Part:BBa_K3771075

Ptrc-CDO1

Description

P_trc-cdo1 is a composite part consisting of the trc promoter and the cdo1 sequences. This part was used in in vivo testing of taurine production. The sequence for cdo1 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 CDO1 enzyme.

Fig. 1 Confirmation of cdo1 fragment by PCR. M: Marker; Lane 1: cdo1 (603 bp)

Fig. 2 Confirmation of pSB4KI-Ptrc-CDO1 by digestion. M: Marker; Lane 1~3: Different colonies of pSB4KI-Ptrc-CDO1 (5470 bp)

Fig. 3 Transformation / CDO1 in DH5α

Fig. 4 Confirmation of protein expression of CDO1. M: Marker; Lane1: CDO1 in DH5α without induction; Lane2: CDO1 in DH5α with induction (~22 kDa)

Usage

CDO1 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. 5 Taurine production of Ptrc-cdo1 +PlacI-csad +Ptrc-cs in different growth mediums.

Fig. 6 Taurine production of Ptrc-cdo1+Ptrc-csad in different growth mediums.

Fig. 7 Taurine production (extracellular) of CDO1 and CSAD in ∆tauD.

Fig. 8 Taurine production (extracellular) of CDO1 and CSAD in DkO.

Fig. 9 Taurine production (intracellular and extracellular) of CDO1 and CSAD in DkO and top10.

Fig. 10 Taurine production (extracellular) of CDO1 and CSAD with expression of CSAD regulated by soxS promoter in DkO.

Characterization

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 NgoMIV site found at 534
  
• 1000
  COMPATIBLE WITH RFC[1000]