Method for preparing vaccine vectors for swine from swine IgG1 Fc recombinant baculoviruses

A technology of recombinant baculovirus and vaccine vector, applied in the field of biomedicine, can solve the problems of unfavorable virus entering designated parts, danger, etc., and achieve the effects of improving immune efficacy, improving antagonistic ability, and improving transduction efficiency.

Active Publication Date: 2015-11-11
武汉中拓康明生物科技有限公司 +3
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  • Abstract
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AI Technical Summary

Problems solved by technology

However, baculovirus is a heterogeneous species to the body after all, and the body has an immune effect on it, which is not conducive to the virus entering the designated site. In order to solve this problem, a complement inhibitory molecule was int

Method used

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  • Method for preparing vaccine vectors for swine from swine IgG1 Fc recombinant baculoviruses
  • Method for preparing vaccine vectors for swine from swine IgG1 Fc recombinant baculoviruses
  • Method for preparing vaccine vectors for swine from swine IgG1 Fc recombinant baculoviruses

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Embodiment 1

[0036] 1. Construction and verification of baculovirus surface display vector

[0037] Using the AmMultiBac plasmid as a template, amplify GP64SP and GP64TM-CTD respectively, and add a 6×His tag and four restriction sites of NheI, SalI, SacI and EcoRI between GP64SP and GP64TM-CTD to facilitate detection and exogenous genes inserting GP64SP and GP64TM-CTD into the PFBDM transfer vector p10 promoter in sequence; using the pDsred-Monomer-C1 plasmid as a template, the amplified CMV-dsred fragment was inserted into the PFBDM transfer vector ph promoter; in order to verify the pSur-gp64 vector Whether the construction is successful, the enhanced green fluorescent protein EGFP is inserted into the multi-cloning site for inspection.

[0038] 2. Gene cloning

[0039] The VSVG-ED gene was amplified by PCR using the pMD2.G vector as a template and VSVGTM-F and VSVGTM-R as primers; the pUC57-pFc plasmid was synthesized by Jinweizhi Company; -F and E2-R were obtained by PCR amplificatio...

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Abstract

The invention discloses a method for preparing vaccine vectors for swine from swine IgG1 Fc recombinant baculoviruses, and belongs to the field of biological medicine. The method includes connecting cloned swine IgG Fc genes and swine fever virus E2 genes to baculovirus surface display vectors to obtain recombinant transfer vectors; enabling the recombinant transfer vectors to acquire recombinant baculovirus Bacmid vectors by means of Tn7 transposition; enabling the recombinant baculovirus Bacmid vectors to carry out transfection on insect cells by the aid of a liposome process, collecting culture supernatants to acquire recombinant baculoviruses, infecting the insect cells by the recombinant baculoviruses, and collecting cells to carry out Western blot analysis and confocal microscopic analysis on the cells; enabling recombinant baculovirus swine fever vectors to carry out vaccine efficacy tests on the swine. The method has the advantages that surface display swine IgG Fc recombinant baculoviruses prepared by the aid of the method is high in survival ability in swine serum and can be prevented from being removed by serum complements, and accordingly the contents of effective antigens of vaccine immunity can be increased.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to a method for preparing a porcine IgG1Fc recombinant baculovirus as a porcine vaccine carrier. Background technique [0002] Baculovirus (Baculovirus) is a virus with insects as its only host. It can be used as a biological insecticide or as an expression vector to express a large amount of foreign proteins in insect cells and prepare vaccines. Studies have found that the recombinant baculovirus carrying a mammalian promoter in mammalian cells can initiate the expression of downstream foreign genes, but the virus cannot proliferate in mammalian cells and has little toxicity to cells. Successfully transduced cells can be stably passaged and reproduced. Efficient expression of foreign genes, mammalian cells have better post-translational modification of proteins than insect cells, and the expressed protein structure is closer to natural proteins. Therefore, baculovirus can be...

Claims

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Application Information

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IPC IPC(8): C12N15/866C12N15/40C12N15/13A61K39/187
Inventor 杜恩岐黄光东刘阳坤张小莺张元元肖龙敬晓棋邢贞召赵毅
Owner 武汉中拓康明生物科技有限公司
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