Difference between revisions of "Part:BBa K833000"

Latest revision as of 03:40, 4 October 2012

Thermosensor with melting temperature of 35 to 37'C

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 plate and verified in liquid B-gal culture.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

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 __NOTOC__
 <partinfo>BBa_K833000 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.  Part BBa_K619889 is a thermosensor for Listeria monocytogenes which normally responds to a 30' to 37'c shift.   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.  We monitored thermosensor activity via the reporter LacZ.  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).
+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 plate and verified in liquid B-gal culture.
 <!-- Add more about the biology of this part here