Team:Warwick/Parts
Parts
Our new parts are all versions of the same modular system, a fusion protein formed from a bacterial transmembrane domain and a zinc-finger domain, with linker domains between domains and a FLAG-tag for immunofluorescence staining. Each transmembrane/zinc-finger domain is independently interchangeable with other transmembrane/zinc-finger domains, though not interchangeable between each other (i.e. the transmembrane domain cannot be swapped for a zinc-finger domain and vice versa).
The purpose of this fusion protein is to allow bacteria (specifically Escherichia coli) to bind specific short double stranded DNA sequences at their surface; using immobilised DNA the cell should remain bound in place, or this protein can be used for cell identification (barcoding).
The purpose of this fusion protein is to allow bacteria (specifically Escherichia coli) to bind specific short double stranded DNA sequences at their surface; using immobilised DNA the cell should remain bound in place, or this protein can be used for cell identification (barcoding).
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Lpp-OmpA-Zif268
This part is our construct with Lpp-OmpA as the transmembrane domain and the Zif268 1AAY fragment as the zinc-finger domain. The transmembrane domain can be exchanged with another using NdeI and AgeI, and the zinc-finger domain can be exchanged using NheI and ClaI. The entire construct is under the pL_lac promoter, and contains a C-terminal FLAG tag.
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Lpp-OmpA-sZF2
This part is our construct with Lpp-OmpA as the transmembrane domain and sZF2 as the zinc-finger domain. The transmembrane domain can be exchanged with another using NdeI and AgeI, and the zinc-finger domain can be exchanged using NheI and ClaI. The entire construct is under the pL_lac promoter, and contains a C-terminal FLAG tag.
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Lpp-OmpA-sZF10
This part is our construct with Lpp-OmpA as the transmembrane domain and sZF10 as the zinc-finger domain. The transmembrane domain can be exchanged with another using NdeI and AgeI, and the zinc-finger domain can be exchanged using NheI and ClaI. The entire construct is under the pL_lac promoter, and contains a C-terminal FLAG tag.
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Lpp-OmpA-sZF14
This part is our construct with Lpp-OmpA as the transmembrane domain and sZF14 as the zinc-finger domain. The transmembrane domain can be exchanged with another using NdeI and AgeI, and the zinc-finger domain can be exchanged using NheI and ClaI. The entire construct is under the pL_lac promoter, and contains a C-terminal FLAG tag.
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INP-Zif268
This part is our construct with INP as the transmembrane domain and the Zif268 1AAY fragment as the zinc-finger domain. The transmembrane domain can be exchanged with another using NdeI and AgeI, and the zinc-finger domain can be exchanged using NheI and ClaI. The entire construct is under the pL_lac promoter, and contains a C-terminal FLAG tag.
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BclA-Zif268
This part is our construct with BclA as the transmembrane domain and the Zif268 1AAY fragment as the zinc-finger domain. The transmembrane domain can be exchanged with another using NdeI and AgeI, and the zinc-finger domain can be exchanged using NheI and ClaI. The entire construct is under the pL_lac promoter, and contains a C-terminal FLAG tag.