Towards Intelligent Probiotics. Chris Brasseaux , EE David Golynskiy , Bio/ Crim Tyler Guinn, BioChem /EE Sameer Sant , Bio/Econ Mitu Bhattatiry , Biomed (Columbia) Nimi Bhattatiry (High School) Jose Alfredo Flores (Monterey Mexico). Towards Intelligent Probiotics. Introduction
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Chris Brasseaux, EE
Tyler Guinn, BioChem/EE
NimiBhattatiry (High School)
Jose Alfredo Flores (Monterey Mexico)
Image from Kolmer, 1994
1. Transformed E.Coli Dh5a
2. Incubated with nitrates for production of the receptor
3. Introduced heparin and FGF
4. Performed fluorescent microscopy measurements
Image from Roland Institute at Harvard
Image from Roland McGraw Hill
The signaling network from the input of external ligand signal to the output of the tumbling state of a E coli cell can be quantitatively described by a modular model.
The model is formulated based on the law of mass action and Michaelis-Menten mechanism and contains four relatively independent modules.Module 1: Activation of ToxR receptor
Module 2: Transcription/translation of CheZModule 3:CheYdephosphorylation by the CheZ proteinModule 4: The tumbling activity of E coli is characterized by the so-called “bias”, which is defined as the ratio of the time of directed movement and the total time. It is experimentally measured that the bias is a Hill function dependent on the concentration of phosphorylatedCheY (Cluzel, Surette et al. 2000).
Population 1 (Immunobot)
Population 1 (Immunobot) + Population 2 (Killbot)
Population 1 (Immunobot) + more Population 2 (Killbot)
The killbots eliminate the majority of the cells
The addition of glucose in the medium increases the population two-fold
Built new BioBricks relating to wound sensing and chemotactic abilities
Demonstrated that some of them, particularly the killbot, seem to work as expected.
Improved the characterization existing BioBricks, and included our experience in the appropriate Registry page.
Qualifying for MIT will allow us to test the different promoter-receptor constructs with longer induction times, different ligand concentrations, and the chemotaxis experiments with controlled gradient settings.
Dr. Hyun-Joo Nam
Dr. Lan Ma