Difference between revisions of "Part:BBa K1962006"
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===Usage and Biology=== | ===Usage and Biology=== | ||
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| + | {|width='80%' style='border:1px solid gray' | ||
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| + | <partinfo>BBa_K892008 AddReview 5</partinfo> | ||
| + | <I>Parts Collection 2016</I> | ||
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| + | This is part of a Part Collection of 17 BioBricks designed by Dundee iGEM 2016. This collection will be useful to teams working with toxins as we have submitted new toxins to the registry. Working with bacterial toxins is difficult due to the risk of toxicity to the chassis, so the corresponding immunity for our toxins were also submitted. We have also submitted these toxins lacking their cytotoxic domains replacing it with a multiple cloning site which will allow for different toxic domains to be fused at the C-terminus and thereby generating a synthetic toxin. In addition, there are three well-characterised promoters that can be used to initiate gene expression at various points in the digestive tract, to enable devices to function within a human or animal. Finally, a lysis cassette was constructed to lyse or burst cells, thus releasing the toxins and destroying the GM bacteria to prevent its release to the environment. | ||
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| + | This <partinfo>BBa_K1962005</partinfo> is an immunity protein against colicin E9, which is not currently in The Registry. This part will be useful for teams wishing to clone colicin E9 in the future, helping to neutralise the toxin during cloning of the gene | ||
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Revision as of 23:33, 29 October 2016
Truncated Colicin E9 Lacking Bacteriocin Active Domain
Colicins are anti-bacterial proteins produced by some strains of E. coli that typically have three domains: a translocation domain; a receptor binding domain; and a cytotoxic domain. This biobrick encodes a truncated version of Colicin E9 which lacks the C-terminal bacterocin domain, which is this case is a DNase.
The part has an engineered multiple cloning site at the 3' end, preceding the RFC[10] suffix and regulation double stop codons, containing the following in-frame restriction sites: BamHI, KpnI, SalI, BglII and NheI. The presence of this multiple cloning site will allow for different toxic domains, or other polypeptides, to be fused at the C-terminus of the truncated colicin and thereby generating a synthetic colicin or novel fusion protein in a rapid and facile manner.
This Biobrick was used to generate two synthetic colicins (BBa_K1962007 and BBa_K1962008).
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 1381
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1375
Illegal BamHI site found at 1357 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]