Part:BBa_K2573000
MetE Coding Device
Usage and Biology
Methionine is an essential amino acid for E.coli growth, and the MetE gene is essential for methionine synthesis. It encodes for an enzyme that catalyzes the final step of de novo methionine biosynthesis without using an intermediate methyl carrier. For optimal MetE function, vitamin B12 should not be present, as it functions as a MetE repressor.
This part is the MetE gene cassette under the inducible LacI promoter, designed to be a MetE coding device.
A 2.4kb fragment containing the MetE gene from plasmid pSKA397 was cloned into Bba_J04450 via PCR and NEBuilder HiFi DNA 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
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 1864
- 1000COMPATIBLE WITH RFC[1000]
Characterization of MetE Coding Device
We transformed the BioBrick into the JT2-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 the proper methionine synthesis pathway, would not have grown otherwise.
//plasmid
| None |