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. 2014 Jul;20(7):1023–1034. doi: 10.1261/rna.043877.113

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© 2014 Otero et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society

This article is distributed exclusively by the RNA Society for the first 12 months after the full-issue publication date (see http://rnajournal.cshlp.org/site/misc/terms.xhtml). After 12 months, it is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/.

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FIGURE 6. — Isolation of an 18-kDa band by SECIS affinity chromatography. Sequential metal and SECIS affinity chromatography of aqueous extracts of the wild-type strain N2 (as control) and the transgenic strain IH11, ciuEx3[pLO109(Psur-5::scbp-2::polyHis); pRF4(rol-6 (su1006))] overexpressing sbp2. An 18-kDa protein band is purified from the IH11 extract (arrows), but not from the control. This mass corresponds to the expected to SBP2 if translation starts from the proposed AUU codon. (Lane 1) Molecular weight marker (MWM). (Lanes 3,4) Eluates from the Ni⁺² column. (Lanes 6,7) Flow through the wtSECIS element column. (Lanes 9,10) Eluates from the wtSECIS element column from wild type and ciuEx3, respectively (containing soluble mutSECIS). One asterisk shows the streptavidin monomer and two asterisks show the streptavidin dimer.