Abstract
Human prolactin was expressed in insect culture cells by recombinant baculoviruses carrying prolactin gene cDNA placed under the transcriptional control of polyhedrin gene promoter of Autographa californica nuclear polyhedrosis virus. Preliminary results of recombinant human prolactin expression as extracellular as well as intracellular product of baculovirus expression system were presented at the FEBS Meeting in Nice, France, in 1999 (Abstracts, p. 288). In the present work prolactin was expressed as a hexahistidine-tagged fusion protein and recombinant protein was purified by metal affinity resin. Yields varied between approximately 20 and 35 mg/liter of medium. This recombinant prolactin was biologically active in Nb2 lymphoma cell proliferation assay and after simple purification could substitute for pituitary-derived prolactin.
Copyright 2001 Academic Press.
MeSH terms
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Animals
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Baculoviridae / genetics
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Biological Assay
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Cell-Free System / chemistry
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Cell-Free System / metabolism
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Cells, Cultured
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Cloning, Molecular
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Growth Substances / biosynthesis
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Growth Substances / genetics
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Growth Substances / isolation & purification
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Growth Substances / physiology
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Humans
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Inclusion Bodies / genetics
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Inclusion Bodies / metabolism
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Intracellular Fluid / metabolism*
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Molecular Weight
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Nucleopolyhedroviruses / genetics
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Prolactin / biosynthesis*
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Prolactin / genetics*
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Prolactin / isolation & purification
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Prolactin / physiology
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Protein Denaturation
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Protein Renaturation
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Rats
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Recombinant Proteins / biosynthesis
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Recombinant Proteins / isolation & purification
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Recombinant Proteins / metabolism
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Recombinant Proteins / pharmacology
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Solubility
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Spodoptera / cytology
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Spodoptera / genetics*
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Spodoptera / metabolism*
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Spodoptera / virology
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Tumor Cells, Cultured
Substances
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Growth Substances
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Recombinant Proteins
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Prolactin