By Luc Ponchon
ISBN-10: 1493927299
ISBN-13: 9781493927296
This quantity offers options on contemporary advancements that use RNA scaffolds as molecular instruments. RNA Scaffolds: tools and Protocols courses readers via tools on a variety of domain names as molecular biology, nanotechnologies, and structural biology. Written for the Methods in Molecular Biology sequence, chapters contain introductions to their respective subject matters, lists of the required fabrics and reagents, step by step, without difficulty reproducible laboratory protocols and pointers on troubleshooting and warding off recognized pitfalls.
Practical and authoritative, RNA Scaffolds: tools and Protocols will function a useful reference for these attracted to extra examine into this interesting field.
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Extra info for RNA Scaffolds: Methods and Protocols
Sample text
This is possible because tRNAs are recognized and processed by a number of cellular enzymes. Furthermore, as a consequence of their three-dimensional structure, tRNAs are extremely stable to both heat-unfolding and nucleases [5]. This allows them to escape degradation and accumulate in E. coli when overexpressed from a recombinant plasmid. We took advantage of these specific features to express recombinant RNA as chimeras, by inserting them into the protective scaffold of a tRNA [6, 7]. We developed a new strategy to perform RNA–protein coexpression using E.
Ponchon L, Dardel F (2007) Recombinant RNA technology: the tRNA scaffold. Nat Methods 4:571–576 7. Ponchon L, Beauvais G, Nonin-Lecomte S, Dardel F (2009) A generic protocol for the 31 expression and purification of recombinant RNA in Escherichia coli using a tRNA scaffold. Nat Protoc 4:947–959 8. Pasloske BL, Walkerpeach CR, Obermoeller RD, Winkler M, DuBois DB (1998) Armored RNA technology for production of ribonucleaseresistant viral RNA controls and standards. J Clin Microbiol 36:3590–3594 9.
Structural alignment of untreated and optimally treated crystals reveal that the ternary complexes of T-box-tRNA-YbxF shift closer to each other in the crystal as rigid bodies (Fig. 3a; see Note 3), producing superior packing contacts such as three intimate base-stacking interactions between symmetry-related complexes (Fig. 3b) as well as a stable A-minor interaction between the engineered GAAA tetraloop on tRNA acceptor stem and the minor groove of the proximal region of T-box Stem I (Fig. 3c).