Difference between revisions of "Part:BBa K2555003"

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<partinfo>BBa_K2555003 parameters</partinfo>
 
<partinfo>BBa_K2555003 parameters</partinfo>
 
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<h4>Method</h4>
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<p>In order to show that our synthetic bacteria have a fully functional part that works, expression measurements were made. The construct was linked to sf GFP.  For the parts plate reader  was done. A plate reader is able to take optical density (OD600nm) and fluorescent measurements over time. OD is a measure of bacterial growth over time and fluorescence is a measure of protein expression over time.</p>
 
===References===
 
===References===
 
(1)Danya J. Martell, Chandra P. Joshi, Ahmed Gaballa, Ace George Santiago, Tai-Yen Chen, Won Jung, John D. Helmann, and Peng Chen (2015) “Metalloregulator CueR biases RNA polymerase’s kinetic sampling of dead-end or open complex to repress or activate transcription” Proc Natl Acad Sci U S A. 2015 Nov 3; 112(44): 13467–13472.
 
(1)Danya J. Martell, Chandra P. Joshi, Ahmed Gaballa, Ace George Santiago, Tai-Yen Chen, Won Jung, John D. Helmann, and Peng Chen (2015) “Metalloregulator CueR biases RNA polymerase’s kinetic sampling of dead-end or open complex to repress or activate transcription” Proc Natl Acad Sci U S A. 2015 Nov 3; 112(44): 13467–13472.

Revision as of 00:58, 10 October 2018


pBAD-RBS-sfGFP

BBa_K2555003 contains pBAD promoter, RBS, and superfolder GFP. The construct is used to indirectly reflect the expression of cueR of BBa_K2555002 under different concentration of L-arabinose by measuring green fluorescent intensity with plate reader. BBa_K2555002 contains pBAD (L-arabinose inducing promoter) with RBS and CueR ( a transcription factor that can bind on pcopA of BBa_K2555000). CueR behaves as a net activator or a net repressor under different copper concentrations(1). BBa_K2555002 and BBa_K2555000 are used for construction of BBa_K2555004.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1311
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1250
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1085
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 1067
    Illegal SapI.rc site found at 1355


Method

In order to show that our synthetic bacteria have a fully functional part that works, expression measurements were made. The construct was linked to sf GFP. For the parts plate reader was done. A plate reader is able to take optical density (OD600nm) and fluorescent measurements over time. OD is a measure of bacterial growth over time and fluorescence is a measure of protein expression over time.

References

(1)Danya J. Martell, Chandra P. Joshi, Ahmed Gaballa, Ace George Santiago, Tai-Yen Chen, Won Jung, John D. Helmann, and Peng Chen (2015) “Metalloregulator CueR biases RNA polymerase’s kinetic sampling of dead-end or open complex to repress or activate transcription” Proc Natl Acad Sci U S A. 2015 Nov 3; 112(44): 13467–13472.