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What is Baculovirus Expression System and its Advantages

2021-10-25
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What is Baculovirus Expression System

Baculovirus belongs to the Baculoviridae family (Baculoviridae), which is divided into nuclear polyhedrosis virus and granulovirus. The virus particles are rod-shaped and are a type of DNA virus that specifically infects arthropods and has an envelope. Genomic DNA is compressed and packed in a rod-shaped capsid in a super-coiled form, and the size is between 90 and 180 kb.
Baculovirus genome can be replicated and transcribed in the insect cell nucleus. After DNA replication, it is assembled in the baculovirus nucleocapsid. Because of its greater flexibility, it can accommodate large fragments of foreign DNA insertion and become large fragments for expression. Ideal carrier for DNA. Among them, the baculovirus currently used as a foreign gene expression vector is mainly nuclear polyhedrosis virus (NPV).

Medicilon researchers establish a well-developed baculovirus-insect cell expression services platform. We provide the expression and purification services on preparation of recombinant baculovirus and recombinant protein. We have a good track record in producing kinase and recombinant protein complexes.

Service:

  1. Generation of recombinant Bacmid DNA

  2. Preparation of recombinant baculovirus

  3. Titration of baculovirus

  4. Protein expression verification and optimization

  5. Small scale expression and purification of recombinant protein in insect cell

  6. Scale up expression and purification of recombinant protein in insect cell

Advantages of Baculovirus Expression System

As a currently widely used in vitro gene expression system, the insect baculovirus expression vector system has the following advantages over the prokaryotic expression system represented by Escherichia coli and the eukaryotic expression system including yeast and mammals:
(1) High safety: The host of baculovirus is limited to invertebrates, and it is not infectious to humans, animals and other vertebrates, and has strong specificity. Compared with Adenovirus vector and Lentivector That said, the security is good.
(2) High expression efficiency: The baculovirus very late polyhedrin promoter and P10 protein promoter are used to express foreign proteins strongly, and the expression level can reach up to 50% of the total protein content of the infected cells. The P10 protein is a non-essential segment in the viral genome, and the insertion of foreign genes does not affect the replication and infection of the virus.
(3) The expressed protein has biological activity: Compared with the prokaryotic expression system, insect cells are eukaryotic cells. The way they modify and process the protein after expression is similar to that of mammalian cells, and can recognize and correctly carry out signal peptides. Removal, phosphorylation, glycosylation and other reactions, and make the exogenous protein fold correctly, disulfide bond matching and oligomer formation in the cell, making it close to the natural protein in structure and function.
(4) When the foreign gene is inserted into the polyhedrin gene locus, it will inevitably cause the deletion or inactivation of the latter. Therefore, the recombinant virus will not be able to produce polyhedron. This not only provides a selection marker for the recombinant virus, but also the recombinant virus It cannot exist in the environment like wild viruses, and it is safer.
(5) Large capacity: The baculovirus genome is about 130Kb, with multiple strong natural promoters. It is also easy to construct new artificial promoters, which can express multiple genes at the same time and can accommodate large fragments of foreign genes.

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