What is a polyhedrin promoter?
As has been already mentioned above, the polyhedrin promoter is by far the most frequently exploited promoter for recombinant protein expression using baculovirus vectors. Initially, transfer vector pAc373 (Smith et al., 1985) was used to target the AcNPV polyhedrin gene locus for foreign gene expression.
What is baculovirus vector system?
The Baculovirus Expression Vector System (BEVS) is used to produce recombinant proteins. A baculovirus is an enveloped double-stranded DNA virus that belongs to the Baculoviridae family, which affects approximately 600 different insect species.
How do you concentrate on baculovirus?
Baculovirus can be concentrated by ultracentrifugation or tangential flow filtration (TFF) with limited impact on its infectivity.
What is the pathogenic property of baculovirus?
“What is the pathogenic property of baculovirus, used as a biological agents? Name the genus of these organisms.” Baculoviruses are pathogenus that attack insects and other arthopods. The majority of baculoviruses used as biological control agents are in the genes “Nucleopolyhedrovirus”.
Why baculoviruses are excellent biological control agents?
Baculoviruses have a narrow spectrum of activity. Because of this they can be used to control the specific pest insect without having an effect on non-target organisms, including pollinators, in the surrounding environment.
What type of virus is the baculovirus expression system?
Traditional baculovirus expression systems are based on an in vivo (cultured insect cells) low frequency recombination event between a shuttle vector carrying the gene of interest and the circular Autographa californica nuclear polyhedrosis virus (AcMNPV) double-stranded (ds) DNA viral genome.
How will you develop baculovirus vector?
There are two commonly used strategies, by direct insertion of foreign genes into a baculovirus shuttle vector (bacmid) in Escherichia coli [4] or co-transfection of insect cells with linearized baculovirus DNA (or replication-deficient bacmid) and a transfer plasmid carrying the gene of interest [5,6,7], for the …
Does the AcNPV polyhedrin promoter promote hypertranscription?
Hypertranscription and temporal expression from the Autographa californica nuclear polyhedrosis (AcNPV) baculovirus polyhedrin promoter involves an alpha-amanitin-resistant RNA polymerase and requires a trans-acting viral factor(s). We previously reported that a 30-kDa host factor, polyhedrin promot …
Is SV40 polyA required for the proper gene expression under the polyhedrin promoter?
In the baculovirus prototype Autographa californicamultiple nucleopolyhedrovirus (AcMNPV), the polyhedrin promoter (very late promoter) transcribes its gene by a viral RNA polymerase therefore there is no supporting evidence that SV40 polyA is required for the proper gene expression under the polyhedrin promoter.
Are insect specific baculoviruses useful for high yield protein expression?
The insect specific baculoviruses in the family of Baculoviridaehave been widely used for high yield expression of heterologous proteins in insect cells for research and pharmaceutical applications [1,2,3,4].
What are the Rep and cap genes driven by baculovirus?
The rep and cap genes are driven by polyhedrin and p10 baculovirus promoter, which are dependent on the baculovirus RNA polymerase for their transcription [ 45] and thus should not be active in the mammalian cells following rAAV delivery. Still, one cannot preclude the possibility of recombination leading to functional rep or cap sequences.