Difference between revisions of "Part:BBa K1084009"
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Theoretical promoter modeling also has done (see promoter Background page in HokkaidoU 2013 wiki).
 
Theoretical promoter modeling also has done (see promoter Background page in HokkaidoU 2013 wiki).
 
According to protein, these promoters show relatively another protein activity revel.
 
According to protein, these promoters show relatively another protein activity revel.
−
Choose your best promoter by promoter optimization kit (POK).
+
Choose your best promoter by promoter optimization kit (Promoter Selector).
−
Plate image below is actual worked POK (Km resistance).
+
Plate image below is actual worked Promoter Selector (Km resistance).
  
 
https://static.igem.org/mediawiki/parts/0/00/HokkaidoU_2013_POK_DEMO_48h_1.png
 
https://static.igem.org/mediawiki/parts/0/00/HokkaidoU_2013_POK_DEMO_48h_1.png

Revision as of 10:40, 19 October 2013

promoter 9

iGEM HokkaidoU Japan 2013 original promoter family is constructed from consensus promoter (BBa_K1084001) by adding mutation at -35 region. Other region is same among family members, and similar with promoter consensus sequence.

HokkaidoU2013_promoter_Result-fig1.png

Fig. 1 Randomized promoter family sequences

Correspondence of sequence and parts number is below. 

  -35        BBa_         t. e. rank
  TTGACA     K1084001     1   
  TAGGTC     K1084002     2
  CTGAAG     K1084003     6
  GGGGTG     K1084004     3
  GAGGAT     K1084005     5
  GCAATA     K1084006     7
  GGGGGG     K1084007     8
  TGTGTG     K1084008     4
  AGTGGG     K1084009     9
  TCTCGG     K1084010     10

t. e. = theoretical transcription efficyency

Promoter transcription efficiency was measured with mRFP1 (BBa_K1084401-K1084410), LacZ and Kanamycine (Km) resistance (BBa_K1084501-K1084505). HokkaidoU2013_promoter_Result-fig2.png

Fig. 2 mRFP1 assay result

HokkaidoU2013_promoter_Result-fig4.png

Fig. 3 β-Galactosidase assay result

HokkaidoU_2013_Result-f6_1_1.png

Fig. 4 Km resistance assay result

HokkaidoU_2013_Result-f7.png

Fig. 5 Comparison of assay results and modeling data

Theoretical promoter modeling also has done (see promoter Background page in HokkaidoU 2013 wiki). According to protein, these promoters show relatively another protein activity revel. Choose your best promoter by promoter optimization kit (Promoter Selector). Plate image below is actual worked Promoter Selector (Km resistance).

HokkaidoU_2013_POK_DEMO_48h_1.png


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]