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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
5
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pubmed:dateCreated |
1995-3-9
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pubmed:abstractText |
This report describes a method whereby library mutagenesis combined with drug selection was used to generate unique and efficient ribosome-binding sites (RBS) for expressing recombinant proteins in Escherichia coli. The RBS was deleted from a vector expressing beta-lactamase and replaced with a 16-base sequence containing a library of mutations. Selection of the library with ampicillin yielded several unique RBS sequences that were more efficient than ompA RBS for expressing a bacterial (beta-lactamase) and a mammalian protein (single-chain Fv antibody). The described approach provides a practical means to improve recombinant protein expression and, also, provides new sequences to further evaluate the complex regulatory mechanism underlying translation initiation.
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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 |
Nov
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pubmed:issn |
0736-6205
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:volume |
17
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
944-53
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pubmed:dateRevised |
2003-11-14
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pubmed:meshHeading |
pubmed-meshheading:7840977-Animals,
pubmed-meshheading:7840977-Base Sequence,
pubmed-meshheading:7840977-Binding Sites,
pubmed-meshheading:7840977-Cloning, Molecular,
pubmed-meshheading:7840977-Electroporation,
pubmed-meshheading:7840977-Escherichia coli,
pubmed-meshheading:7840977-Gene Expression,
pubmed-meshheading:7840977-Gene Library,
pubmed-meshheading:7840977-Gene Transfer Techniques,
pubmed-meshheading:7840977-Immunoglobulin Fragments,
pubmed-meshheading:7840977-Mice,
pubmed-meshheading:7840977-Mice, Inbred BALB C,
pubmed-meshheading:7840977-Molecular Sequence Data,
pubmed-meshheading:7840977-Mutagenesis,
pubmed-meshheading:7840977-Mutagenesis, Insertional,
pubmed-meshheading:7840977-Polymerase Chain Reaction,
pubmed-meshheading:7840977-RNA, Ribosomal, 16S,
pubmed-meshheading:7840977-Recombinant Proteins,
pubmed-meshheading:7840977-Ribosomes,
pubmed-meshheading:7840977-beta-Lactamases
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pubmed:year |
1994
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pubmed:articleTitle |
Increased protein expression through improved ribosome-binding sites obtained by library mutagenesis.
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pubmed:affiliation |
Hybritech Incorporated, San Diego, CA.
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pubmed:publicationType |
Journal Article
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