Difference between revisions of "Part:BBa K833008"
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<partinfo>BBa_K833008 short</partinfo> | <partinfo>BBa_K833008 short</partinfo> | ||
| − | This part was derived from part BBa_K619889 via random mutagenesis and screening for thermosensors that responded to a 35 to 37'C shift. | + | |
| + | This part was derived from part BBa_K619889 via TAQ random mutagenesis and screening for thermosensors that responded to a 35 to 37'C shift. In contrast to the our wild type thermosensor (part BBa_K619889) which activates translation from a 30 to 37 degree shift, this thermosensor activates transcription from a 35 to 37 degree shift (a more concise dynamic range). This mutation was isolated by randomly mutagenizing part BBa_K619889 and monitoring a lacZ output. The thermosensor has a shine delgarno and initiating ATG that are barried within a stem-loop structure at 30'C, but are accessible at higher temperatures as the mRNA structure is disrupted. This thermosensor is part of a library of thermosensors from this screen (see parts (Bba_K833000 to Bba_K833010 and also BBa_K619890,BBa_K61989 -619894 from our 2011 submission). mutated prfA thermosensor from listeria D: This is the prfA-UTR from Listeria monocytogenes which has been randomly mutated to produce a thermosensor this a more discrete dynamic range. | ||
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| + | To use: fuse a reporter (such as LacZ, GFP or RFP) to the initiating ATG in frame | ||
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| + | Plate assay: In contrast to the our wild type thermosensor (part BBa_K619889) which activates translation from a 30 to 37 degree shift, this thermosensor activates transcription from a 35 to 37 degree shift. This phenotype has been confirmed 3 times on X-gal plates using the LacZ reporter and also in liquid B-gal assays. There is essentially no LacZ at 30'C. | ||
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Latest revision as of 04:25, 4 October 2012
Mutated prfA-UTR thermosensor from Listeria monocytogenes
This part was derived from part BBa_K619889 via TAQ random mutagenesis and screening for thermosensors that responded to a 35 to 37'C shift. In contrast to the our wild type thermosensor (part BBa_K619889) which activates translation from a 30 to 37 degree shift, this thermosensor activates transcription from a 35 to 37 degree shift (a more concise dynamic range). This mutation was isolated by randomly mutagenizing part BBa_K619889 and monitoring a lacZ output. The thermosensor has a shine delgarno and initiating ATG that are barried within a stem-loop structure at 30'C, but are accessible at higher temperatures as the mRNA structure is disrupted. This thermosensor is part of a library of thermosensors from this screen (see parts (Bba_K833000 to Bba_K833010 and also BBa_K619890,BBa_K61989 -619894 from our 2011 submission). mutated prfA thermosensor from listeria D: This is the prfA-UTR from Listeria monocytogenes which has been randomly mutated to produce a thermosensor this a more discrete dynamic range.
To use: fuse a reporter (such as LacZ, GFP or RFP) to the initiating ATG in frame
Plate assay: In contrast to the our wild type thermosensor (part BBa_K619889) which activates translation from a 30 to 37 degree shift, this thermosensor activates transcription from a 35 to 37 degree shift. This phenotype has been confirmed 3 times on X-gal plates using the LacZ reporter and also in liquid B-gal assays. There is essentially no LacZ at 30'C.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]