mcm4, Mcm6 and Mcm7 on the right.
Ma et al. BMC Biochemistry 2010 11:37 doi:10.1186/1471-2091-11-37