| Description: |
This derivative of E.
coli BL21(DE3) overproduces the catalytic domain of tobbaco vein mottling
virus (TVMV) protease in the form of an MBP fusion protein that cleaves itself
in vivo to yield a TVMV protease catalytic domain with an N-terminal
His-tag. |
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|
| Plasmid: |
The TVMV protease expression
vector pRK1035, 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: |
tac |
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|
| Inducer: |
IPTG (1 mM) |
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| Notes: |
The specificity of TVMV
protease is distinct from that of TEV protease. The canonical recognition site
for TVMV protease is ETVRFQS, with cleavage occurring between Q and S. Shifting
the temperature from 37 °C to 30 °C during induction maximizes the
yield of soluble TVMV protease. The TVMV protease open reading frame on pRK1035
was constructed from synthetic oligonucleotides and has been codon-optimized
for expression in E. coli. |
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|
| Literature
Citations: |
Yoon, H. Y., Hwang, D. C.,
Choi, K. Y., and Song, B. D. (2000). Proteolytic processing of oligopeptides
containing the target sequences by the recombinant tobacco vein mottling virus
NIa protease. Mol. Cells 10: 213-219. |
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| Nucleotide
Sequence: |
 |
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