Part:BBa_K2573000

MetE Coding Device

Usage and Biology

Methionine is an essential amino acid for E.coli growth. Respectively, the E. coli MetE gene is essential to proper methionine synthesis. It encodes for an enzyme that catalyzes the final step of de novo methionine biosynthesis without using an intermediate methyl carrier. The absence of vitamin B12 is also required, as it functions as a repressor of MetE.

Biosynthesis of Methionine Mechanism

This part is the MetE gene cassette under the inducible LacI promoter; designed to be an inducible MetE coding device.

A 2.4kb fragment containing the MetE gene from plasmid pSK397 was cloned into Bba_J04450 via PCR and NEBuild Hi-Fi Assembly.

REFERENCES: Pejchal, Robert, and Martha L Ludwig. “Cobalamin-Independent Methionine Synthase (MetE): A Face-to-Face Double Barrel That Evolved by Gene Duplication.” PLOS ONE, Public Library of Science, journals.plos.org/plosbiology/article?id=10.1371%2Fjournal.pbio.0030031.

“Escherichia Coli K-12 Substr. MG1655 MetE.” MetaCyc Parathion Hydrolase, 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]

Characterization of MetE Coding Device

We transformed the BioBrick into JT2-based E. coli strain and grew it in methionine deficient media. Since methionine is essential for E.coli growth and JT2 does not contain MetE in its genome, seeing growth indicates that the part does indeed work as intended as the JT2 cells, lacking methionine, would not have grown otherwise.