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VP60 protein recombinant baculovirus carrying type O foot and mouth disease virus B cell epitope

A technology for recombining baculovirus and foot-and-mouth disease virus, which is applied in antiviral agents, virus/bacteriophage, and medical preparations containing active ingredients, etc. It can solve the problems of strong virus virulence, high production cost of inactivated vaccines, and loose toxins, etc. problem, to achieve the effect of improving immune efficacy, easy operation and good ability

Active Publication Date: 2012-07-25
JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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Problems solved by technology

But there are also shortcomings: for effective immunity, the antigen content of the vaccine must be guaranteed, the level of biosafety protection is high, incomplete inactivation may lead to loose virus, and the virus virulence of the vaccine will become stronger, etc. In addition, the production of inactivated vaccines higher cost

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  • VP60 protein recombinant baculovirus carrying type O foot and mouth disease virus B cell epitope
  • VP60 protein recombinant baculovirus carrying type O foot and mouth disease virus B cell epitope
  • VP60 protein recombinant baculovirus carrying type O foot and mouth disease virus B cell epitope

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Embodiment Construction

[0044] The present invention amplifies the capsid protein VP60 gene of rabbit hemorrhagic disease virus (without start codon) by PCR, and clones the PCR amplification product into the donor plasmid pFastBac of the baculovirus expression system TM In HTA, the recombinant transfer vector pFastBac was obtained TM HTA-VP60, and then insert the foot-and-mouth disease virus B cell epitope (200~213aa) into pFastBac TM Appropriate position of HTA-VP60 to obtain recombinant transfer vector pFastBac TM HTA-NF, use this vector to transform cells containing the shuttle plasmid Bacmid E. coli DH10Bac, the recombinant baculovirus plasmid Bacmid-NF was obtained through screening, and transfected into Sf9 cells to obtain the recombinant baculovirus rAcV-Bac-NF. Sf9 cells were infected with recombinant baculovirus to express the chimeric gene, and then the chimeric protein expressed by the recombinant baculovirus was used as an antigen to prepare the rabbit hemorrhagic disease virus-like p...

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Abstract

The invention relates to VP60 protein recombinant baculovirus carrying type O foot and mouth disease virus B cell epitope and application thereof, belonging to the field of biotechnology. The recombinant baculovirus is obtained by the following steps of: performing PCR (polymerase chain reaction) amplification on the rabbit hemorrhagic disease virus capsid protein VP60 (without initiator codon) gene; cloning the PCR amplification product into a pMD19-T vector and connecting into an eukaryon transfer vector; inserting the foot and mouth disease virus B cell epitope (200-213aa) to construct a baculovirus vector; and performing transfecting the Sf9 cell. The invention provides rabbit hemorrhagic disease virus capsid protein recombinant baculovirus carrying type O foot and mouth disease virus B cell epitope and rabbit hemorrhagic disease virus-like particles carrying type O foot and mouth disease virus B cell epitope (chimeric protein), wherein the chimeric protein is used as antigen to immunize mice, a positive antibody against type O foot and mouth disease virus VP1 protein can be induced and generated, and therefore, the chimeric protein can be used as antigen preventing and controlling the type O foot and mouth disease vaccine.

Description

[0001] technical field [0002] The invention relates to a rabbit haemorrhagic disease virus capsid protein VP60 recombinant baculovirus carrying an O-type foot-and-mouth disease virus B cell epitope and an application thereof, belonging to the field of biotechnology. Background technique [0003] Virus-like particles (VLPs) are composed of multiple copies of one or several viral structural proteins, mostly rod-shaped or icosahedral, with a diameter of about 25-100 nm. Both remain the same as live viral particles. VLPs are multimerized by one or several structural proteins, so their surface antigens are displayed in an orderly and repetitive manner, which can rapidly and significantly induce a high level of immune response. In addition, VLPs are moderate in size and can be effectively recognized and captured by antigen-presenting cells such as dendritic cells, thereby inducing a more effective immune response. VLPs are easy to purify by density gradient centrifugation or ...

Claims

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Application Information

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IPC IPC(8): C12N7/01C07K19/00A61K39/135A61P31/14G01N33/569
Inventor 王芳盛蓉范志宇胡波魏后军薛家兵姜平
Owner JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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