PURIFICATION OF RNA-POLYMERASE ELONGATION COMPLEXES FOR CRYOELECTRON MICROSCOPY INVESTIGATION
Abstract
A procedure was developed for purification and preparation of a transcriptional stalled complex of Escherichia coli RNA polymerase for investigation by cryo-electron microscopy. The elongation complex has been concentrated on an affine monolayer formed by lipids bound to Ni ions. Additionally, the excess free RNA polymerase and unproductive complexes were removed on a heparin resin. The use of affinity grids with an immobilized monolayer of lipids helps to prevent aggregation of the particles on the grid surface. This technique can be used in the future to obtain a three-dimensional reconstruction of the EC+39 elongation complex.
About the Authors
O. V. Chertkov
Lomonosov Moscow State University
Russian Federation
Russian Federation
Department of Bioengineering, School of Biology,
Leninskiye gory 1–12, Moscow, 119234
M. G. Karlova
Lomonosov Moscow State University
Russian Federation
Russian Federation
Department of Bioengineering, School of Biology,
Leninskiye gory 1–12, Moscow, 119234
N. S. Gerasimova
Lomonosov Moscow State University
Russian Federation
Russian Federation
Department of Bioengineering, School of Biology,
Leninskiye gory 1–12, Moscow, 119234
O. S. Sokolova
Lomonosov Moscow State University
Russian Federation
Russian Federation
Department of Bioengineering, School of Biology,
Leninskiye gory 1–12, Moscow, 119234
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Review
For citations:
Chertkov O.V., Karlova M.G., Gerasimova N.S., Sokolova O.S. PURIFICATION OF RNA-POLYMERASE ELONGATION COMPLEXES FOR CRYOELECTRON MICROSCOPY INVESTIGATION. Vestnik Moskovskogo universiteta. Seriya 16. Biologiya. 2018;73(3):173-177. (In Russ.)
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