Baculovirus DNA Developments

 Our  research scientists have more than 30 years of experience on  baculovirus expression systems. Recently, Sheatech research group has  developed a highly quality version of baculovirus DNA, superBAC DNA. The superBAC DNA is developed and prepared in high quality DNA and confirmed with restriction enzyme digestion analysis, PCR and co-transfection etc.   


The superBAC DNA,  is derived from a wild type AcMNPV genome, contains an ORF 1629 partial  deletion which prevents non-recombinant virus from replicating in  insect cells. In addition, superBAC DNA  provides a positive selection for creating baculoviruses recombinants  in preparation for generating recombinant protein expression in insect  cells. The kit improves on the traditional method for generating  recombinant baculoviruses by eliminating the tedious, time-consuming  steps of plaque purification.
 

SuperBAC is  modified and integrated with folding protein genes, including PDI and  cdc37, which are regulated by the baculovirus promoter. The folding  protein genes remain in the recombinant virus genome after homologous  recombination in sf9 insect cells. Recently, superBAC derivatives, superBAC_PDI and superBAC_cdc37 with single modification, have also been constructed. 
 

 The superBAC BV DNA kit contains the superBAC DNA, Insect Transfection Reagent

and a positive control plasmid to monitor co-transfection efficiency.  

 

The advantages of superBAC DNA for co-transfection:


  • High quality DNA for co-transfection. 
  • No background: 100% recombinant virus. 
  • Direct amplification to increase virus titer for protein expression. 

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High Quality of Baculovirus DNA

 The following images were obtained from analysis experiments on  Sheatech’s new baculovirus genomic DNA. Baculovirus genomic DNA and  linear baculovirus genomic DNA were loaded onto a TAE agarose gel.  (L1=10 uL of baculovirus genomic DNA; L2= 5 ul of baculovirus genomic  DNA ; L3=2 ul of baculovirus genomic DNA ; L4=10 uL of linear  baculovirus genomic DNA; L5=5 uL of linear baculovirus genomic DNA ;  L6=2 uL of linear baculovirus genomic DNA; L7=DNA markers.) The result  shows high quality virus DNA, which was prepared by our scientists in  Sheatech.   

Baculovirus DNA RE mapping

 Restriction enzyme digested baculovirus DNA 

separated in 0.6% TAE agarose gel.   

Other Projects

DNA Assembly Cloning

T5 exo nuclease removed 5’end of double strand DNA

Single stand DNA annealed

Deep Vent DNA polymerase filled the gaps

Tag DNA ligase sealed the nicks at 50oC

Direct transform into DH5a competent cell

One step isothermal assembly reaction of linearized transfer vector and PCR fragments. From lane1 to lane4, each sample has one linearized transfer vector and one PCR product (PCR 1). From lane5 to lane8, each sample has one linearized transfer vector and two PCR products (PCR 1 & PCR 2). Both lane2 and lane6 are negative control samples. Lane4 sample is the negative control of lane8 sample. The DH5a transformed results were expected and colonies were confirmed by mini preparation and restriction enzyme analysis.