Composite

Part:BBa_K3376002

Designed by: Li, Sheng-Fong   Group: iGEM20_Mingdao   (2020-10-06)


ldhp-GFP-Tr/pSB1C3

Lactate dehydrogenase catalyzes the fermentation of pyruvate to lactic acid. It plays a pivotal role in the glucose metabolic pathway connecting glycolysis, and is recognized as an essential gene and expressed at high level in S. mutans. Its promoter was cloned out from the gDNA of S. mutans and tested in transformed E. coli.

Expression

The fluorescence expression levels were measured for lactate dehydrogenase promoter (ldhp) [ldhp-GFP-Tr/pSB1C3 (BBa_K3376002)] and thiol peroxidase promoter (tpxp) [tpxp-GFP-Tr/pSB1C3 (BBa_K3376004)] activity of S. mutans. GFP was detected at Ex/Em = 483/513. As shown in the figure, the expression level of ldhp-driven GFP was very high compared to a regulated promoter (tpxp) and a vector-only control.

T--Mingdao--ww1.png

Transformation

The ldhp-GFP-Tr/pSB1C3 [BBa_K3376002] was transformed into E. coli Nissle strain. The high expression of green fluorescent protein was observed under a blue led light.

T--Mingdao--ww2.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
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 814


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