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Tracking targeting plasmid of vaccinia virus Tiantan strain and preparation method thereof

A vaccinia virus and plasmid technology, applied in the field of biological genetic engineering, can solve the problems of untraceable recombinant VTT virus, lack of fluorescent screening markers, lack of tracer reporter genes, etc.

Pending Publication Date: 2019-08-23
XIAN MEDICAL UNIV
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Problems solved by technology

These targeting plasmids either lack a tracer reporter gene or a fluorescent selection marker, and the resulting recombinant VTT virus is either unable to be traced, or has a weak traceability, or is difficult to screen, so it cannot meet the requirements of the current attenuated oncolytic vaccinia virus. research needs

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  • Tracking targeting plasmid of vaccinia virus Tiantan strain and preparation method thereof
  • Tracking targeting plasmid of vaccinia virus Tiantan strain and preparation method thereof
  • Tracking targeting plasmid of vaccinia virus Tiantan strain and preparation method thereof

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Embodiment Construction

[0037] The present invention will be described in detail below in conjunction with the accompanying drawings and specific embodiments.

[0038] The materials and reagents used in the present invention can be obtained from commercial sources unless otherwise specified.

[0039] The genomic DNA sequence of the wild vaccinia virus Tiantan strain in the present invention is the sequence (VRL 14-FEB-2000) of GenBank number AF095689.1. The Tiantan strain of vaccinia virus was preserved in the Laboratory of Molecular Virology and Viral Immunology of Xi'an Medical College (this laboratory, the same below). Plasmids pEGFP-Luc and pMV-VTTTK-Luc are preserved by our laboratory.

[0040] One, a tracer targeting plasmid of vaccinia virus Tiantan strain, such as figure 1 As shown, including the pMV-VTTTK-ELuc plasmid, the pMV-VTTTK-Luc plasmid is inserted with the upstream homologous recombination arm VTT-TKL and the downstream homologous recombination arm VTT-TKR of the TK gene of the va...

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Abstract

The invention discloses a tracking targeting plasmid of a vaccinia virus Tiantan strain. The plasmid includes upstream homologous recombinant arms VTT-TKL and downstream homologous recombinant arms VTT-TKR of vaccinia virus Tiantan strain TK genes. Luciferase reporter genes and EGFP green fluorescent protein marker genes, which are expressed through promoters P 7.5, are inserted between the upstream homologous recombinant arms VTT-TKL and the downstream homologous recombinant arms VTT-TKR. The invention further discloses a preparation method of the tracking targeting plasmid of the vaccinia virus Tiantan strain. According to the tracking targeting plasmid of the vaccinia virus Tiantan strain, the targeted removal of the TK genes in the vaccinia virus Tiantan strain can be achieved, and a recombinant Tiantan strain virus with the TK genes removed is prepared.

Description

technical field [0001] The invention belongs to the technical field of biological genetic engineering, and relates to a tracer targeting plasmid of the Tiantan strain of vaccinia virus, and also relates to a preparation method of the tracer targeting plasmid of the Tiantan strain of vaccinia virus. Background technique [0002] Vaccinia virus Tian Tan (VTT) strain is a unique vaccine strain in my country. In order to improve the oncolytic efficacy of vaccinia virus Tian Tan strain VTT, the corresponding targeting plasmid is needed. Thymidine kinase TK can provide a high level of nucleotide pool for viral DNA replication, and the level of TK in normal cells is low. Removing the viral TK gene makes the virus more inclined to replicate in tumor cells and cannot replicate in normal cells earlier, so Removing the TK gene of VTT improves the tumor selectivity of the virus. At present, targeting plasmids targeting the Tiantan strain of vaccinia virus are mainly used for the preven...

Claims

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Application Information

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IPC IPC(8): C12N15/63C12N15/90C12N15/65C12N15/66
CPCC12N15/63C12N15/902C12N15/65C12N15/66
Inventor 马茜汪洋徐文郭红霞刘梅李怡萱
Owner XIAN MEDICAL UNIV
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