title: Production of human plasma retinol-binding protein in Escherichia coli.
abstract: We designed a polymerase chain reaction (PCR) primer pair which allowed us to clone the cDNA coding for the human plasma retinol-binding protein (hRBP) into an Escherichia coli expression vector. Production of hRBP was confirmed by probing Western blots with antisera against plasma hRBP. Purification and characterization of the E. coli-produced plasma hRBP are also described. The availability of this expression system makes it possible to obtain large quantities of hRBP to facilitate our continuing studies of retinol and RBP metabolism.
authors.affiliation: Laboratory of Nutritional and Molecular Regulation, National Cancer Institute, Frederick Cancer Research and Development Center, NIH, MD 21702-1201.
authors.forename: T T
authors.initials: TT
authors.lastname: Wang
authors.forename: K C
authors.initials: KC
authors.lastname: Lewis
authors.forename: J M
authors.initials: JM
authors.lastname: Phang
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D015153:Blotting, Western; D003001:Cloning, Molecular; D017931:DNA Primers; D004926:Escherichia coli; D006801:Humans; D015151:Immunoblotting; D008969:Molecular Sequence Data; D010957:Plasmids; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D012177:Retinol-Binding Proteins / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D054839:Retinol-Binding Proteins, Plasma
chemical_list: D017931:DNA Primers; D011994:Recombinant Proteins; D012177:Retinol-Binding Proteins; D054839:Retinol-Binding Proteins, Plasma