BaculoDirect™ Baculovirus Expression System

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.

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.

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.

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.