Purification and physiochemical properties of a recombinant bovine growth hormone produced by cultured murine fibroblasts

Endocrinology
F LeungJ J Kopchick

Abstract

Mouse fibroblast cell lines which secrete bovine (b) GH have been generated. This was accomplished by cotransforming mouse L cells (thymidine kinase-negative [TK-] and adenine phosphoribosyl transferase-negative [APRT-]) with DNA molecules encoding the Rous sarcoma virus-long-terminal repeat and bGH genes along with the herpes viral TK gene and the hamster APRT gene. One stable cell line, Pd lambda-pbGH 4-13, was found to secrete approximately 75 micrograms bGH per 24 h/5.0 X 10(6) cells. Media from this cell line were collected for purification of recombinant bGH (rbGH). Purification involved (NH4)2SO4 fractionation, ion-exchange chromatography, and gel filtration on Sephacryl S-200. The rbGH was characterized by bioassay, RIA, radioreceptor assay, and sodium dodecyl sulfate gel electrophoresis. Results of these analyses were compared with those obtained with a highly purified pituitary bGH. In the rat tibia bioassay, rbGH was found to be as potent as pituitary bGH. Results from the RIA, radioreceptor assay, and sodium dodecyl sulfate gel electrophoresis and Western blot analysis also suggested that the rbGH was similar to that of pituitary origin. Amino acid composition, partial (amino-terminal) sequence, and tryptic peptide ...Continue Reading

Citations

Sep 1, 1994·Comparative Biochemistry and Physiology. Part C, Pharmacology, Toxicology & Endocrinology·J A ProudmanL A Cogburn
Jun 1, 1997·Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology·D N FosterL K Foster
Aug 1, 1987·Proceedings of the National Academy of Sciences of the United States of America·C FinazD Dampfhoffer
Jul 1, 1990·Proceedings of the National Academy of Sciences of the United States of America·W Y ChenJ J Kopchick
Feb 15, 1994·Proceedings of the National Academy of Sciences of the United States of America·X WangJ J Kopchick
Sep 26, 2000·Molecular Genetics and Metabolism·J J Kopchick, J M Andry
Mar 21, 2007·Toxicological Sciences : an Official Journal of the Society of Toxicology·Georgia M FarrisSrinivasa Prahalada
Dec 31, 1990·Biochemical and Biophysical Research Communications·D N FosterL K Foster

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