title: Cloning and expression of the metE gene in Escherichia coli.
abstract: A lambda-transducing phage was isolated that contains the metE gene. This gene codes for N5-methyl-H4-folate:homocysteine methyltransferase (EC 2.1.1.14), an enzyme that catalyzes the terminal reaction in methionine biosynthesis. A 9.1-kb EcoR1 fragment of this phage, containing the metE gene, was then cloned into pBR325. This plasmid, pJ19, was used to transform Escherichia coli strain 2276, a metE mutant, and restore the MetE+ phenotype. Although the transformed cells produced large amounts of the metE protein in vivo, in vitro studies using pJ19 as template showed low synthesis of the metE protein.
authors.forename: J
authors.initials: J
authors.lastname: Chu
authors.forename: R
authors.initials: R
authors.lastname: Shoeman
authors.forename: J
authors.initials: J
authors.lastname: Hart
authors.forename: T
authors.initials: T
authors.lastname: Coleman
authors.forename: A
authors.initials: A
authors.lastname: Mazaitis
authors.forename: N
authors.initials: N
authors.lastname: Kelker
authors.forename: N
authors.initials: N
authors.lastname: Brot
authors.forename: H
authors.initials: H
authors.lastname: Weissbach
mesh_terms: D010582:Bacteriophage lambda / Q000235:genetics; D002499:Centrifugation, Density Gradient; D003001:Cloning, Molecular; D004262:DNA Restriction Enzymes / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics*; D005786:Gene Expression Regulation; D005798:Genes, Bacterial; D008780:Methyltransferases / Q000235:genetics; D008854:Microscopy, Electron; D010641:Phenotype; D010957:Plasmids; D014161:Transduction, Genetic
chemical_list: D008780:Methyltransferases; C022725:5-methyltetrahydropteroyltriglutamate-homocysteine S-methyltransferase; D004262:DNA Restriction Enzymes