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.
This part is the MetE gene cassette under the inducible LacI promoter. 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
- 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 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.
//plasmid
| None |