protein-structure-cropped-img

Traditional baculovirus expression systems utilize tedious and time consuming site-specific transposition in E. coli or lengthy homologous recombination in insect cells to generate the recombinant baculovirus. In contrast, BaculoDirect™ Baculovirus Expression System uses a quick, 1 hour Gateway® recombination reaction to produce the necessary bacmid for transfection, saving days to produce recombinant baculovirus.

Advantages of the BaculoDirect™ System:

  • Fast method generates recombinant virus in minimal time
  • Strong polyhedrin promoter produces mgs of protein
  • C-terminal or N-terminal 6xHis and V5 tag
  • Flexible Gateway® cloning allows use of any Entry vector

Fastest method to generate recombinant baculovirus

BaculoDirect™ Baculovirus Expression System makes baculovirus expression more convenient, requiring less hands on time than traditional systems. Baculovirus expression systems typically require bacterial transformation and isolation of a large bacmid or co-transfection of a transfer vector and linear baculovirus DNA into insect cells. The BaculoDirect™ system accelerates research by eliminating these time-consuming steps, saving you precious hands-on time.

BaculoDirect™ method overview

560-wide.par.2705.image.360.386.1

Our engineered BaculoDirect™ linear DNA contains attR sites for recombination of your gene of interest cloned into a Gateway® Entry clone. Simply mix the Entry clone with the BaculoDirect™ Linear DNA and Gateway® LR Clonase™ enzyme, incubate for 1 hour, and then transfect either Sf9 of Sf21 insect cell to produce recombinant virus.

Engineered for expression

560-wide.par.88719.image.360.177.1

The BaculoDirect™ linear DNA is designed for simple generation of recombinant baculovirus and expression insect cells. In addition to attR site for quick Gateway® recombination cloning, the backbone contains strong polyhedrin promoter for high protein expression and a C-terminal or N-terminal 6xHis and V5 tag for detection and purification. Show here at right is western blot analysis of 5 proteins cloned into and expressed using the BaculoDirect™ system.

Negative selection ensures viral stock purity

image.par.21235.image.180.109.1

A one-hour Gateway® LR reaction was performed using 300 ng of BaculoDirect™ Linear DNA and 100 ng of a Gateway® entry clone containing GFP. Ten Microliters of the LR reaction was used to transfect 2 x 106 Sf21 cells with Cellfectin® Reagent. Cells were grown 72 hours in Grace's Medium supplemented with 10% FBS and 100 µM ganciclovir.

Ten microliters of the resulting supernatant was used to infect 2 x 106 Sf21 cells. Cells were again grown in Grace's Medium supplemented with 10% FBS and 100 µM ganciclovir. After 72 hours, cells were examined for expression by fluorescence. β-gal staining was performed on the cells and no background of non-recombinant virus was performed.