Difference between revisions of "Part:BBa K2467000"

Revision as of 12:04, 1 November 2017

DsbA DspB emGFP

To improve part BBa_K1659211, DsbADspB (dispersin exported to the extracellular using the DsbA signal sequence (KKIWLALAGLVLAFSASA)), we fused it to emGFP in order to measure secreted protein using fluorescence. We amplified the DsbADspB fragment and incorporated EcoR1 and BamH1 restriction sites. The gene was ligated onto the multiple cloning site on the pRSET emGFP vector which resulted in fluorescently tagged DspADspB. This fragment will then be moved to pSB1C3. This allows for easy fluorescence detection in the supernatant contributed by the exported DspADspB. Projects dealing with disruption of Biofilms can easily follow the release of the dispersin enzyme into the extracellular environment to degrade the biofilm polysaccharides

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
INCOMPATIBLE WITH RFC[25]
Illegal AgeI site found at 316
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI site found at 472

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K2467000 short</partinfo>
-For our improvement of part BBa_K1659211 which is DspADspB (dispersin exported to the extracellular), we amplified the DspADspB and incorporated EcoR1 and BamH1 restriction sites. The gene was ligated onto the multiple cloning site on the pRSET emgfp vector which resulted in fluorescently tagged DspADspB. This allows for easy fluorescence detection in the supernatant contributed by the exported DspADspB.
+To improve part BBa_K1659211, DsbADspB (dispersin exported to the extracellular using the DsbA signal sequence (KKIWLALAGLVLAFSASA)), we fused it to emGFP in order to measure secreted protein using fluorescence. We amplified the DsbADspB fragment and incorporated EcoR1 and BamH1 restriction sites. The gene was ligated onto the multiple cloning site on the pRSET emGFP vector which resulted in fluorescently tagged DspADspB. This fragment will then be moved to pSB1C3. This allows for easy fluorescence detection in the supernatant contributed by the exported DspADspB.
 Projects dealing with disruption of Biofilms can easily follow the release of the dispersin enzyme into the extracellular environment to degrade the biofilm polysaccharides