Tracer target plasmid for vaccinia virus Tian Tan TK gene and preparation method thereof

A vaccinia virus, gene technology, applied in the fields of botany equipment and methods, biochemical equipment and methods, genetic engineering, etc., can solve the problem of inability to analyze the distribution of the virus

Inactive Publication Date: 2018-01-19
XIAN MEDICAL UNIV
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  • Abstract
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  • Claims
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AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to provide a tracer targeting plasmid for the TK gene of the vaccinia virus Tiantan strain, which solves the problem that the existing recombinant vaccinia virus cannot analyze the distribution of the virus in the body

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  • Tracer target plasmid for vaccinia virus Tian Tan TK gene and preparation method thereof
  • Tracer target plasmid for vaccinia virus Tian Tan TK gene and preparation method thereof
  • Tracer target plasmid for vaccinia virus Tian Tan TK gene and preparation method thereof

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Embodiment Construction

[0038] The present invention will be described in detail below in conjunction with the accompanying drawings and specific embodiments. It should be understood that the specific embodiments described here are only used to explain the present invention, not to limit the present invention.

[0039] The experimental methods used in the following examples are conventional methods unless otherwise specified.

[0040] The materials and reagents used in the following examples can be obtained from commercial sources unless otherwise specified.

[0041] The genomic DNA sequence of the wild vaccinia virus Tiantan strain in the present invention is the sequence of GenBank No. AF095689.1 (VRL 14-FEB-2000).

[0042] The wild-type vaccinia virus Tiantan strain in the following examples is preserved in the Laboratory of Molecular Virology and Viral Immunology of Xi'an Medical College (this laboratory, the same below). Plasmid p-EGFP-Luc is preserved by our laboratory. Plasmid pMV was provi...

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Abstract

The invention discloses a tracer target plasmid for a vaccinia virus Tian Tan TK gene. The tracer target plasmid for the vaccinia virus Tian Tan TK gene comprises a starting vector, an upstream homologous recombination arm VTT-TKL of the vaccinia virus Tian Tan TK gene and a downstream homologous recombination arm VTT-TKR of the vaccinia virus Tian Tan TK gene; a vaccinia virus early/late promoterP11 and a late promoter P7.5 exist between the VTT-TKL and the VTT-TKR; a connecting sequence between the promoter P11 and the promoter P7.5 is provided with two restriction enzyme cutting sites; each of the downstream of the promoter P11 and the downstream of the promoter P7.5 comprises a poly-cloning site; a tracer gene Luciferase is inserted at the poly-cloning site behind the promoter P7.5; the nucleotide sequence of the tracer target plasmid is as shown in the sequence 1. According to the tracer target plasmid disclosed by the invention, the problem that the distribution situation of viruses in vivo cannot be analyzed by the existing recombinant vaccinia virus is solved. The invention also provides a preparation method for the tracer target plasmid.

Description

technical field [0001] The invention belongs to the technical field of biogenetic engineering, and in particular relates to a tracer targeting plasmid used for TK gene of vaccinia virus Tiantan strain, and also relates to a preparation method of the tracer target plasmid used for TK gene of vaccinia virus Tiantan strain. Background technique [0002] Vaccinia virus (vaccinia virus, VACV) belongs to Poxviridae, Orthopoxvirus genus, is an enveloped double-stranded DNA virus, and its genome is up to 200kbp. As a vaccine carrier or oncolytic virus, VACV has many unique advantages compared with other viral vectors such as retrovirus, human herpes simplex virus and adenovirus. Moreover, VACV, as a vaccine for human smallpox eradication, has been used in humans for more than a century, and its safety and effectiveness have been fully verified in practice. Therefore, VACV is considered to be an ideal oncolytic virus vector. [0003] Vaccinia virus Tian Tan (VACV-VTT) is a unique v...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/863
Inventor 马茜牛静燕周丽王婷汪洋
Owner XIAN MEDICAL UNIV
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