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Team:HKUST/future work
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Revision as of 19:50, 23 October 2010
Future Work for iGEM 2010 HKUST team
- Transform Lactobacillus plantarum WCFS1 with our existing constructs [agrC-mCherry/agrC-plnB-mCherry in pMG36e]. Use fluorescence microscope to examine whether the chimeric AIP receptor localizes on Lactobacillus cell membrane.
- We will continue our "chimeric AIP receptor functionality testing" experiments in two Lactobacillus species, L. plantarum WCFS1 and L. sakei Lb790. We have already built the part [agrC/agrC-plnB in pMG36e] of our original design and we hope to complete the rest [plnA promoter – gusA report unit] soon. After obtaining the final construct [plnA promoter – gusA reporter – P32 promoter – agrC/agrC-plnB], we will transform the pMG36e shuttle vector into respective Lactobacillus cells. Finally, we will apply gusA reporter assay to determine whether the fusion AIP receptor functions as expected.
- Migrate the existing constructs "RIP synthesis and secretion cassette" [SP+DD13-RIP] from pBluescript to shuttle vector pMG36e.
- Perform functionality test to examine RIP secretion efficiency in L. plantarum.
- Conduct bio-assay of the reporter plasmid and estimate the inhibition efficiency of DD13-RIP against S. aureus growth.
- Combine constructs from two modules – "AIP Sensor on Lactobacillus Cell Membrane" and "RIP Synthesis and Secretion in Lactobacillus" – together. We will then transform the combined construct [plnA promoter – RIP synthesis and secretion cassette – P32 promoter – agrC/agrC-plnB] into L. plantarum and examine if our initial design works as expected.
