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Recombined sheep poxvirus transfer vector and construction method and application thereof

A sheeppox virus and transfer vector technology, applied in the field of genetic engineering, can solve problems such as increasing workload, and achieve the effects of good recombination efficiency, widening application scope and excellent screening efficiency

Inactive Publication Date: 2014-08-06
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI +1
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Problems solved by technology

This type of operation has certain limitations. When exogenous DNA needs to be replaced, the vector used for recombination must be rebuilt and identified, which increases a lot of workload.

Method used

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  • Recombined sheep poxvirus transfer vector and construction method and application thereof
  • Recombined sheep poxvirus transfer vector and construction method and application thereof
  • Recombined sheep poxvirus transfer vector and construction method and application thereof

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Embodiment Construction

[0056] The following examples facilitate a better understanding of the present invention, but do not limit the present invention. The experimental methods in the following examples are conventional methods unless otherwise specified. The test materials used in the following examples, unless otherwise specified, were purchased from conventional biochemical reagent stores.

[0057] 1 Materials and reagents

[0058] Escherichia coli DH5α strain, Trans2K plus II DNA Marker, purchased from Beijing Quanshijin Biotechnology Co., Ltd.; DS15000 DNA Marker, purchased from Guangzhou Dongsheng Biotechnology Co., Ltd.; restriction endonucleases HindⅢ, AvrⅡ, SalⅠ, SacⅠ, XhoⅠ, NheⅠ, NotⅠ, SacⅡ, SbfⅠ, SpeⅠ, AscⅠ, BamHⅠ, T4DNA ligase are NEB products; HS DNA Polymerase is a TaKaRa product; AxyPrep TM Plasmid Miniprep Kit, AxyPrep TM DNA Gel Extraction Kit is a product of AxyGEN; vector pVAX1 is a product of Invitrogen; pIRES2-AcGFP1 is a product of Clontech; lamb testis cells (Lamb test...

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Abstract

The invention provides a recombined sheep poxvirus transfer vector. A vector containing resistance genes and a plasmid replicon sequence and adopting Hind III and Avr II for digestion, such as a pVAX1 vector, is adopted, a DNA segment is inserted via introducing Hind III and Avr II enzyme cutting sites; the DNA segment comprises a gene sequence of a homologous arm with bidirectional poxvirus promoters and a sheep poxvirus TK gene. The invention further provides a recombined sheep poxvirus containing the recombined sheep poxvirus transfer vector and application of the recombined sheep poxvirus in the transfer of an exogenous antigen. Exogenous genes can be conveniently inserted into the recombined sheep poxvirus transfer vector, provided by the invention; moreover, the recombined sheep poxvirus transfer vector is improved according to a recombination result, and finally higher recombination efficiency is realized, accordingly, a good basis for the research and preparation of such recombination vaccines is laid.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and in particular relates to a recombinant sheep pox virus transfer vector and its construction method and application. Background technique [0002] Utilizing virus as a delivery system for exogenous antigens is a mature and effective vaccine design and development strategy, such as adenovirus live vectors, poxvirus live vectors, herpes virus live vectors, picornavirus live vectors, etc., have achieved good results as new vaccines. immune effect. Among them, due to its large and complex genome, poxvirus has the advantages of large capacity as a live vector and a large number of non-essential fragments for replication that can be used as sites for inserting foreign genes, so it has become an ideal vector for this type of vaccine research. Pox virus vectors that have been constructed so far include live vectors of vaccinia virus, fowl pox virus, swine pox virus, sheep pox virus and ra...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/86C12N15/66C12N7/01A61K39/12A61P31/12C12R1/93
Inventor 李继东才学鹏窦永喜朱学亮蒙学莲骆学农
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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