What is baculovirus expression system?
Introduction. The baculovirus expression system (BEVS) has become a vital gene expression “tool” in the production and characterization of numerous recombinant proteins. Polyhedrin is a protein produced by wild-type baculovirus to protect the virus from environmental pressures such as temperature and osmolarity.
What is a Bac system?
Description. The Bac-to-Bac baculovirus expression system enables the efficient production of recombinant baculovirus for expression testing in insect cells. The system relies on generation of recombinant baculovirus by site-specific transposition in E. coli rather than homologous recombination in insect cells.
What are Sf9 cells used for?
Sf9 cells, a clonal isolate of Spodoptera frugiperda Sf21 cells (IPLB-Sf21-AE), are commonly used in insect cell culture for recombinant protein production using baculovirus. They were originally established from ovarian tissue. They can be grown in the absence of serum, and can be cultured attached or in suspension.
How do you make Bacmid?
Bacmid DNA is washed 2 × with 500 μL ice-cold 70% ethanol and centrifugation at 17,000 × g for 10 min at 4°C. Discard supernatant and air-dry the bacmid pellets at room temperature (10–15 min) in a sterile laminar flow hood. Avoid extended drying as this can lead to difficulties in solubilizing the pellet.
What is a baculovirus vaccine?
The baculovirus–insect cell expression system is an approved system for the production of viral antigens with vaccine potential for humans and animals and has been used for production of subunit vaccines against parasitic diseases as well.
Can baculovirus infect human cells?
These results show that baculovirus can infect nondividing mammalian cells, which implies that the viral genome is able to penetrate the mammalian nuclear membrane.
What is a BAC cooling tower?
BAC participates in the ECC programme for cooling towers. Open cooling towers discharge heat from water-cooled systems into the atmosphere. The hot process water is distributed over a fill pack (heat transfer media) to interface with air blown by a fan through the cooling tower.
What is baculovirus and cellular DNA?
Baculoviruses are DNA viruses that infect insect cells. Use of baculovirus as a vector to introduce cDNAs into insect cells results in high expression of recombinant protein (up to 25 to 50% of total cellular protein, or 1 g of protein per 109 cells, although 10 to 100 mg of protein per 109 cells is more usual32 ).
What are CHO cells and what are they used for?
CHO cells are the most common mammalian cell line used for mass production of therapeutic proteins. They can produce recombinant protein on the scale of 3-10 grams per liter of culture.
How big is a Bacmid?
Notes: Recombinant bacmid is >135 kb in size. Since restriction analysis is difficult to perform with DNA of this size, PCR analysis is recommended to verify the presence of your gene of interest in the recombinant bacmid.
How do you make a dh10bac competent cell?
Start a 5ml culture SOB with Kanamycin 50ug/ml and Tetracycline 10ug/ml final concentration. Incubate at 37°C overnight. The day after, start a 100ml SOB culture with Kanamycin 50ug/ml and Tetracycline 10ug/ml final concentration and inoculate with 1ml from the 5ml culture.
Is recombinant flu vaccine safe?
Is this vaccine safe? Are there any side effects that could occur? In clinical studies, the safety of recombinant flu vaccines was comparable to that of other injectable flu vaccines.
Is the Baculovirus expression vector system a manufacturing platform?
The baculovirus expression vector system (BEVS) platform has become an established manufacturing platform for the production of viral vaccines and gene therapy vectors.
Which is better the expisf or the baculovirus system?
The ExpiSf Expression System achieves protein yields that are vastly superior to traditional insect platforms. In performance tests against other suppliers’ media, the ExpiSf system demonstrated greater protein expression, delivering 3x more protein. Figure 1. Recombinant protein titers in ExpiSf and other baculovirus expression systems.
How is the baculovirus modified for Transgene expression?
The baculovirus has been modified by engineering of a mammalian expression cassette for transgene expression in mammalian cells. Baculoviruses are non-replicating in mammalian cells and thus have an excellent safety profile combined with being well-tolerated by cells.
How is baculovirus used to express heterologous genes?
R ecombinant baculoviruses are widely used to express heterologous genes in cultured insect cells and insect larvae. For large-scale applications, the baculovirus expression vector system (BEVS) is particularly advantageous.
