Coding

Part:BBa_K3331001

Designed by: Wang Weizhe   Group: iGEM20_XJTU-China   (2020-10-20)


E.coli galU

galU [Escherichia coli str. K-12 substr. MG1655] Encodes Glucose-1-phosphate uridylyltransferase, or UGPase, which catalyzes the production of UDP-glucose from glucose-1-phosphate and UTP. .This sequence comes from the genome of E.coli and we optimized the codon of this DNA sequence so that it could be expressed more efficiently in Bacillus subtilis.


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]


Characterization

We cloned and confirmed the target fragment by PCR. The length of our fragment is 981bp. The following results showed the successful result.

Figure 1:PCR confirmation of the target fragment.
T--XJTU-China--Bgimprove02.jpeg
T--XJTU-China--Bgimprove01.jpeg

the mRNA level of B.s GalU is ~2.4 fold higher than that of E.coli, indicating the significant improvement of enzyme expression and activity. However, due to the lower expression of another key enzyme PGM for EPS production in BE strain, the final EPS yield is lower than that of EE strain but still comparable.

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