BaculoDirect™ N-Term Transfection Kit

Catalog number: 12562062

Invitrogen™  Related applications: Protein Expression

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Description

The BaculoDirect™ Baculovirus Expression System is a powerful and versatile eukaryotic system for high-level protein expression in insect cells. The combination of Gateway™ Technology with baculovirus expression makes the BaculoDirect System the fastest and easiest method for production of recombinant baculovirus.

How it works
BaculoDirect™ Linear DNA (the baculovirus genome) is engineered to include attR sites for quick and efficient recombination with a Gateway entry clone. The gene of interest is recombined from the entry clone into the BaculoDirect Linear DNA using a simple, one-hour LR reaction (Figure 1). The resulting reaction mix contains the recombinant baculovirus carrying the gene of interest and is used to transfect insect cells. The need for transforming bacteria and isolating a large bacmid or co-transfecting a transfer vector and linear baculovirus DNA into insect cells is eliminated. As a result, the hands-on time is greatly reduced. Purified baculovirus can be isolated in less than one week.

Gateway linear DNA
BaculoDirect Linear DNA is designed for rapid cloning with a Gateway entry clone and subsequent expression in Sf9 or Sf21 insect cells. The Linear DNA features:

• Strong polyhedrin promoter for high-level expression
• R-sites for efficient recombination with any attL-flanked Gateway entry vector
• TK gene for negative selection using ganciclovir
• C-terminal V5-His tag (BaculoDirect™ C-Term Expression Kit) or N-terminal V5-His tag (BaculoDirect™ N-Term Expression Kit) for detection with anti-V5 antibody and purification with nickel-chelating resin
• TEV protease cleavage site for removal of the V5-His tag following purification (BaculoDirect N-Term Expression Kit)
• LacZ gene, to ensure a pure baculovirus stock is generated, which is replaced by your gene of interest after LR recombinant reaction

Additional materials required, available separately: Gateway entry clone.

A selection guide for choosing the most appropriate Gateway entry vector for your application can be found at: www.thermofisher.com/Gateway.

For Research Use Only. Not for use in diagnostic procedures.

Specifications

Cleavage: TEV Protease Recognition Site
Promoter: Polyhedrin
Vector Type: BaculoDirect Vectors
Product Size: 5 transfections
Cloning Method: Gateway®
Protein Tag or Fusion: His Tag (6x), V5 Epitope Tag

Contents & storage

The BaculoDirect™ transfection kits include five 0.3 µg vials of BaculoDirect Linear DNA, 125 µL Cellfectin transfection reagent, and ganciclovir.

Store the BaculoDirect Linear DNA and Cellfectin at 4°C. All components are guaranteed stable for 6 months if stored properly.

Documents

Manuals & protocols

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