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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
1
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pubmed:dateCreated |
1995-2-2
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pubmed:abstractText |
Shiga-like toxins (SLTs), which are produced by certain strains of Escherichia coli, are composed of enzymatically active A and B subunit multimers responsible for the toxin's binding. We have previously purified large amounts of the SLT-I B subunit by using a hyperexpression vector in Vibrio cholerae under the control of the trc promoter. In this study we examined various expression vectors to maximize yields of the SLT-II B subunit. The SLT-II B subunit has been expressed by using both the T7 promoter and the tac promoter in E. coli. When expressed from a plasmid containing the structural gene for SLT-II B deleted of the leader sequence, SLT-II B was able to form multimers when cross-linked, although SLT-II B production from this plasmid was unreproducible. SLT-II B expressed in all three systems appeared to form unstable multimers, which did not readily bind to a monoclonal antibody which preferentially recognizes B subunit multimers. SLT-II B expression was not increased by moving any of the plasmids into V. cholerae. Polyclonal antibodies raised to SLT-II B in rabbits recognized B subunit in SLT-II holotoxin yet were poorly neutralizing. SLT-II B was also expressed as a fusion protein with maltose-binding protein and could be cleaved from maltose-binding protein with factor Xa. Although the expression vectors were able to make large amounts of SLT-II B, as determined by Western blotting (immunoblotting), the levels of purified SLT-II B subunit were low compared with those obtained previously for SLT-I B subunit, probably because of instability of the multimeric SLT-II B subunit.
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pubmed:grant |
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pubmed:commentsCorrections |
http://linkedlifedata.com/resource/pubmed/commentcorrection/7806370-1741063,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7806370-2155181,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7806370-2201641,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7806370-2268304,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7806370-2644022,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7806370-2685131,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7806370-2807552,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7806370-3100913,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7806370-3231098,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7806370-3276522,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7806370-3284526,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7806370-3322851,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7806370-3519828,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7806370-6387911,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7806370-6392471,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7806370-8096622,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7806370-8253955,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7806370-8299898,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7806370-8321118,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7806370-8432592
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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 |
http://linkedlifedata.com/resource/pubmed/chemical/ATP-Binding Cassette Transporters,
http://linkedlifedata.com/resource/pubmed/chemical/Antibodies, Bacterial,
http://linkedlifedata.com/resource/pubmed/chemical/Bacterial Toxins,
http://linkedlifedata.com/resource/pubmed/chemical/Carrier Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Escherichia coli Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Maltose-Binding Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Monosaccharide Transport Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Recombinant Fusion Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Shiga Toxin 2,
http://linkedlifedata.com/resource/pubmed/chemical/maltose transport system, E coli
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pubmed:status |
MEDLINE
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pubmed:month |
Jan
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pubmed:issn |
0019-9567
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:volume |
63
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
301-8
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pubmed:dateRevised |
2010-11-18
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pubmed:meshHeading |
pubmed-meshheading:7806370-ATP-Binding Cassette Transporters,
pubmed-meshheading:7806370-Antibodies, Bacterial,
pubmed-meshheading:7806370-Bacterial Toxins,
pubmed-meshheading:7806370-Base Sequence,
pubmed-meshheading:7806370-Biological Assay,
pubmed-meshheading:7806370-Carrier Proteins,
pubmed-meshheading:7806370-Cloning, Molecular,
pubmed-meshheading:7806370-Escherichia coli,
pubmed-meshheading:7806370-Escherichia coli Proteins,
pubmed-meshheading:7806370-Genes, Bacterial,
pubmed-meshheading:7806370-Genetic Vectors,
pubmed-meshheading:7806370-HeLa Cells,
pubmed-meshheading:7806370-Humans,
pubmed-meshheading:7806370-Maltose-Binding Proteins,
pubmed-meshheading:7806370-Molecular Sequence Data,
pubmed-meshheading:7806370-Monosaccharide Transport Proteins,
pubmed-meshheading:7806370-Protein Conformation,
pubmed-meshheading:7806370-Recombinant Fusion Proteins,
pubmed-meshheading:7806370-Shiga Toxin 2,
pubmed-meshheading:7806370-Vibrio cholerae
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pubmed:year |
1995
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pubmed:articleTitle |
Expression and purification of Shiga-like toxin II B subunits.
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pubmed:affiliation |
Division of Geographic Medicine and Infectious Diseases, New England Medical Center, Boston, Massachusetts 02111.
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pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, P.H.S.,
Research Support, Non-U.S. Gov't
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