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[Music] the overall goal of this procedure is to prepare large oligomeric dna binding proteins such as mgm 101 using the maltose binding protein or mbp tagging strategy this is accomplished by first cloning the mgm 101 open reading frame downstream of mvp in the pmal c2e vector for expression in e coli the mbp mgm-101 fusion protein is then purified from e coli by amylose affinity chromatography mgm-101 is then released from mbp by proteolytic cleavage and separated from mbp by cation exchange chromatography finally the mgm-101 rings are purified to homogeneity by size exclusion chromatography ultimately approximately 0.8 milligrams of soluble mgm-101 from one liter of bacterial culture can be used for biochemical and structural characterization of this protein which is important for dna repair in mitochondria the main advantage of using a motors binding protein tagged version of mgm 101 is that this version is less toxic more soluble and more stable than the non-tagged version as a re
