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Team:SDU-Denmark/Project
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Contents |
Overall project
Bactobandage
Inhibition of quorum-sensing mediated biofilm-formation in staphylococcus aureus.
We're going to stop Staphylococcus aureus infections (not colonization) by inhibiting the quorum sensing-dependent biofilm formation
Initial phase planning
Preliminary Project Details
The details we need to settle on soon (eg. by Thursday) are:
1) Which bacteria is the producer in the bacto bandage (suggestions: E. coli, Lactococcus, ??)
2) Identifying the natural producer of the quorum quenching signal (QQS)
3) Which proteins should be expressed
4) How should expression be regulated
5) How do we retain plasmid stability in the bactobandage
6) More?
Initial Experimental Considerations
Before we can get started a number of experimental procedures need to be discussed and/or protocols need to be identified:
Initial lab work:
1) Making competent E. Coli for our freezer
2) Amplification of basic parts in E. Coli
3) Plasmid purification (maxi prep og mini prep)
4) Growth and handling of reciever bacteria
5) Identify and amplify our favourite plasmid backbones
Biobrick expression:
1) Planning and ordering primers
2) Amplification of target gene from chromosomal DNA (PCR)
3) Manipulation of biobrick part and composite parts
4) Transformation/electroporation into producer bacteria
Testing output:
1) Western blot (protein)
2) Northern blot (RNA)
3) Promoter LaxZ or XFP fusions
4) RT-PCR
5) Sequencing (a service we buy)
6) More???
