Difference between revisions of "Part:BBa K2995000"
(→‎Characterization)
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(Waterloo iGEM 2019)  
 
(Waterloo iGEM 2019)  
  
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To characterize our promoter we cloned it upstream of the RBS in BBa_E0240 (RBS-GFP-Term) and compared it's GFP expression with the already well characterized BBa_J23101 promoter in BBa_I20260. To limit the impact of other variables we insured that BBa_J23101 and BBa_E0240 shared the same backbone, RBS and terminator. In addition to characterizing this promoter with respect to BBa_I20260 we also confirmed proper function of this promoter in the  
+
To characterize our promoter we cloned it upstream of the RBS in BBa_E0240 (RBS-GFP-Term) and compared it's GFP expression with the already well characterized BBa_J23101 promoter in BBa_I20260. To limit the impact of other variables we insured that BBa_J23101 and BBa_E0240 shared the same backbone, RBS and terminator. In addition to characterizing this promoter with respect to BBa_I20260 we also confirmed proper function of this promoter in the pRJPaph-bjGFP plasmid which was used for expression in Bradyrhizobium for our 2019 project. 
  
 
Note: empty DH5alpha was used as a negative control for GFP expression
 
Note: empty DH5alpha was used as a negative control for GFP expression
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2. The following morning GFP expression was quantified using the Synergy 4 BioTek plate reader.
 
2. The following morning GFP expression was quantified using the Synergy 4 BioTek plate reader.
  
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The graph below indicates the results from this characterization exper
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The graph below indicates the results from this characterization experiment:

Revision as of 23:13, 21 October 2019


Paph promoter

Constitutive promoter for expression in E. coli and B. japonicum.

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]


Characterization

(Waterloo iGEM 2019)

To characterize our promoter we cloned it upstream of the RBS in BBa_E0240 (RBS-GFP-Term) and compared it's GFP expression with the already well characterized BBa_J23101 promoter in BBa_I20260. To limit the impact of other variables we insured that BBa_J23101 and BBa_E0240 shared the same backbone, RBS and terminator. In addition to characterizing this promoter with respect to BBa_I20260 we also confirmed proper function of this promoter in the pRJPaph-bjGFP plasmid which was used for expression in Bradyrhizobium for our 2019 project.

Note: empty DH5alpha was used as a negative control for GFP expression

1. Cell were grown overnight in LB media at 37 degrees 2. The following morning GFP expression was quantified using the Synergy 4 BioTek plate reader.

The graph below indicates the results from this characterization experiment: