Temiakov Dmitry, Tahirov Tahir H, Anikin Michael, McAllister William T, Vassylyev Dmitry G, Yokoyama Shigeyuki
Morse Institute of Molecular Genetics, Department of Microbiology and Immunology, SUNY Health Science Center at Brooklyn, 450 Clarkson Avenue, Brooklyn, New York 11203-2098, USA.
Acta Crystallogr D Biol Crystallogr. 2003 Jan;59(Pt 1):185-7. doi: 10.1107/s0907444902019777. Epub 2002 Dec 19.
Stable transcription-elongation complexes consisting of T7 RNA polymerase (molecular mass 99 kDa) in association with a nucleic acid scaffold consisting of an 8 bp RNA-DNA hybrid and 10 bp of downstream DNA were assembled and crystallized by the sitting-drop vapour-diffusion technique under near-physiological conditions. The crystals diffract beyond 2.6 A resolution and belong to space group P1, with unit-cell parameters a = 79.91, b = 84.97, c = 202 A, alpha = 90.36, beta = 92.97, gamma = 109.94 degrees. An unambiguous molecular-replacement solution was found using the C-terminal portion of the T7 RNA polymerase structure from the early initiation complex as a search model. Model building and structure refinement are now in progress.
在接近生理条件下，通过坐滴气相扩散技术组装并结晶了由T7 RNA聚合酶（分子量99 kDa）与由8 bp RNA-DNA杂交体和10 bp下游DNA组成的核酸支架形成的稳定转录延伸复合物。这些晶体的衍射分辨率超过2.6 Å，属于空间群P1，晶胞参数为a = 79.91，b = 84.97，c = 202 Å，α = 90.36，β = 92.97，γ = 109.94°。使用来自早期起始复合物的T7 RNA聚合酶结构的C末端部分作为搜索模型，找到了明确的分子置换解决方案。目前正在进行模型构建和结构优化。
