货号：12562054

品牌：ThermoFisher /Invitrogen/ Gibco/ ABI

规格：5 REACTION

目录价：￥22848.00

市场价格：￥20563.20

会员价格：￥20563.20

金山科研平台，产品价格货期咨询微信：jinshanbio 描述 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 WorksThe 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 directly to transfect insect cells. The need for transforming bacteria and isolating a large bacmid or co-transfection of 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 DNAThe BaculoDirect™ Linear DNA is designed for rapid cloning with a Gateway® entry clone and subsequent expression in Sf9 or Sf21 insect cells (Figure 2). 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• N-terminal V5-His tag for detection with Invitrogens Anti-V5 Antibody and purification with nickel-chelating resin (BaculoDirect™ N-Term Linear DNA)• TEV protease cleavage site for removal of the V5-His tag following purification (BaculoDirect™ N-Term Linear DNA)• C-terminal V5-His tag for detection with Invitrogens Anti-V5 Antibody and purification with nickel-chelating resin (BaculoDirect™ C-Term Linear DNA)Expression ResultsUsing the BaculoDirect™ System, expression can be detected as early as the first baculovirus infection. To demonstrate, green fluorescent protein (GFP) was expressed using BaculoDirect™ C-term. GFP expression was detected 72 hours post-infection (Figure 3). To ensure a pure baculovirus stock is generated, the BaculoDirect™ Linear DNA contains the lacZ gene. Following recombination with a Gateway® entry clone, an expression clone, and a by-product containing lacZ are generated. β-gal staining of the infected cell population indicated that the resulting baculovirus stock is pure and free of non-recombinant parental virus. 规格

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

内容及储存 The BaculoDirect™ Expression Kits include five 0.3 µg vials of BaculoDirect™ Linear DNA, 125 µl of Cellfectin® transfection reagent, frozen Sf9 cells, Gateway® LR Clonase® enzyme mix, GIBCO™ Unsupplemented Grace's Insect Medium, and ganciclovir. The BaculoDirect™ Transfection Kits include five 0.3 µg vials of BaculoDirect™ Linear DNA, 125 µl of Cellfectin® transfection reagent, and ganciclovir. Store the BaculoDirect™ Linear DNA, Cellfectin®, and Grace's Medium at +4°C. Store LR Clonase® mix at -80°C. Store Sf9 cells in liquid nitrogen. All components are guaranteed stable for 6 months if stored properly.