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rdf:type |
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lifeskim:mentions |
umls-concept:C0017262,
umls-concept:C0017982,
umls-concept:C0035647,
umls-concept:C0036025,
umls-concept:C0079460,
umls-concept:C0079866,
umls-concept:C0086418,
umls-concept:C0185117,
umls-concept:C0205145,
umls-concept:C0591833,
umls-concept:C2911684
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pubmed:issue |
6
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pubmed:dateCreated |
1986-9-16
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pubmed:abstractText |
Murine (m) and human (h) granulocyte--macrophage colony-stimulating factors (GM-CSF) have been expressed in large quantities in Saccharomyces cerevisiae using a secretion vector containing the promoter and leader sequences of the mating pheromone alpha-factor. Functionally active mGM-CSF was identified by a proliferation assay with a factor-dependent cell line and by a granulocyte--macrophage colony formation assay using bone marrow cells. The activity of hGM-CSF was confirmed by stimulation of granulocyte--macrophage colony formation using human cord blood cells. Murine GM-CSF with various apparent mol. wts (13, 18, 24, 34 and 40 kd, as well as a smear of higher mol. wts) was detected in yeast culture medium by protein blotting using a rat monoclonal antibody specific for the mGM-CSF N-terminal region peptide. Protein blotting using a rat monoclonal antibody specific for the hGM-CSF N-terminal region demonstrated that a 15.6-kd and higher mol. wt heterogeneous species were secreted. Mutations introduced at each of the two potential N-linked glycosylation sites in mGM-CSF showed that the 13-kd protein is not glycosylated and the major 18-kd protein is mainly glycosylated at the more C-terminal site, whereas the heterogeneous higher mol. wt species were not affected by the mutations. The N-terminal amino acid of the 13-kd protein was shown to be Ser which was four amino acids in the C-terminal direction from the fusion point.
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pubmed:commentsCorrections |
http://linkedlifedata.com/resource/pubmed/commentcorrection/3525148-2856931,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3525148-300377,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3525148-313822,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3525148-3484805,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3525148-3858863,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3525148-3871523,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3525148-3876930,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3525148-3902470,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3525148-3902570,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3525148-3923623,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3525148-5432063,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3525148-6190815,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3525148-6295879,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3525148-6336730,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3525148-6390681,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3525148-6420074,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3525148-6430565,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3525148-6606682,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3525148-6610831,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3525148-6757864,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3525148-6973347,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3525148-7263636
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pubmed:language |
eng
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pubmed:journal |
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pubmed:citationSubset |
IM
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pubmed:chemical |
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pubmed:status |
MEDLINE
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pubmed:month |
Jun
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pubmed:issn |
0261-4189
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:volume |
5
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
1193-7
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pubmed:dateRevised |
2009-11-18
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pubmed:meshHeading |
pubmed-meshheading:3525148-Amino Acids,
pubmed-meshheading:3525148-Animals,
pubmed-meshheading:3525148-Antibodies, Monoclonal,
pubmed-meshheading:3525148-Base Sequence,
pubmed-meshheading:3525148-Colony-Stimulating Factors,
pubmed-meshheading:3525148-Glycosides,
pubmed-meshheading:3525148-Humans,
pubmed-meshheading:3525148-Mice,
pubmed-meshheading:3525148-Molecular Weight,
pubmed-meshheading:3525148-Mutation,
pubmed-meshheading:3525148-Plasmids,
pubmed-meshheading:3525148-Saccharomyces cerevisiae
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pubmed:year |
1986
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pubmed:articleTitle |
Expression of murine and human granulocyte-macrophage colony-stimulating factors in S. cerevisiae: mutagenesis of the potential glycosylation sites.
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pubmed:publicationType |
Journal Article
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