Composite

Part:BBa_K2573000

Designed by: Amanda Kuang   Group: iGEM18_Waterloo   (2018-08-03)
Revision as of 20:30, 10 September 2018 by Froggyinthewater (Talk | contribs) (Usage and Biology)


MetE Coding Device

Usage and Biology

E. coli MetE codes for an enzyme that catalyzes the final step of de novo methionine biosynthesis without using an intermediate methyl carrier and in the absence of vitamin B12 (N5-methyl-H4-folate:homocysteine methyltransferasecatalyzes). Methionine is an essential amino acid required for E.coli growth.


A 2.4kb fragment containing the MetE gene from plasmid pSK397 was cloned into bba_J4450. This recombinant molecule was used to transform E.coli strain DH5 alpha, a metE mutant, to restore the MetE+ phenotype.


CHARACTERIZATION: By introducing the MetE gene into plasmid desired for insert it can be used to differentiate between E.coli cells that have successfully taken up the insert and those that have not in a medium lacking methionine.

REFERENCES: https://journals.plos.org/plosbiology/article?id=10.1371/journal.pbio.0030031 https://biocyc.org/gene?orgid=ECOLI&id=EG10584


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 1864
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
//cds/enzyme
//plasmid
Parameters
None