1. Characterization of the Hrp Regulatory System Project 1 of the Imperial iGEM 2007 team

2. Introduction In Pseudomonas syringae strains, the HrpL- dependent regulon encodes a type III protein translocation complex to translocate effector proteins required for pathogenesis. HrpR and HrpS function as positive regulatory factors for the hrpL promoter, but their mechanism of action has not been established. Structurally related to enhancer-binding proteins, but they lack receiver domains and do not appear to require a cognate protein kinase for activity Can it be characterized for Synthetic Biology?!

3. Background information and review of literature General introduction given by Imperial researchers Additional information found in literature (journals/suggested reading) Vibrant scientific community

4. Project Plan

5. Specifications Experiments will attempt to fulfil an objective within a certain context, and with variation of a single parameter for the listed components.objectivecontext parametercomponents Objective Prioritised list of experimental objectives Context Characterization in different chassis Parameters Using different parts of the Registry Components Prioritised list of parts and devices

6. Specifications Top 3 parts and devices to characterize:

7. Project Plan

8. Experimental Design Experimental design involves parts and protocols: List of Required Parts Well-defined and reliable parts from the Registry DNA constructs from GeneART Protocols Measuring PoPs Various techniques to measure gene expression

9. Measuring PoPS Polymerase Per Second (PoPS) Current of Gene expression Generic Unit DEVICE PoPS IN PoPS OUT

10. Measuring PoPS BBa_F2620 promoter used units: Molecules of GFP synthesised per cell per second. DEVICE RFP cell -1 sec -1 GFP cell -1 sec -1

11. Experimental Design Desired result of measuring input and output using fluorescent proteins

12. Project Plan

13. Timeline Modelling Using Cell Designer and other tools Estimated: Week 3-4 Implementation After wet lab intro, and upon receipt of constructs from GeneART Estimated: Week 4-8 Testing Validation Duplicate results, data analysis that corresponds with Modelling and validates the experiment Win iGEM!

15. Questions? End of Presentation