title: Expression and partial purification of human prolactin in Escherichia coli.
abstract: 1. Human prolactin has been expressed in Escherichia coli. A cDNA fragment coding for the signal sequence and the full length prolactin molecule was cloned into the expression vector pUR291 which directs the synthesis of a beta-galactosidase prolactin fusion protein when expressed in E. coli. 2. Cultures of E. coli harbouring the recombinant plasmid pJMBG62 produced a fusion protein of the appropriate molecular weight which was detected by Western blot analysis using a polyclonal antibody raised against pituitary-derived human prolactin. 3. The fusion protein was isolated from inclusion bodies in a partially pure form and it was used as immunogen to raise antibodies against human prolactin. 4. When this partially purified fusion protein was injected into rabbits it generated antisera with good prolactin titres in animals which were rested for one year following a disappointing primary immunization with purified human prolactin.
authors.affiliation: School of Animal and Microbial Sciences, Department of Biochemistry & Physiology, University of Reading, Whiteknights, England.
authors.forename: M S
authors.initials: MS
authors.lastname: Gilbert
authors.forename: P J
authors.initials: PJ
authors.lastname: Lowry
authors.forename: M G
authors.initials: MG
authors.lastname: Castro
authors.forename: R J
authors.initials: RJ
authors.lastname: Woods
authors.forename: D
authors.initials: D
authors.lastname: Savva
mesh_terms: D015153:Blotting, Western; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics*; D015870:Gene Expression; D005796:Genes; D006801:Humans; D007106:Immune Sera; D010957:Plasmids; D011388:Prolactin / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D012045:Regulatory Sequences, Nucleic Acid; D015183:Restriction Mapping
chemical_list: D007106:Immune Sera; D011993:Recombinant Fusion Proteins; D011388:Prolactin