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Updated: Sep 29, 2026

Chemical Triphosphorylation of Oligonucleotides
Published on: June 2, 2022
Purification, crystallization and initial crystallographic analysis of RNA polymerase holoenzyme from Thermus
Marina N Vassylyeva1, Jookyung Lee, Shun Ichi Sekine
1Genomic Sciences Center (RIKEN), 2-1 Hirosawa, Wako, Saitama 351-0198, Japan.
Abstract:
RNA polymerase holoenzyme from Thermus thermophilus, consisting of six protein subunits (alpha(2), beta, beta', omega and sigma(70)) and having a total molecular mass of about 450 kDa, was purified and crystallized by the hanging-drop vapour-diffusion technique under mild near-physiological conditions. The crystals diffract beyond 3 A resolution. Careful analysis of diffraction data revealed that the crystals belong to space group P3(2), with unit-cell parameters a = b = 236.35, c = 249.04 A, and have perfect twinning along the threefold axis. A complete data set at 3 A resolution was collected and an unambiguous molecular-replacement solution was found using the structure of T. aquaticus RNA polymerase core enzyme as a search model. The refinement of structure and model building of the sigma(70) subunit is now in progress.
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