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Header
Cell Surface DisplayInverted Microscopy
- Inoculation overnight of all our constructs (ISA,ISD,ISI and competent cells as negative control) to do microscopy.
Sensing-Effector
PCR of hya-construct (sensing), MMP2-construct (effector) and MMP9-construct (effector)
-After the Minipreps of the innocuated bacteria I did a 20ul PCR reaction in order to check if the Bacteria did indeed contain my plasmids using primers specific only to the insert. The PCR results showed that the hya-constructs and the MMP2-construct were correct but that the construct with MMP9 did not work. These conclusions were also supported by the Microscopy we did for the constructs MMP2 and MMP9. The Bacteria with the MMP9 construct were bright green which they should not have been since instead of GFP they should express MMP9 whereas the MMP2 Bacteria did not express GFP.
Inoculation of the constitutive promoter construct and the GelE construct containing Bacteria
-These two constructs were the two last ones that I had not yet verified by PCR. So I inoculated them in order to do a MiniPrep and a Test PCR.
Note
Because time was running short, we decided to characterize only the following constructs:
1.) HyaPromoter+GFP
2.) CadPromoter+GFP
3.) ConstitutivePromoter+GFP
4.) AraCPromoter+GelE
5.) AraCPromoter+MMP2
Nanoparticles
Making/loading Nanoparticles
-Made rGFP loaded nanoparticles.
-Made FITC-dextran nanoparticles.
