A baculovirus expression vector

An expression vector and baculovirus technology, which is applied in the direction of virus/bacteriophage, virus, double-stranded DNA virus, etc., can solve the problem of unsatisfactory output of baculovirus expression vector system, high production cost of baculovirus expression system, etc. problems, to achieve high-yield characteristics, excellent production traits, and improved protein expression levels

Active Publication Date: 2022-07-15
NORTHWEST A & F UNIV
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Problems solved by technology

[0005] Expression system, the output of the baculovirus expression vector system is not satisfactory, which leads to high production costs of the baculovirus expression system

Method used

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  • A baculovirus expression vector
  • A baculovirus expression vector
  • A baculovirus expression vector

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Experimental program
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Effect test

Embodiment 1

[0024] 1. Knockout of Ac84-87 gene

[0025] Using primers (SEQ ID NO: 1 and SEQ ID NO: 2) with the upstream and downstream 50bp homology arms of the Ac84-87 DNA fragment, and using the pTriEx1.1 plasmid as a template, amplify the ampicillin resistance gene fragment to obtain 1023bp The PCR product (SEQ ID NO:3).

[0026] The obtained PCR product was transferred into Escherichia coli strain HS996 with RedET plasmid and Bacmid by electrotransformation, and was induced to produce recombinase by arabinose. The recombined Escherichia coli was screened by ampicillin resistance plate to obtain positive clones. At this point the Ac84-87 DNA fragment has been replaced by the ampicillin resistance gene fragment. Bacmid was extracted from Escherichia coli, identified by sequencing and named as BacmidΔAc84-87 (see appendix for the knockout strategy). figure 1 ).

[0027] BacmidΔAc84-87 was used for subsequent experiments after plasmid extraction and Bsu36I restriction endonuclease dige...

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Abstract

The invention discloses a novel baculovirus expression vector, which relates to the field of biotechnology. In the present invention, the novel baculovirus expression vector is obtained by simultaneously knocking out four consecutive non-essential genes Ac84, Ac85, Ac86 and Ac87 on the baculovirus expression vector. The novel baculovirus expression vector of the present invention increases the expression level of exogenous protein by 80%. At the same time, the proliferative properties of the virus are not affected. The increase in expression yield can significantly reduce the production cost of the enterprise. The baculovirus expression vector can be used in the biological product industry, especially in the subunit vaccine industry.

Description

technical field [0001] The present invention relates to the field of biotechnology, in particular to a baculovirus expression vector. Background technique [0002] Baculovirus is a double-stranded DNA virus that specifically infects arthropods, Spodoptera californica polynuclear polyhedrosis virus [0003] (Autographa californica nucleopolyhedrovirus, AcMNPV) is a type species of baculovirus. Since Smith GE et al. first used baculovirus to express the human β-interferon gene in insect cells in 1983 (MolCell Biol. 1983; 3:2156-65.), due to its low cost, high yield, and the availability of various translations Due to the advantages of post-modification system, baculovirus expression vector system is widely used in scientific research and production, especially in the fields of biological products and vaccine industries. SUMMARY OF THE INVENTION [0004] However, compared to the prokaryotic expression systems commonly used in industry (E. coli, Bacillus subtilis) and yeast ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/866C07K14/435
CPCC12N15/86C07K14/43595C12N2710/14143C12N2710/14134
Inventor 陈红英张潇月赵凯霞
Owner NORTHWEST A & F UNIV
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