Coding

Part:BBa_K4644003

Designed by: Han Cao   Group: iGEM23_BUCT   (2023-09-23)


ureEFGD

        Providencia rettgeri. UreEFGD protein coding region.This part enable the expression of the four accessory subunits of urease enzymes.

Usage and Biology

        ureEFGD are urease (accessory) proteins, responsible for the activation of the ureABC proenzyme.bacterial urease genes were widely reported in a way of urease gene cluster, which was consisted of structural genes and accessory genes. Structural subunits of urease were encoded by structural genes (ureABC), and accessory subunits of urease were encoded by accessory genes (ureEFGD).

        This part has three structural genes (ureA, ureB and ureC) and four accessory genes (ureE, ureF, ureG and ureD). It was also indicated that in addition to the above structural subunits of urease, accessory proteins are required in bacteria for the expression of active ureases. The genes encoding these proteins have been found in various arrangements adjacent to the structural genes. The protein products of the accessory genes (ureDEFG) are involved in the incorporation of nickel ions into the apoenzyme of urease.

        We constructed a related plasmid and expressed it.

Fig 1. PETlac-ureEFGD

Characterization

1. Agarose Gel Electrophoresis:

        As an important auxiliary structure of urease, we also obtained it from the genome of Providencia rettgeri strain JNB815 and recombined it into the pETlac plasmid using the same method..

Fig 2. The result of ureEFGD pcr.

Reference

Liu, X., Zhang, Q., Zhou, N. et al. Expression of an Acid Urease with Urethanase Activity in E. coli and Analysis of Urease Gene. Mol Biotechnol 59, 84–97 (2017).

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal SpeI site found at 975
    Illegal PstI site found at 1603
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1827
    Illegal SpeI site found at 975
    Illegal PstI site found at 1603
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 183
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal SpeI site found at 975
    Illegal PstI site found at 1603
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal SpeI site found at 975
    Illegal PstI site found at 1603
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
//chassis/prokaryote/ecoli
Parameters
biologyProvidencia rettgeri strain JNB815
proteinureEFGD