pAcRP23-lacZ plasmid vector杆状病毒表达质粒载体 BioVector NTCC质粒载体菌种细胞基因保藏中心

pAcRP23-lacZ plasmid vector杆状病毒表达质粒载体

The flashBAC™ System is a streamlined platform for the production of recombinant baculoviruses.
Deletion of the essential gene ORF1629 from the Autographa californica nucleopolyhedrovirus
(AcMNPV) genome prevents non-recombinant virus from replicating within insect cells, thus
eliminating the need to plaque-purify recombinant virus from parental virus. Secreted or membranetargeted recombinant protein yields are greatly increased by the deletion of the chitinase gene (chiA).
Further nonessential gene deletions in flashBAC™ ULTRA provide enhanced quality and yield for
difficult to express proteins. The insertion of a bacterial artificial chromosome (BAC) at the
AcMNPV polyhedrin gene locus allows viral DNA to be maintained and propagated as a circular
genome within bacterial cells. The genomic DNA can then be purified and is the flashBAC™ DNA
provided with this kit.
To generate recombinant baculovirus using the flashBAC™ System, insect cells are transfected with
TransIT®-Insect Transfection Reagent, flashBAC™ DNA, and a transfer plasmid (e.g. pOET1,
pOET1C_6xHIS, or pOET6) containing the gene of interest. Homologous recombination within the
insect cells (see schematic at right) inserts the gene of interest under the control of the polyhedrin
promoter, and restores the function of ORF1629 allowing viral DNA to replicate and produce virus
particles. The recombinant virus genome replicates to produce baculovirus that can be harvested
directly from the culture medium of transfected insect cells. Because the flashBAC™ System has
effectively reduced recombinant baculovirus production to a one-step procedure, it is fully amenable to
high throughput and automated production platforms.

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