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Advanced Predictive Modelling of Meat Spoilage Using Cell-Free Systems

This presentation outlines a cutting-edge project aimed at predicting meat spoilage levels through the use of cell-free systems. It covers background information, project approaches, and progress made, including collaborations between lab, modeling, and protocols teams. Key objectives include testing the effects of temperature on gene expression, generating predictive models, and characterizing specific constructs. The presentation will conclude with a Q&A session to discuss ongoing phases and next steps, ensuring the development of efficient meat spoilage prediction mechanisms.

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Advanced Predictive Modelling of Meat Spoilage Using Cell-Free Systems

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Presentation Transcript

  1. CELL BY DATE UPDATE 29.08.07 14:00

  2. Presentation Outline • CBD Background • Project Approach • What’s Done • Lab Team • Modelling Team • Protocols Team • What’s Next • Q & A

  3. Background Background Project Approach What’s Done What’s Next Q & A • Specifications • Reports Thermal Exposure • Accurately predict level of meat spoilage • Use of cell-free systems • CBD Reporter System • Effects of temperature on gene expression • Constitutive promoter • Fluorescent protein (Example of a CBD DNA Construct)

  4. Project Approach Background Project Approach What’s Done What’s Next Q & A Phase 1 Phase 2 Phase 3 Aim: Initial Testing • Build DNA constructs • Initial Testing in vivo • Initial Testing in vitro • Generate equations for Modelling Aim: Characterization of Specific Construct • Operating Range • Effects of Temp. on • Fluorescence vs. Time • Lifespan • Generate preliminary data for Modelling Aim: Testing/Validation of Predictive Model • Accurate prediction of outcome • Fluorescence vs. Time • Lifespan • Corroborate specs of device • DsRed-Express & in-veso

  5. Project Approach: Breakdown Background Design Phases What’s Done What’s Next Q & A Lab Team Modelling Team Protocols Team Build / amplify DNA constructs Derive equations / Carry out simulations Gather / mergeprotocols Conduct Expts / Generate data for analysis Generation of predictive model

  6. What’s Done Background Project Approach What’s Done What’s Next Q & A • Lab Team • Cloning/Amplification • pTet-GFP • pT7-GFP • Biobricks construct does not work well • To be obtained from different sources • pCI-GFP (Cancelled) • Awaiting DsRed-Express

  7. What’s Done Background Project Approach What’s Done What’s Next Q & A • Modelling Team • kFP : Function of Temperature. k is based on the promoter used as promoters take time to turn on. • dFP : Function of System. May be considered to be a function of temperature as proteins degrade faster at higher temperatures.

  8. What’s Done Background Project Approach What’s Done What’s Next Q & A • Modelling Team

  9. What’s Done Background Project Approach What’s Done What’s Next Q & A • Protocols Team • Works in vivo & in vitro • Tested @ 10oC, 25oC, 37oC, 45oC • Does not work at 10oC, 45oC • But when 10oC left o/n at 25oC, gene expression occurs • Worked weakly in vivo & in vitro • Very weak output cf. pTet • To be obtained from different sources and tested

  10. What’s Done Background Project Approach What’s Done What’s Next Q & A • Protocols Team (Gaps in data due to lab restrictions) (1 magnitude increase)

  11. What’s Done Background Project Approach What’s Done What’s Next Q & A • Protocols Team (Not working well) (Difference in expression in vivo)

  12. What’s Done Background Project Approach What’s Done What’s Next Q & A Phase 1 Phase 2 Phase 3 Aim: Initial Testing • Build DNA constructs • Initial Testing in vivo • Initial Testing in vitro • Generate Equations for Modelling Aim: Characterization of Specific Construct • Operating Range • Effects of Temp. on • Fluorescence vs. Time • Lifespan • Generate preliminary data for Modelling Aim: Testing/Validation of Predictive Model • Accurate prediction of outcome • Fluorescence vs. Time • Lifespan • Corroborate specs of device • DsRed-Express & in-veso

  13. What’s Next Background Project Approach What’s Done What’s Next Q & A Timeline • Phase 2 • [GFP] & δ[GFP] curves • Homemade cell extract • Phase 3 • Suitable temp. step increases • Refine predictive model Week 8 Week 10 Week 9 • Phase 4? • Proper documentation • Substitute GFP w/ DsRed-Express • In-veso gene expression • Packaging Methods • Phase 2 • Effects of temperature • Gentle / steep gradients • Preliminary data analysis

  14. Q & A Background Project Approach What’s Done What’s Next Q & A

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