Cytoplasmic proteins | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Overexpressed cytoplasmic proteins are often misfolded which results in the accumulation of biologically inactive aggregates (inclusion bodies). In unmodified cells these proteins do not aggregate since they are expressed at low yield (typically below 0.1% of the total cell protein). At such low concentrations molecular chaperones are in excess to the newly synthesized proteins which ensures their correct folding. However, if a protein is overexpressed, its concentration by far exceeds concentration of molecular chaperones in the cytoplasm, resulting in target protein misfolding and aggregation. ProFold™-C1 and ProFold™-C2 linearized baculovirus vector DNA can provide for overexpression of both a cytoplasmic protein of interest and major cytoplasmic molecular chaperones Hsp40 and Hsc70. This often improves the yield of correctly folded cytoplasmic target proteins as compared to the same target proteins expressed using conventional vectors (Bac-to-Bac™**, BaculoGold™***, and BacPAK6™****) which do not provide molecular chaperones. Click on TECHNOLOGY for the details. |
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Mix and match features provided in the vectors | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Use pVL1393 plasmid transfer vector if you wish to express target proteins without affinity tags. Use pAB-6xHis™ plasmid transfer vector if you wish to express target proteins with a polyhistidine tag. Use pAB-GST™ plasmid transfer vector if you wish to express target proteins with a GST tag. Use pAB-MBP™ plasmid transfer vector if you wish to express target proteins with a maltose binding protein (MBP) tag. Note that an MBP tag could improve protein solubility. Combine any of the above plasmid transfer vectors with ProFold™-C1 linearized baculovirus DNA if you wish to provide Hsp40 and Hsc70 molecular chaperones. |
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The above features can be mixed and matched in various combinations, while supporting a high level of target protein expression. They can also be combined with "plain vectors" which only provide a high level of protein expression (pVL1393, ProEasy™, ProGreen™, ProFold™-0). A plasmid transfer vector is always combined with a baculovirus DNA vector, and some combinations are available as Kits. 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 details. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Examples | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
1.Solubility of a cytoplasmic protein is a problem. Insert gene encoding target cytoplasmic protein in pVL1393 plasmid vector if no affinity tag is required. Alternatively, use the pAB-GST™, pAB-6xHis™, or pAB-MBP™ vectors to provide target proteins with GST, His, or MBP tags. Co-transfect plasmid construct with ProFold™-C12 linearized baculovirus DNA. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
2.Target cytoplasmic protein is not problematic. Use ProEasy™ linearized baculovirus DNA, as it is the easiest method to construct recombinant baculoviruses. It is compatible with any of the pVL1393, pAB-GST™, pAB-6xHis™, or pAB-MBP™ plasmid transfer vectors. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
3. You have little experience with recombinant baculoviruses. Get a quick update on relevant recombinant baculovirus technologies at Materials and methods. Use ProEasy™ linearized baculovirus DNA, as it is the easiest method to construct recombinant baculoviruses. It is compatible with any of the pVL1393, pAB-GST™, pAB-6xHis™, or pAB-MBP™ plasmid transfer vectors. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Relevant products | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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*Go to PRODUCTS to order online. Alternatively, email an order as pdf attachment to info@abvector.com, or send it by Fax to 1-760-454-2465 or call 1-866-683-2867 to order by phone. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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. |