Part:BBa_K3771078

Ptrc-CS

Description

P_trc-cs is a composite part consisting of the trc promoter and the cs sequences. This part was used in in vivo testing of taurine production. The sequence for cs 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 CS enzyme.

Fig. 1. L-cysteine sulfonic acid synthase catalyzes the reaction that turns O-phospho-L-serine into L-cysteine sulfonic acid. CDO1, L-cysteine dioxygenase; CSAD, L-cysteine sulfinic acid decarboxylase.

Usage

CS was used in in vivo testing of taurine production. The sequence for CS 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 P_trc-cdo1+P_lacI-csad +P_trc-cs in different growth mediums.

Characterization

Fig. 3. Confirmation of cs fragment by PCR. M: Marker; Lane 1: cs (1272 bp)

Fig. 4. Confirmation of pSUI fragment by digestion. M: Marker; Lane 1: pSUI (3664 bp)

Fig. 5. Transformation/CS in DH5α

Fig. 6. Confirmation of cs fragment by colony PCR. M: Marker; Lane 1~6: Different colonies of pSUI-P_trc-cs (1272 bp)

Fig. 7. Confirmation of pSUI-P_trc-CS by digestion. M: Marker; Lane 1: Colony of pSUI-P_trc-cs

Fig. 8. Confirmation of the protein expression of CS M: Marker; Lane 1: CS in DH5α with induction (~46 kDa)

References

Joo Y-C, Ko YJ, You SK, et al. Creating a New Pathway in Corynebacterium glutamicum for the Production of Taurine as a Food Additive. Journal of Agricultural and Food Chemistry. 2018;66(51):13454-13463. doi:10.1021/acs.jafc.8b05093
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 SapI.rc site found at 580