Invitrogen™ Bac-to-Bac™ HBM TOPO™ Secreted Expression System

Description

• Bac-to-Bac™ Baculovirus Expression is an efficient method for producing baculovirus for high-level protein expression in insect cells
• It relies on generation of recombinant virus by site-specific transposition in E
• coli (DH10Bac™) rather than homologous recombination in insect cells
• The new pFastBac™ HBM TOPO™ vector enables secreted protein expression because it has the honeybee melittin (HBM) secretion signal
• The new vector should be strongly considered for the expression of glycoproteins
• Glycoproteins cannot be glycosylated in the absence of any secretion signal
• In contrast to glycoproteins secreted from mammalian cells, glycoproteins secreted from baculovirus can be easily de-glycosylated in vitro
• This is a very important feature in order to crystallize proteins! The pFastBac™ HBM TOPO™ vector has also a C-terminal His-Tag with a TEV cleavage signal to enable easy purification of Histidine fusion proteins on nickel-chelating resins (ProBond™ Purification System) and native proteins with the aid of AcTev™ protease
• The vector uses the strong polyhedrin promoter to generate high levels of expression in a variety of insect cell lines such as Sf9, Mimic™ Sf9, Sf21, and High Five™ cells in Sf-900 II & Sf-900™ III media from Gibco
• Traditional baculovirus systems require purification and amplification of an initial low-titer viral supernatant
• This requires a time-consuming plaque assay
• Bac-to-Bac™'s pFastBac™ vectors, however, recombines with the parent bacmid in DH10Bac™ E
• coli competent cells to form an expression bacmid
• Transfect the bacmid into insect cells for fast production of a high titer of pure recombinant baculovirus particles in the very first transfection
• You'll save weeks of precious time
• Collect a pure P2 baculovirus stock on Day 10 without the necessity of tedious plaques assay.