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Article: Purification and physicochemical properties of a recombinant bovine growth hormone produced by cultured murine fibroblasts

TitlePurification and physicochemical properties of a recombinant bovine growth hormone produced by cultured murine fibroblasts
Authors
Issue Date1986
PublisherThe Endocrine Society. The Journal's web site is located at http://endo.endojournals.org
Citation
Endocrinology, 1986, v. 119 n. 4, p. 1489-1496 How to Cite?
AbstractMouse 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, Pdλ-pbGH 4-13, was found to secrete approximately 75 μg 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 (NH 4) 2SO 4 fractionation, ion-exchange chromatography, and gel filtration on Sephacryl S-200. The rbGH was characterized by bioassay, RIA, radioceptor 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 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 maps were also found to be similar between the rbGH and pituitary bGH preparations. The amino terminus of the rbGH showed similar heterogeneity to that of the bGH of puituitary origin. We conclude that rbGH which was synthesized, processed, and secreted from transformed mouse fibroblasts possessed almost exactly the same physicochemical properties as pituitary bGH.
Persistent Identifierhttp://hdl.handle.net/10722/178453
ISSN
2015 Impact Factor: 4.159
2015 SCImago Journal Rankings: 2.363
ISI Accession Number ID

 

DC FieldValueLanguage
dc.contributor.authorLeung, FCen_US
dc.contributor.authorJones, Ben_US
dc.contributor.authorSteelman, SLen_US
dc.date.accessioned2012-12-19T09:47:46Z-
dc.date.available2012-12-19T09:47:46Z-
dc.date.issued1986en_US
dc.identifier.citationEndocrinology, 1986, v. 119 n. 4, p. 1489-1496en_US
dc.identifier.issn0013-7227en_US
dc.identifier.urihttp://hdl.handle.net/10722/178453-
dc.description.abstractMouse 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, Pdλ-pbGH 4-13, was found to secrete approximately 75 μg 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 (NH 4) 2SO 4 fractionation, ion-exchange chromatography, and gel filtration on Sephacryl S-200. The rbGH was characterized by bioassay, RIA, radioceptor 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 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 maps were also found to be similar between the rbGH and pituitary bGH preparations. The amino terminus of the rbGH showed similar heterogeneity to that of the bGH of puituitary origin. We conclude that rbGH which was synthesized, processed, and secreted from transformed mouse fibroblasts possessed almost exactly the same physicochemical properties as pituitary bGH.en_US
dc.languageengen_US
dc.publisherThe Endocrine Society. The Journal's web site is located at http://endo.endojournals.orgen_US
dc.relation.ispartofEndocrinologyen_US
dc.titlePurification and physicochemical properties of a recombinant bovine growth hormone produced by cultured murine fibroblastsen_US
dc.typeArticleen_US
dc.identifier.emailLeung, FC: fcleung@hkucc.hku.hken_US
dc.identifier.authorityLeung, FC=rp00731en_US
dc.description.naturelink_to_subscribed_fulltexten_US
dc.identifier.doi10.1210/endo-119-4-1489-
dc.identifier.pmid3757900-
dc.identifier.scopuseid_2-s2.0-0022930259en_US
dc.identifier.volume119en_US
dc.identifier.issue4en_US
dc.identifier.spage1489en_US
dc.identifier.epage1496en_US
dc.identifier.isiWOS:A1986E254200010-
dc.publisher.placeUnited Statesen_US
dc.identifier.scopusauthoridLeung, FC=7103078633en_US
dc.identifier.scopusauthoridJones, B=8877805900en_US
dc.identifier.scopusauthoridSteelman, SL=6701781251en_US

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