pAB-MBP™ vector encodes MBP tag derived from the E.coli malE gene (NCBI Acc# J01648). MBP tag is followed by a thrombin site and multiple cloning sites for insertion of a gene of interest in-frame with the tag. MBP/insert junction could be sequenced using MBPF forward primer 5'-GGTGAAATCATGCCGAACATC-3', which is complementary to the MBP tag sequence in vicinity of its C-terminus. The insert/vector junction can also be sequenced using MR reverse primer
5'-CGTGTCGGGTTTAACATTACGGATT-3' (indicated by the long arrow).
|pAB-MBP™ vector can be combined with any of the linearized baculovirus DNA vectors, i.e. ProEasy™, ProGreen™, ProFold™-ER1, ProFold™-C2, ProFold™-PDI and ProFold™-0. Click on TECHNOLOGY for more information on baculovirus expression system. Go to BACULOVIRUS TUTORIAL for simple on-line instructions on baculovirus transfection, propagation of recombinant baculoviruses and recombinant baculovirus protein expression studies for technical details.|
|The MBP fusion proteins are expressed under control of a very strong polyhedrin promoter. Since the level of expression is high, MBP fusion proteins are easily detectable as prominent bands after separating crude cell lysates in SDS-PAGE and staining with Coomassie blue.|
As documented in more than 100 studies, fusions of target proteins with maltose binding protein (MBP) tag can increase yield and solubility of the protein product. The MBP tag could be used either on the N- or C- terminus of the fusion proteins (Dyson M.R. et al., BMC Biotechnol., 4:32, 2004). Success rate can be dramatically increased for some low yield proteins, such as integral membrane proteins (Korepanova A. et al., Protein Expr Purif., 53:24-30, 2007). The fusion proteins could be purified on amylase resin available from New England Biolabs. The MBP tag could be cleaved by a protease, such as thrombin (ibid).
The above plasmid transfer vector is compatible with linearized baculovirus DNA from several other suppliers, e.g. BacMagic™, BacMagic™-2, BacMagic™-3, BacVector™-1000, BacVector™-2000, BacVector™-3000 (Novagen); BaculoGold™, BaculoGold™ bright (BD Pharmingen); flashBACGOLD™, flashBACULTRA™, flashBAC™ (OET); BacPAK6™ (Clontech); Sapphire™ (Orbigen). They are not compatible with Bac-to-Bac® and BaculoDirect™ DNA from Invitrogen. BacMagic™, BacMagic™-2, BacMagic™-3, BacVector™-1000, BacVector™-2000, BacVector™-3000 are trademarks of Novagen; BaculoGold™, BaculoGold™ bright are trademarks of BD Pharmingen; flashBACGOLD™, flashBACULTRA™, flashBAC™ are trademarks of OET; BacPAK6™ is a trademark of Clontech; Sapphire™ is a trademark of Orbigen, Bac-to-Bac® and BaculoDirect™ are trademarks of Invitrogen.
If you have not worked with baculovirus expression system, also referred to as baculovirus expression vector system or BEVS, you can get a quick update at TECHNOLOGY. Go to BACULOVIRUS TUTORIAL for simple on-line instructions on baculovirus transfection, propagation of recombinant baculoviruses and recombinant baculovirus protein expression studies.
Click on PRODUCTS to view the entire list of recombinant baculovirus-related products.
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