Cloning & Expression

SBP
 

New SBP Bacterial System Expression Vectors
for Simple Purification and Detection Using Streptavidin Products

The new SBP (Streptavidin Binding Peptide) tag provides for one-step purification and detection of recombinant protein from bacterial lysates due to nanomolar affinity for streptavidin. SBP offers high-affinity, high yield, mild elution conditions and simplicity in recombinant protein purification. In addition, the tag provides versatility in downstream detection and assays due to the widely available streptavidin-conjugated fluorescent and enzymatic systems.

 

  • 38 amino acid tag (MDEKTTGWRGGHVVEGLAGELEQLRARLEHHPQGQREP)
  • One-step purification and detection
  • For use with widely available streptavidin products (resins, enzyme conjugates, plates, etc.)
  • Mild, specific elution of native protein using biotin
  • Higher purity than Maltose Binding Protein or HIS-tag1
  • Expression vectors containing the tac or T7 promoters

 


Bacterial Expression-tac Promoter

 

  • Cytoplasmic expression of C-terminal fusion
  • Ampicillin resistance, T1T2 transcriptional terminator, pMB1(derivative of pBR322) origin of replication, and lacI gene for repression of the tac promoter
  • Expression levels using the tac promoter may be in excess of 10 mg/L of culture when using IPTG as a de-repressor Expression in any established Escherichia coli expression host

 
Product MCS Region
P3996, pTAC-SBP-1
C-terminal SBP
pTAC-SBP-1

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Bacterial Expression-T7 Promoter

 

  • Cytoplasmic expression
  • N- or C- terminal fusions
  • Ampicillin resistance, T1T2 transcriptional terminator, pMB1 (derivative of pBR322) origin of replication, and lacI gene for repression of the T7 promoter
  • Higher yields of recombinant protein than tac system
  • lac operator (lacO) sequences reduce "leaky" expression
  • Requires hosts with a source of the T7 polymerase such as (DE3) lysogenic strains
  • Enterokinase cleavage of N-terminal SBP

 
Product MCS Region
P3746, pT7-SBP-1
C-terminal SBP
pT7-SBP-1
E9655, pT7-SBP-2
N-terminal SBP
pT7-SBP-2

Reference

 1.  Keefe, A. D., et al., Protein Expr. Purif., 23, 440-446 (2001).

 

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