Description:
pTWIN-MBP1 has the E. coli maltose binding protein (2) fused between the modifiedSsp DnaB (3) and Mxe GyrA inteins (4). The presence of the chitin binding domain from Bacillus circulans (5,6) facilitates purification. The pTWIN-MBP1 vector permits the isolation of a circular MBP species (1). The double-stranded vector is 8,518 base pairs in length.
Source:
pTWIN-MBP1 contains two mini-inteins, one derived from the Synechocystis sp DnaB intein (154 amino acids) (7) and the other from the Mycobacterium xenopi GyrA intein (198 amino acids) (8).
Advantages:
• A pBR322 derivative.
• Expression of the fusion gene is under the control of the T7 promotor (9) and is regulated by IPTG due to the presence of a lacI gene.
• Expression requires an E. coli host that carries the T7 RNA Polymerase gene [e.g., T7 Express Competent E. coli (High Efficiency) (NEB #C2566), BL21(DE3) Competent E. coli (NEB #C2527) and derivatives].
• Origin of DNA replication from the bacteriophage M13 allows for the production of single-stranded DNA by helper phage superinfection of cells bearing the plasmid.
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