Composite

Part:BBa_K3889150

Designed by: Prameya Garge, Ashwin Sharma   Group: iGEM21_IISER-Tirupati_India   (2021-10-06)


SP200+SRTF1 Binding Site+ RBS

SP200 (BBa_K3889012) fused with SRTF1 binding site (BBa_K3889030) and RBS (BBa_K1351031) in order to create a progesterone sensitive promoter if SRTF1 (BBa_K3889021) is being expressed.

In order to produce P22 c2 repressor (BBa_K3889020) in synchronisation with progesterone this part was used as in BBa_K3889101.

Results

T--IISER-Tirupati India--Progesterone sensing in SRTF1-P22 Combined Cassette.png

In this graph, SRTF1 cassette (BBa_K3889100) and P22 Cassette(BBa_K3889101) were co-expressed in Bacillus subtilis 168 consisting of SRTF1 with mCherry reporter and P22 Cassette with Azurite reporter under various concentrations of progesterone.

Unexpectedly, the mCherry expression decreased at higher concentrations of progesterone. Further experimental evidence is needed to determine the cause. Additionally, the Azurite expression increased with progesterone levels signifying that p22 protein production increased. Hence, there is evidence to suggest that SRTF1 and SRTF1 DNA binding sites were interacting as expected and progesterone sensing is taking place in Bacillus subtilis.

Results Table:

Progesterone Conc. (in nM) Azurite mCherry
0 11374 297821
16.5 17101 281770
33 17275 287161
82.5 29714 249459
165 45536 153234
330 78069 117241


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]


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