title: Construction and expression of hybrid plasmids containing the Escherichia coli glyA genes.
abstract: The Escherichia coli glyA gene, encoding serine transhydroxymethylase (STHM), has been cloned in the plasmid vector pACYC184. The recombinant plasmid (pGS1) contains a 13 kb EcoRI insert. Genetic and biochemical experiments indicate that the region controlling STHM synthesis is present on the insert. Strains bearing multi-copy plasmid vectors carrying the glyA gene overproduce the enzyme from 17- to 26-fold. The glyA gene was identified on the insert by analyzing a set of plasmids derived from pGS1 that carry random insertions of the transposable kanamycin resistance element Tn5. Cloning of segments of the original insert into the plasmid pBR322 established that a 2.5 kb SalI-BclI fragment carries the glyA gene. A physical map of this fragment is presented.
mesh_terms: D003001:Cloning, Molecular / Q000379:methods; D004926:Escherichia coli / Q000235:genetics*; D005786:Gene Expression Regulation; D005998:Glycine / Q000235:genetics*; D012696:Glycine Hydroxymethyltransferase / Q000235:genetics*; D009876:Operon; D010957:Plasmids; D014166:Transferases / Q000235:genetics*
chemical_list: D014166:Transferases; D012696:Glycine Hydroxymethyltransferase; D005998:Glycine