Difference between revisions of "Part:BBa K2995000"
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To characterize our promoter we cloned it upstream of the RBS in <partinfo>BBa_E0240</partinfo> (RBS-GFP-Term) and compared it's GFP expression with the already well characterized <partinfo>BBa_J23101</partinfo> promoter in <partinfo>BBa_I20260</partinfo>. To limit the impact of other variables we insured that <partinfo>BBa_I20260</partinfo> and <partinfo>BBa_E0240</partinfo> shared the same backbone, RBS and terminator. In addition to characterizing this promoter with respect to<partinfo>BBa_I20260</partinfo> we also confirmed proper function of this promoter in the pRJPaph-bjGFP plasmid which was used for expression in Bradyrhizobium for our 2019 project.   
 
To characterize our promoter we cloned it upstream of the RBS in <partinfo>BBa_E0240</partinfo> (RBS-GFP-Term) and compared it's GFP expression with the already well characterized <partinfo>BBa_J23101</partinfo> promoter in <partinfo>BBa_I20260</partinfo>. To limit the impact of other variables we insured that <partinfo>BBa_I20260</partinfo> and <partinfo>BBa_E0240</partinfo> shared the same backbone, RBS and terminator. In addition to characterizing this promoter with respect to<partinfo>BBa_I20260</partinfo> 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
  
 
1. Cell were grown overnight in LB media at 37 degrees
 
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.
 
2. The following morning GFP expression was quantified using the Synergy 4 BioTek plate reader.
  

Revision as of 23:21, 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_I20260 and BBa_E0240 shared the same backbone, RBS and terminator. In addition to characterizing this promoter with respect toBBa_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:


(Excitation: 485, Emmission: 525)