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. 2018 Dec 11;9(3):639–647. doi: 10.1016/j.apsb.2018.12.002

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© 2019 Chinese Pharmaceutical Association and Institute of Materia Medica, Chinese Academy of Medical Sciences. Production and hosting by Elsevier B.V.

This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

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Fig. 1 — Bioengineering of new BERA/miR-27b-3p molecule. (A) Schematic illustration of the tRNA/pre-miR-34a carrier used for the production of BERA/miR-27b-3p. (B) Urea-PAGE analysis of total bacterial RNAs showed that chimeric miR-27b-3p was heterogeneously expressed in E. coli using the tRNA/pre-miR-34a scaffold. Total RNAs isolated from untransformed HST08 (WT) E. coli were used as control, and our previously constructed tRNA-hsa-miR-27b-123nt plasmid¹⁹ transformed HST08 E. coli was utilized for comparison. MiR-27b-3p was expressed at much higher levels when using the tRNA/pre-miR-34a carrier. (C) Representative FPLC traces during the purification of BERA/miR-27b-3p. Inserts were corresponding urea-PAGE analyses of collected fractions (1, 2, 3 and 4) eluted at 8.7 min, which confirmed the purity of isolated recombinant ncRNAs.