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| pAB-6xHis™ | Cat. #B4 | |
| DNA Sequence | ||
| pAB-6xHis™ plasmid transfer vector is designed for expression of proteins with N-terminal polyhistidine tag. A PCR fragment encoding a protein of interest can be cloned in lieu of the polyhedrin promoter and polyhistidine tag using any of the unique restriction endonuclease sites (SmaI, XbaI, EcoRI, NotI, PstI, BglII) located in the MCS. Proteins are expressed under control of strong very late polyhedrin promoter in the polyhedrin site of baculovirus DNA. Affinity purification of the expressed proteins can be achieved using Ni-NTA (nickel-nitrilotriacetic acid) resin (QUIAGEN). If desired, polyhistidine tag can be cleaved with thrombin at thrombin cleavage site (LVPRGS) encoded at nucleotides 4174-4191 of the vector. Unique SpeI site can be used to obtain 6xHis-tagged proteins without the thrombin site. Unique BamHI site can be use to obtain proteins without polyhistidine tag. Positions of phF and mR primers, which are useful for sequencing the insert/vector junctions, are shown on the DNA sequence as arrows. | ||
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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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