IGEM:Imperial/2010/XylE team

XylE team Lab Objectives

construction of XylE fusion protein
Testing expression of XylE in E.coli and characterization under the control of a constitutive promoter
Construction of the -ComE promoter/XylE fussion protein- expression system
Construction of the -LacI promoter/XylE fusion construct- expression system

Week 7	Monday	Tuesday	Wednesday	Thursday	Friday
MORNING	3A assemply: ligation of RBS-XyL, J23101 promoter and PSB1C3 vector (overnight)	*estimated date of primers delivery 3A assemply: transformation of E.coli with XylE construct (overnight)	3A assemply: make replica plates (overnight) Catechol assay of E.coli	Mini-prep XylE E.coli	Midi-prep XylE E.coli
lunch break
AFTERNOON		Starting construction of XylE fusion protein

Week 6	Thursday	Friday
MORNING		restriction digestion of the biobrick vector (J33204)containing the XylE gene (90min) Digest PSB1C3 vector to prepare it for 3A assemply
lunch break
AFTERNOON	Starting of “Testing expression of XylE in E.coli” objective 1)Annealing EcoRI and speI oligos to J23101 promoter which will be annealed later in front of the RBS-XylE registry gene (overnight	3) gel purification of the of the XylE gene (2hrs)