Part:BBa_K3771077
Ptrc-CSAD
Description
Ptrc-csad is a composite part consisting of the trc promoter and the csad sequences. This part was used in in vivo testing of taurine production. The sequence for csad and trc promoter were ligated and transformed into E. coli to calculate taurine production using high-performance liquid chromatography (HPLC).
Biology
trc promoter constitutively facilitates the expression of CSAD enzyme.
Usage
CSAD was used in in vivo testing of taurine production. The sequence for CDO1 enzyme and trc promoter were ligated and transformed into E. coli to calculate taurine production using high-performance liquid chromatography (HPLC).
Fig. 2 Taurine production of Ptrc-cdo1 +PlacI-csad +Ptrc-cs in different growth mediums.
Fig. 3 Taurine production of Ptrc-cdo1+Ptrc-csad in different growth mediums.
Fig. 4 Taurine production (extracellular) of CDO1 and CSAD in ∆tauD.
Fig. 5 Taurine production (extracellular) of CDO1 and CSAD in DkO.
Fig. 6 Taurine production (intracellular and extracellular) of CDO1 and CSAD in DkO and top10.
Fig. 7 Taurine production (extracellular) of CDO1 and CSAD with expression of CSAD regulated by soxS promoter in DkO.
Characterization
Fig. 1 Confirmation of csad fragment by PCR. M: Marker; Lane 1: csad (1368 bp)
Fig. 2 Confirmation of pSUI-Ptrc-CSAD by digestion. M: Marker; Lane 1~3: Different colonies of pSUI-Ptrc-CSAD (3674 bp)
Fig. 3 Transformation/CSAD in DH5α
Fig. 4 Confirmation of protein expression of CSAD. M: Marker; Lane1: whole cell of CSAD in DH5α; Lane2: soluble protein of CSAD in DH5α (~22 kDa)
References
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 337
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 122
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