Document Detail


Optimal production and in vitro activity of recombinant endostatin from stably transformed Drosophila melanogaster S2 cells.
MedLine Citation:
PMID:  11249205     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Recombinant plasmids containing a complementary deoxyribonucleic acid coding mouse endostatin were transfected and stably expressed in Drosophila melanogaster Schneider 2 (S2) cells. Stably transformed polyclonal cell populations expressing recombinant endostatin were isolated after 4 wk of selection with hygromycin B. Recombinant endostatin expressed in the stably transformed S2 cells under the influence of the Drosophila BiP protein signal sequence was secreted into the medium. Recombinant endostatin was also purified to homogeneity using a simple one-step Ni2+ affinity fractionation method. Purified recombinant endostatin inhibited endothelial cell proliferation in a dose-dependent manner. The concentration at maximum inhibition for recombinant endostatin was approximately 1.8 microg/ml. The stably transformed S2 cells produced 18 mg recombinant endostatin/L 7 d after induction with 5 microM CdCl2. Sodium butyrate supplementation (2.5 mM) increased recombinant endostatin production by 17%. These findings demonstrate optimal production and in vitro activity of recombinant endostatin from stably transformed D. melanogaster S2 cells.
Authors:
J H Park; K H Chang; J M Lee; Y H Lee; I S Chung
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  In vitro cellular & developmental biology. Animal     Volume:  37     ISSN:  1071-2690     ISO Abbreviation:  In Vitro Cell. Dev. Biol. Anim.     Publication Date:  2001 Jan 
Date Detail:
Created Date:  2001-03-15     Completed Date:  2001-06-07     Revised Date:  2008-11-21    
Medline Journal Info:
Nlm Unique ID:  9418515     Medline TA:  In Vitro Cell Dev Biol Anim     Country:  United States    
Other Details:
Languages:  eng     Pagination:  5-9     Citation Subset:  IM    
Affiliation:
Department of Genetic Engineering, Kyung Hee University, Suwon, South Korea.
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MeSH Terms
Descriptor/Qualifier:
Angiogenesis Inhibitors / biosynthesis*,  genetics
Animals
Blotting, Western
Butyrates / pharmacology
Cadmium / pharmacology
Cells, Cultured
Collagen / biosynthesis*,  genetics
DNA, Complementary / isolation & purification
Drosophila melanogaster / genetics*,  metabolism
Electrophoresis, Polyacrylamide Gel
Endostatins
Metallothionein / genetics
Mice
Peptide Fragments / biosynthesis*,  genetics
Promoter Regions, Genetic
Recombinant Proteins / biosynthesis,  genetics
Chemical
Reg. No./Substance:
0/Angiogenesis Inhibitors; 0/Butyrates; 0/DNA, Complementary; 0/Endostatins; 0/Peptide Fragments; 0/Recombinant Proteins; 7440-43-9/Cadmium; 9007-34-5/Collagen; 9038-94-2/Metallothionein

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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