Construction of an Oscillator Gene Circuit by Negative and Positive Feedbacks
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Synthetic oscillators are gene circuits in which the protein expression will change over time. The delay of transcription, translation, and protein folding is used to form this kind of behavior. Here, we tried to design a synthetic oscillator by a negative feedback combined with a positive feedback. With the mutant promoter PLacC repressed by LacIq and PLux activated by AHL-bound LuxR, two gene circuits, Os-LAA and Os-ASV, were constructed and introduced into LacI-deleted E. coli DH5α cells. When glucose was used as the carbon source, a low level of fluorescence was detected in the culture, and the bacteria with Os-ASV showed no oscillation, whereas a small portion of those carrying Os-LAA demonstrated oscillation behavior with a period of about 68.3 ± 20 min. When glycerol was used as the carbon source, bacteria with Os-ASV demonstrated high fluorescence value and oscillation behavior with the period of about 121 ± 21 min.
Abstract
Synthetic oscillators are gene circuits in which the protein expression will change over time. The delay of transcription, translation, and protein folding is used to form this kind of behavior. Here, we tried to design a synthetic oscillator by a negative feedback combined with a positive feedback. With the mutant promoter PLacC repressed by LacIq and PLux activated by AHL-bound LuxR, two gene circuits, Os-LAA and Os-ASV, were constructed and introduced into LacI-deleted E. coli DH5α cells. When glucose was used as the carbon source, a low level of fluorescence was detected in the culture, and the bacteria with Os-ASV showed no oscillation, whereas a small portion of those carrying Os-LAA demonstrated oscillation behavior with a period of about 68.3 ± 20 min. When glycerol was used as the carbon source, bacteria with Os-ASV demonstrated high fluorescence value and oscillation behavior with the period of about 121 ± 21 min.
참고문헌 (21)
- Andersen JB, "New unstable variants of green fluorescent protein for studies of transient gene expression in bacteria", Appl. Environ. Microbiol, 64, pp.2240~2246 (1998)
- Andrianantoandro E, "Synthetic biology: new engineering rules for an emerging discipline", Mol. Syst. Biol, 2, pp.1~14 (2006)
- Becskei A, "Engineering stability in gene networks by autoregulation", Nature, 405, pp.590~593 (2000)
- Chen L, "Modeling Biomolecular Networks in Cells: Structure and Dynamics", pp.217~232 (2010), Springer
- Danino T, "A synchronized quorum of genetic clocks", Nature, 463, pp.326~330 (2010)
- Egland KA, "Quorum sensing in Vibrio fischeri: elements of the luxI promoter", Mol. Microbiol, 31, pp.1197~1204 (1999)
- Elowitz MB, "A synthetic oscillatory network of transcriptional regulators", Nature, 403, pp.335~338 (2000)
- Friedland AE, "Synthetic gene networks that count", Science, 324, pp.1199~1202 (2009)
- Fung E, "A synthetic gene-metabolic oscillator", Nature, 435, pp.112~118 (2005)
- Fuqua WC, "Quorum sensing in bacteria: the LuxR-LuxI family of cell densityresponsive transcriptional regulators", J. Bacteriol, 176, pp.269~275 (1994)
- Garcia-Ojalvo J, "Modeling a synthetic multicellular clock: repressilators coupled by quorum sensing", Proc. Natl. Acad. Sci. USA, 101, pp.10955~10960 (2004)
- Gardner TS, "Construction of a genetic toggle switch in Escherichia coli", Nature, 403, pp.339~342 (2000)
- Goodwin BC, "Oscillatory behavior in enzymatic control processes", Adv. Enzyme Regul, 3, pp.425~428 (1965)
- Hu FX, "Approaches and strategies of gene scarless knockout in the Escherichia coli genome", Lett. Biotechnol, 4, pp.552~557 (2013)
- Jobe A, "lac Repressor-operator interaction: IX. The binding of lac repressor to operators containing Oc mutations", J. Mol. Biol, 85, pp.231~248 (1974)
- Kaznessis YN, "Models for synthetic biology", BMC Syst. Biol, 1, pp.1~47 (2007)
- Keiler KC, "Role of a peptide tagging system in degradation of proteins synthesized from damaged messenger RNA", Science, 271, pp.990~993 (1996)
- Lutz R, "Independent and t ight regulation of transcriptional units in Escherichia coli via the LacR", O, the TetR, I1-I2 regulatory elements, pp.Nucleic Acids Res, 1997[DOI]
- O’Briena EL, "Modeling synthetic gene oscillators", Math. Biosci, 236, pp.1~15 (2012)
- Sayuta DJ, "Slow activator degradation reduces the robustness of a coupled feedback loop oscillator", Mol. Biosyst, 6, pp.1469~1474 (2010)
- Song K, "Introduction to Synthetic Biology", pp.65~83 (2012), Science Press