期刊
ACS SYNTHETIC BIOLOGY
卷 3, 期 3, 页码 182-187出版社
AMER CHEMICAL SOC
DOI: 10.1021/sb400089c
关键词
measurement; Spinach RNA; aptamer; promoter; ribosome binding site; fluorescence
资金
- TMO Renewables Ltd.
- U.K. Engineering and Physical Sciences Research Council (EPSRC)
- Engineering and Physical Sciences Research Council [EP/J021849/1, EP/J02175X/1] Funding Source: researchfish
- EPSRC [EP/J02175X/1, EP/J021849/1] Funding Source: UKRI
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 to highlight the differences in the temporal behavior of transcription and translation.
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