Part:BBa_K3771077

Ptrc-CSAD

Description
P_trc-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.

Fig. 1. Taurine pathways in E. coli

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).

Characterization

Fig. 2. Confirmation of csad fragment by PCR. M: Marker; Lane 1: csad (1368 bp)

Fig. 3. Confirmation of pSUI-P_trc-csad by digestion. M: Marker; Lane 1~3: Different colonies of pSUI-P_trc-csad (3674 bp)

Fig. 4. Transformation/CSAD in DH5α

Fig. 5. Confirmation of protein expression of CSAD. M: Marker; Lane 1: whole cell of CSAD in DH5α; Lane 2: soluble protein of CSAD in DH5α (~50 kDa)

References
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
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 337
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 122