Template:SBB12 part sbb1216: Difference between revisions
JCAnderson (talk | contribs) (New page: <pre> Partname: sbb1216 Featurename: cI-N-GFP-B Genename: cI, GFP Source: Lambda phage / via the Registry </pre> This one is a little different. You aren't putting a homodi...) |
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<pre> | <pre> | ||
Partname: sbb1216 | Partname: sbb1216 | ||
Featurename: cI-N- | Featurename: cI-N-AmilCP-B | ||
Genename: cI, | Genename: cI, AmilCP | ||
Source: Lambda phage / via the Registry | Source: Lambda phage / via the Registry | ||
</pre> | </pre> | ||
This one is a little different. You aren't putting a homodimer onto ToxR; you're testing out a different homodimerization system. You're replacing ToxR with a different dimerization reporter, and then fusing it with amilCP, a blue protein, which will facilitate future cloning. This one is the cI transcription reprepressor from lambda phage, which has been shown to do similar reporting of homodimerization as is reported for ToxR . | This one is a little different. You aren't putting a homodimer onto ToxR; you're testing out a different homodimerization system. You're replacing ToxR with a different dimerization reporter, and then fusing it with amilCP, a blue protein, which will facilitate future cloning. This one is the cI transcription reprepressor from lambda phage, which has been shown to do similar reporting of homodimerization as is reported for ToxR PMID: 10411275. | ||
Your final construct should be a normal BglBricks part in plasmid pBca9525. You most likely will want to piece the part together using SOEing to get something like: | |||
EcoRI-BglII-Promoter-rbs.cI-spacer-NheI-spacer-rbs.AmilCP!-BamHI | |||
{{SBB12_TemplateDNAAvailable}} | {{SBB12_TemplateDNAAvailable}} | ||
Revision as of 17:41, 5 February 2012
Partname: sbb1216 Featurename: cI-N-AmilCP-B Genename: cI, AmilCP Source: Lambda phage / via the Registry
This one is a little different. You aren't putting a homodimer onto ToxR; you're testing out a different homodimerization system. You're replacing ToxR with a different dimerization reporter, and then fusing it with amilCP, a blue protein, which will facilitate future cloning. This one is the cI transcription reprepressor from lambda phage, which has been shown to do similar reporting of homodimerization as is reported for ToxR PMID: 10411275.
Your final construct should be a normal BglBricks part in plasmid pBca9525. You most likely will want to piece the part together using SOEing to get something like:
EcoRI-BglII-Promoter-rbs.cI-spacer-NheI-spacer-rbs.AmilCP!-BamHI
Genomic or plasmid DNA with this feature is available
Either genomic DNA or plasmid DNA is available for this sequence. Use your usual considerations of size and number of internal restriction sites to decide whether you should use that template for construction. You'll also need to think through which polymerase to use in making the construct.
