This new volume of Methods in Enzymology continues the legacy of this premier serial with quality chapters authored by leaders in the field. This volume covers research methods in biomineralization science and contains sections on such topics as constructing and optimizing artificial riboswitches, live cell imaging and intracellular sensors with artificial riboswitches, conditional control of gene expression with artificial riboswitches, using artificial riboswitches for protein evolution and pathway optimization, and anti-riboswitch drug screens. Continues the legacy of this premier serial with quality chapters authored by leaders in the fieldCovers research methods in biomineralization scienceContains sections on such topics as constructing and optimizing artificial riboswitches, synthetic biology: live cell imaging and intracellular sensors with artificial riboswitches, synthetic biology: conditional control of gene expression with artificial riboswitches, synthetic biology: using artificial riboswitches for protein evolution and pathway optimization, anti-riboswitches drug screens INDICE: 1. In silico rational design of synthetic riboswitches that regulate gene expression at the transcriptional level 2. Ligand-dependent exponential amplification of self-replicating RNA enzymes3. Designing synthetic riboswitches from biological parts 4. Integrating and amplifying signals from riboswitch biosensors5. Aptazyme optimization in a cell-free transcription-translation system6. Engineering artificial riboswitches for gene regulation in yeast and non-model parasitic organisms7. Engineering of shunting-modulating eukaryotic riboswitches by using a cell-free translation system8. Constructing genetically encoded sensors by transcriptionally fusing them to fluorescent reporter genes and examining them using fluorescence microscopy9. Using Spinach2 aptamers for live cell imaging of intracellular RNA 10. Live cell imaging of intracellular cyclic dinucleotide levels with ligand-dependent Spinach aptamers.11. Using Spinach aptamer to correlate mRNA and protein levels in E coli12. Monitoring mRNA and protein levels in bulk and in model vesicle-based artificial cells13. Sequence-modulated Spinach aptamer as a reporter for in vitro nucleic acid circuits and also as a real-time reporter of isothermal nucleic acid amplification reactions for target detection and distinction of single nucleotide polymorphism14. Using riboswitches to define gene function in intracellular pathogens (Mycobacteria) through artificially regulated gene expression15. Controlling expression of essential chloroplast genes in the unicellular alga Chlamydomonas reinhardtii vitamin-repressible riboswitch.16. Conditional control of gene expression by artificial riboelements in Streptomyces coelicolor17. Using in vivo screens to identify ligand-responsive RNA switches to enable thiamine-dependent regulation of large RNAs 18. Designing riboswitches and aptazymes for engineering metabolic pathways19. Protein Engineering by coupling enzyme output (ligand) to aptazyme-regulated GFP signal20. Using riboswitches for Synthetic Biology pathway optimization (L-lysine-producing Escherichia coli)21. Fluorescence assays for monitoring RNA-ligand interactions and riboswitch-targeted drug discovery screening22. Monitoring ribosomal frameshifting as a platform to screen anti-riboswitch drug candidates
- ISBN: 978-0-12-801123-2
- Editorial: Academic Press
- Encuadernacion: Cartoné
- Páginas: 514
- Fecha Publicación: 15/01/2015
- Nº Volúmenes: 1
- Idioma: Inglés