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TK (Thymidine Kinase) gene removed recombinant VTT (Tian Tan strain) oncolytic VACA (Vaccinia Virus) and preparation and application thereof

A technology of vaccinia virus and gene, which is applied in the field of preparation of recombinant Tiantan strain oncolytic vaccinia virus, can solve the problem of lack of removal and achieve the effect of reducing virus toxicity and improving tumor selectivity

Pending Publication Date: 2019-05-28
西安彤盛生物科技有限公司
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

At present, there is still a lack of oncolytic virus of the Tiantan strain that removes the TK gene and has the ability to track in vivo

Method used

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  • TK (Thymidine Kinase) gene removed recombinant VTT (Tian Tan strain) oncolytic VACA (Vaccinia Virus) and preparation and application thereof
  • TK (Thymidine Kinase) gene removed recombinant VTT (Tian Tan strain) oncolytic VACA (Vaccinia Virus) and preparation and application thereof
  • TK (Thymidine Kinase) gene removed recombinant VTT (Tian Tan strain) oncolytic VACA (Vaccinia Virus) and preparation and application thereof

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[0037] -

[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 DNA sequence of the wild vaccinia virus Tiantan strain in the present invention is the sequence of GenBank No. AF095689.1 (VRL14-FEB-2000).

[0042] The Tiantan strain of vaccinia virus in the following examples is preserved in the Molecular Virus and Viral Immunology Laboratory of Xi'an Medical College (this laboratory, the same below). The pMV-VTTTK-ELuc plasmid is preserved by our laboratory; cell lines CV-1 and HuTK...

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Abstract

The invention discloses a TK (Thymidine Kinase) gene removed recombinant VTT (Tian Tan strain) oncolytic VACA (Vaccinia Virus). A VTT oncolytic VACA is subjected to recombination transformation, a TKgene is removed, and an EGFP (Enhanced Green Fluorescent Protein) gene and a Luciferase gene are expressed. The invention further discloses a preparation method of the TK gene removed recombinant VTToncolytic VACA, which comprises the steps of: recombining a wild type VTT virus and a targeting plasmid aiming at the TK gene of the VTT virus in cells to obtain virus suspension; screening and purifying the virus suspension to obtain the attenuated VTT oncolytic VACA. According to the TK gene removed recombinant VTT oncolytic VACA, which is disclosed by the invention, the TK gene is used as an attenuation target, the EGFP gene and the Luciferase gene are expressed, virus virulence is reduced, and tumor selectivity of the virus is improved; the TK gene removed recombinant VTT oncolytic VACA has a function of monitoring distribution of virus particles in a body, provides a novel antitumor drug for the clinic, can be used for researching and developing a live vaccine and is applicable to various tumors such as the lung cancer, the liver cancer, the melanoma and the like.

Description

technical field [0001] The invention belongs to the technical field of biogenetic engineering, and relates to a recombinant Tiantan strain oncolytic vaccinia virus with TK gene removed, and also relates to a preparation method and application of the recombinant Tiantan strain oncolytic vaccinia virus with TK gene removed. Background technique [0002] Tumor biotherapy technology is a brand-new tumor treatment method that has emerged in recent years. It has the characteristics of small side effects, high anti-tumor activity, and wide adaptability. [0003] As an oncolytic virus, vaccinia virus (VACV) has natural tropism for tumors, can selectively infect tumor cells and replicate in them, and will not be quickly cleared by the immune system. Vaccinia virus Tiantan strain (TianTan strain, VTT) is a unique strain of vaccinia virus in my country. In order to further improve the selectivity of VTT to tumor cells, some modification of VTT is needed. Thymidine kinase (TK) provides...

Claims

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Application Information

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IPC IPC(8): C12N7/01C12N15/63C12N15/90C12N15/65A61P35/00
Inventor 马茜汪洋
Owner 西安彤盛生物科技有限公司
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