Expression of normal and variant human hypoxanthine-guanine phosphoribosyltransferase in E. coli.
1. ECPR is a rapid and effective means for generating recombinant human HPRT. 2. The Bl21 (DE3) T7 polymerase/T7 promoter system provides high level expression of human HPRT constructs after induction of the T7 polymerase gene with IPTG. 3. Human HPRT constructs expressed in E. coli mimic the variant properties originally demonstrated in lymphoblast extracts from affected individuals. 4. Human HPRT expressed in E. coli can be rapidly purified to near homogeneity by a two step purification scheme.
Davidson
B L
BL
Department of Internal Medicine, University of Michigan Medical School, Ann Arbor 48109-0680.
Roessler
B J
BJ
Palella
T D
TD
D001483:Base Sequence; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D014644:Genetic Variation; D006801:Humans; D007041:Hypoxanthine Phosphoribosyltransferase / Q000172:deficiency / Q000235:genetics* / Q000302:isolation & purification; D008969:Molecular Sequence Data; D009154:Mutation; D016133:Polymerase Chain Reaction
D004247:DNA; D007041:Hypoxanthine Phosphoribosyltransferase