Table of Contents
What is polyhedrin promoter?
The polyhedrin gene promoter has an essential role in regulating foreign gene expression in baculovirus expression vector systems (BEVS); however, the high-level transcription mechanism is still unknown.
What is baculovirus vaccine?
Baculovirus is used as a vector vaccine candidate, in which the antigens can be expressed by the vector within the host cells and displayed on the baculovirus surface or plasma membrane.
What is the p10 promoter?
Fortunately, the p10 gene promoter can be duplicated in the virus genome upstream of the polyhedrin gene promoter to derive vectors (e.g. pOET5) for multiple gene expression. These expression cassettes can be duplicated so that several foreign genes can be inserted at the polyhedrin gene locus.
What is occlusion body?
Occlusion bodies are protein lattices that protect infectious particles after release into the environment. This strategy is mostly used by insect viruses to keep long-term infectivity of tissues after the host death.
What are occlusion bodies and where are they typically found?
What is baculovirus vector system?
The baculovirus vector system is widely used for the expression of recombinant proteins in cultured insect cells. It is one of the most versatile and powerful systems for eukaryotic expression of recombinant proteins.
How do you amplify baculovirus?
The Quick Amplification Guide suggests the best way to amplify, should the baculovirus yield be lower than average. Add 2ml PI stock to 50ml Sf9 cells at 2×106 cells/ml in a 1L Erlenmeyer flask. Add 2ml PI stock to 10ml Sf9 cells at 2×106 cells/ml in a 125ml Erlenmeyer flask.
How do you identify inclusion bodies?
As a first check, you could have a look at your culture with a microscope equipped with phase contrast illumination. When big enough, inclusion bodies appear as typical refringent granules (they could be mistaken for spores).
What are inclusion bodies made of?
Inclusion bodies are often made of denatured aggregates of inactive proteins. Although, the renaturation of inclusion bodies can sometimes lead to the solubilisation and the recovery of active proteins, the process is still very empirical, uncertain and of low efficiency.