| Novagen's baculovirus transfer plasmids are classified into two major categories: vectors in which the first ATG of the target gene(s) defines the start of translation of the open reading frame (ORF), and N-terminal fusion vectors that include an optimal translation initiation site followed by sequences encoding the His•Tag®, S•Tag™, or CBD•Tag™ peptides or AcNPV gp64 signal peptide for secretion. The vector-encoded tag peptides are useful for the universal protocols of detection and purification of target proteins in the absence of antibodies or another specific ligand. They may be removed from the expressed protein using proteases. All pBAC™ vectors provide the opportunity for in-frame C-terminal His•Tag fusions if desired. In addition, the secretion vector, pBACsurf-1, provides the opportunity for display of the target protein on the surface of virions when constructed in-frame with a copy of the downstream AcNPV gp64 glycoprotein (1). The configuration of restriction sites in the multiple cloning region of the pBAC transfer vectors allows direct subcloning of inserts from many pET bacterial vectors. Each of Novagen's pBAC plasmids (except pBACsurf-1) is available with a b-glucuronidase (gus) reporter gene for visual identification of recombinant plaques. These are designated as the pBACgus transfer plasmids. Because the gus gene is driven by the P6.9 promoter, recombinant viruses produce b-glucuronidase in infected cells from the so-called "late" phase of virus gene expression (12-18 hours), and their plaques turn blue when stained with X-Gluc. A BacVector® gus Recombinant Virus Stock is available as a control for both gus staining and for analyses of infected cell proteins (e.g., to compare to target gene expression). Several pBAC plasmids are available as prepared LIC vectors, ready for annealing with appropriately prepared inserts. The prepared vectors enable fusion of target genes at the most desirable position relative to the enterokinase or Factor Xa cleavage site following the vector-encoded fusion sequences. Inserts are placed such that vector-encoded sequences can be completely removed by protease cleavage. The pbac04x vectors are designed for co-expression of up to 4 genes in the same cell. These vectors are extremely useful for expression of multisubunit proteins, multiple copies of a gene, multiprotein complexes, and for studies of protein:protein interactions (2-4). The vector sequences are provided in GenBank format, with significant features indicated. To convert the sequence into other formats, first select the sequence and copy (Command-C on Mac or Ctrl-C on Windows). Next, click here to open the NIH ReadSeq web page in a new window. Paste the sequence into the input field (Command-V on Mac or Ctrl-V on PC), choose your favorite format from the dropdown menu, and hit the "submit" button. - Boublik, Y., Di Bonito, P., and Jones, I.M. (1995) Bio/Technology 13, 1079–1084.
- Weyer, U. and Possee, R.D. (1991) J. Gen. Virol. 72, 2967–2974.
- Belyaev, A.S. and Roy, P. (1993) Nucleic Acids Res. 21, 1219–1223.
- Belyaev, A.S., Hails, R.S., and Roy, P. (1995) Gene 156, 229–233.
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