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Recombinant baculovirus vector, virus like particle, preparation method and use

A technology for recombinant baculovirus and baculovirus vector, which can be applied in the fields of botanical equipment and methods, biochemical equipment and methods, antiviral agents, etc., and can solve the problems of reduced transfection efficiency, limited vector capacity, and large IRES structure. , to achieve broad application prospects, high cutting efficiency, and good immunogenicity.

Active Publication Date: 2014-04-23
SOUTH CHINA UNITED VACCINE INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The system has the following defects: (1) The IRES itself has a large structure (about 0.5kb), and its application is often limited by the capacity of the vector, which will increase the burden on the vector and reduce the transfection efficiency
However, this expression method makes the cleaved VP1 have a 2A sequence of 20 amino acids, which affects its structure and immunogenicity
At the same time, the extra sequences have hidden dangers in quality standards and safety for human vaccines

Method used

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  • Recombinant baculovirus vector, virus like particle, preparation method and use
  • Recombinant baculovirus vector, virus like particle, preparation method and use
  • Recombinant baculovirus vector, virus like particle, preparation method and use

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Embodiment 1

[0072] A recombinant vector for expressing viruses disclosed in the present invention, one end of the 2A sequence of the picornaviridae is connected to the 3CD gene of the picornaviridae, the other end is connected to the P1 gene of the picornaviridae, and then the 3CD gene is placed P on the pFastBac Dual plasmid polh Under the promoter, obtain the recombinant vector Bac-P h -3CD-2A-P1.

Embodiment 2

[0074] Another recombinant vector for expressing viruses disclosed in the present invention is to connect one end of the 2A sequence of the picornaviridae to the 3C gene of the picornaviridae, and the other end to the P1 gene of the picornaviridae, and then connect the picornaviridae The 3C gene of the nucleic acid virus family is placed in the Pp of the pFastBac Dual plasmid olh Under the promoter, obtain the recombinant vector Bac-P h -3C-2A-P1.

Embodiment 3

[0076] Another recombinant vector for expressing viruses disclosed in the present invention is to connect one end of the 2A sequence of the picornaviridae to the 3C gene of the picornaviridae, and the other end to the P1 gene of the picornaviridae, and then connect the picornaviridae The 3C gene of the nucleic acid virus family is placed in the P of the pFastBac Dual plasmid 10 Under the promoter, obtain the recombinant vector Bac-P 10 -3C-2A-P1.

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Abstract

The invention discloses a recombinant baculovirus vector, a virus like particle, a preparation method and use and aims at providing tandem expression and accurate cutting expression of a plurality of genes with a single promoter and successfully packaging the virus like particle. The recombinant baculovirus vector is simple to operate, high in expression quantity and high in cutting efficiency. The obtained virus like particle is good in immunogenicity and does not have an exogenous sequence. The technical essential is that picornaviridae 3C / 3CD gene and P1 / P1op gene are expressed under the single promoter, the 3C / 3CD gene and the P1 / P1op gene are connected by a 2A sequence to build a 3C / 3CD-2A-P1 / P1op expression frame, then the 3C / 3CD gene is placed under the promoter of baculovirus vector plasmid to obtain a recombinant shuttle vector, a host cell is transfected to obtain recombinant baculovirus, and the host cell is infected to obtain the virus like particle.

Description

technical field [0001] The invention discloses a recombinant baculovirus vector body, and also discloses a virus-like particle comprising the virus recombinant vector, a virus-like particle, a preparation method and an application thereof, specifically, a virus expressing a Picornaviridae virus with high efficiency Recombinant vectors and virus-like particles. Background technique [0002] The Picornaviridae family includes 17 genera, most of which are highly pathogenic. Among them, Enterovirus, representative species: poliovirus (poliovirus poliovirus), hand, foot and mouth virus type 71 (Enterovirus71, EV71); rhinovirus (Rhinovirus), representative species: human rhinovirus A (Human rhinovirus A); Hepatovirus (Hepatovirus), representative species: hepatitis A virus (Hepatitis A virus, HAV); paraintestinal cytopathic human orphan virus (Parechovirus), representative species: human paraintestinal Human parechovirus, human parechovirus, is capable of causing severe human di...

Claims

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Application Information

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IPC IPC(8): C12N15/866C12N7/04A61K39/125A61P31/14
Inventor 王弋彭涛许昱华马书智安鸿尹海滨
Owner SOUTH CHINA UNITED VACCINE INST
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