Difference between revisions of "Part:BBa K2239014"
(→‎Information contributed by NFLS_Nanjing (2022))
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1. Amplify the gdh gene by nested PCR with amplification system of 25μL, 10×buffer 2.5μL, dNTP 5nmol, primers 25 pmol each, ddH2O 17.3 μL, template DNA 2μL, Ex Taq 0.2U. The condition of the reaction is 94℃ for 5 min, 94℃ for 30s, 45℃ for 30s, 72℃ for 50s for 35 cycles, then 72℃ for 7 min.
 
1. Amplify the gdh gene by nested PCR with amplification system of 25μL, 10×buffer 2.5μL, dNTP 5nmol, primers 25 pmol each, ddH2O 17.3 μL, template DNA 2μL, Ex Taq 0.2U. The condition of the reaction is 94℃ for 5 min, 94℃ for 30s, 45℃ for 30s, 72℃ for 50s for 35 cycles, then 72℃ for 7 min.
 +
 
2. The primers used in the first reaction is GDHeF and GDHiR, and the primers used in the second reaction is GDHiF and GDHiR.
 
2. The primers used in the first reaction is GDHeF and GDHiR, and the primers used in the second reaction is GDHiF and GDHiR.
 +
 
3. The product of PCR is then tested by DNA agarose gel electrophoresis.
 
3. The product of PCR is then tested by DNA agarose gel electrophoresis.
  

Revision as of 14:54, 13 September 2022


GDH (glucose dehydrogenas)

GDH (glucose dehydrogenase) catalyzes the conversion of beta-D-glucose to D-glucono-1,5-lactone and back, as it converts NADP+ to NADPH and back. It is used to catalyze the oxidation of beta-D-glucose into D-glucono-1,5-lactone and reduce NADP+ into NADPH during the process.[1]


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
    COMPATIBLE WITH RFC[1000]


Information contributed by NFLS_Nanjing (2022)

Part information is collated here to help future users of the BioBrick registry.

Metadata:

  • Group: NFLS_Nanjing (2022)
  • Author: Yunshu Wang
  • Summary: Added information collated from existing scientific studies

Experimental approach:

1. Amplify the gdh gene by nested PCR with amplification system of 25μL, 10×buffer 2.5μL, dNTP 5nmol, primers 25 pmol each, ddH2O 17.3 μL, template DNA 2μL, Ex Taq 0.2U. The condition of the reaction is 94℃ for 5 min, 94℃ for 30s, 45℃ for 30s, 72℃ for 50s for 35 cycles, then 72℃ for 7 min.

2. The primers used in the first reaction is GDHeF and GDHiR, and the primers used in the second reaction is GDHiF and GDHiR.

3. The product of PCR is then tested by DNA agarose gel electrophoresis.

References

  • Ghiffary M R , Prabowo C , Sharma K , et al. High-Level Production of the Natural Blue Pigment Indigoidine from Metabolically Engineered Corynebacterium glutamicum for Sustainable Fabric Dyes[J]. ACS Sustainable Chemistry And Engineering, 2021.
  • Staiano M, Pennacchio A, Varriale A, Capo A, Majoli A, Capacchione C, D'Auria S. Enzymes as Sensors[J]. Methods in Enzymology, 2017