Part:BBa_K3771080

T7-CoaBC

Description
P_T7-coaBC is a composite part consisting of the T7 promoter and the coaBC sequences. This part was used in in vivo testing of taurine production. The sequence for jju and coaBC promoter were ligated and transformed into E. coli to calculate taurine production using high-performance liquid chromatography (HPLC).

Biology
T7 promoter constitutively facilitates the expression of CoaBC enzyme.

Fig. 1. Taurine pathways in E. coli

Usage

CoaBC enzyme was used in vitro testing of taurine production. The sequence for CoaBC 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 coaBC fragment by PCR. M: Marker; Lane 1: coaBC (1221 bp)

Fig. 3. Transformation/CoaBC in DH5α

Fig. 4. Confirmation of pET28a-P_T7-coaBC by digestion. M: Marker; Lane 1~3: Different colonies of pET28a-P_T7-coaBC (5398 bp)

Fig. 5. Transform into BL21(DE3)

Fig. 6. Confirmation of protein expression of CoaBC. M: Marker; Lane1: whole cell of CoaBC in BL21(DE3); Lane2: soluble protein of CoaBC in BL21(DE3) (~45 kDa)

References
BRENDA - Information on EC 4.4.1.10 - cysteine lyase. Brenda-enzymes.org. Published 2021. Accessed October 6, 2021. https://www.brenda-enzymes.org/enzyme.php?ecno=4.4.1.10
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 632
  
• 1000
  COMPATIBLE WITH RFC[1000]