Coding

Part:BBa_K3771003

Designed by: KAO, YI-CHING   Group: iGEM21_NCKU_Tainan   (2021-09-30)
Revision as of 19:18, 18 October 2021 by Marvinang02 (Talk | contribs)


CSAD


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


Biology
lacI promoter constitutively facilitates the expression of CSAD enzyme.


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).



Characterization
The CDO1 fragment was synthesized by IDT and amplified by PCR. Agarose gel electrophoresis result is shown in Fig. 2.

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

Fig. 2 Confirmation of pSAA fragment by digestion. M: Marker; Lane 1: pSAA (2531 bp)

Fig. 3 Confirmation of protein expression of CDO1.M: Marker; Lane1: CDO1 in DH5α without induction; Lane2: CDO1 in DH5α with induction (~22 kDa)

Fig. 4 Taurine production of Ptrc-cdo1 +PlacI-csad +Ptrc-cs in different growth mediums.




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
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 250
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 35


[edit]
Categories
Parameters
None