| Description: |
This plasmid expression
vector enables a protein of interest to be produced with a biotin acceptor
peptide (BAP) fused directly to its N-terminus, or, alternatively, as a fusion
to the C-terminus of E. coli maltose-binding protein with the BAP on its
N-terminus. |
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|
| Origin: |
A derivative of pMal-C2 (New
England Biolabs) |
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| Replicon: |
ColE1
(pBR322) |
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| Antibotic
Resistance: |
Ampicillin |
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| Promotor: |
trc |
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|
| Inducer: |
IPTG (1 mM) |
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| Notes: |
pDW363 also overproduces
biotin holoenzyme synthetase (BirA) to ensure efficient biotinylation of
BAP-tagged proteins in vivo. Add biotin to the medium (50 mM) when
cultivating cells that contain pDW363 or it derivatives. |
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|
| Literature
Citations: |
Tsao, K.-L., DeBarbieri, B.,
Michel, H. and Waugh, D. S. (1996). A versatile plasmid expression vector for
the production of biotinylated proteins by site-specific, enzymatic
modification in Escherichia coli. Gene 169: 59-64; Schatz. P. J. (1993).
Use of peptide libraries to map the substrate specificity of a
peptide-modifying enzyme: a 13 residue consensus peptide specifies
biotinylation in Escherichia coli. Bio/Technology
11:1138-43. |
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| Nucleotide
Sequence: |
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