Cloning and expression of the metE gene in Escherichia coli.
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.
Chu
J
J
Shoeman
R
R
Hart
J
J
Coleman
T
T
Mazaitis
A
A
Kelker
N
N
Brot
N
N
Weissbach
H
H
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
D008780:Methyltransferases; C022725:5-methyltetrahydropteroyltriglutamate-homocysteine S-methyltransferase; D004262:DNA Restriction Enzymes