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:
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.
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.