The spinach RNA aptamer as a characterization tool for synthetic biology
| Autor: | Francesca Ceroni, Georgios Pothoulakis, Tom Ellis, Benjamin Reeve |
|---|---|
| Přispěvatelé: | Engineering & Physical Science Research Council (E, Engineering & Physical Science Research Council (EPSRC) |
| Jazyk: | angličtina |
| Rok vydání: | 2013 |
| Předmět: |
EXPRESSION
Biochemistry & Molecular Biology Aptamer Biomedical Engineering Biology Biochemistry Genetics and Molecular Biology (miscellaneous) Ribosome Biochemical Research Methods Green fluorescent protein PROMOTERS Spinacia oleracea FLUORESCENT PROTEIN Gene expression ribosome binding site Protein biosynthesis Escherichia coli RIBOSOME Messenger RNA Science & Technology promoter Binding Sites Spinach RNA POLYMERASE RNA aptamer General Medicine Aptamers Nucleotide Molecular biology Ribosomal binding site Cell biology Luminescent Proteins RNA Bacterial ESCHERICHIA-COLI Synthetic Biology measurement fluorescence Genetic Engineering Life Sciences & Biomedicine |
| Popis: | Characterization of genetic control elements is essential for the predictable engineering of synthetic biology systems. The current standard for in vivo characterization of control elements is through the use of fluorescent reporter proteins such as green fluorescent protein (GFP). Gene expression, however, involves not only protein production but also the production of mRNA. Here, we present the use of the Spinach aptamer sequence, an RNA mimic of GFP, as a tool to characterize mRNA expression in Escherichia coli. We show how the aptamer can be incorporated into gene expression cassettes and how co-expressing it with a red fluorescent protein (mRFP1) allows, for the first time, simultaneous measurement of mRNA and protein levels from engineered constructs. Using flow cytometry, we apply this tool here to evaluate ribosome binding site sequences and promoters and use it to highlight the differences in the temporal behavior of transcription and translation. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|