Target plasmid for removing Tian Tan strain VGF gene and preparation method thereof

A technology of VTT-VGFR and VTT-VGFL, which is applied in the field of preparation of fluorescently labeled targeting plasmids and targeting plasmids

Pending Publication Date: 2019-05-14
XIAN MEDICAL UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In order to remove the VGF gene in VTT, we need a targeting plasmid for the VGF gen

Method used

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  • Target plasmid for removing Tian Tan strain VGF gene and preparation method thereof
  • Target plasmid for removing Tian Tan strain VGF gene and preparation method thereof
  • Target plasmid for removing Tian Tan strain VGF gene and preparation method thereof

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Embodiment Construction

[0045] The present invention will be described in detail below in conjunction with the accompanying drawings and specific embodiments.

[0046] 1. Main reagents and materials:

[0047] 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).

[0048] 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). The pMV-PRO plasmid and p-EGFP-Luc plasmid are preserved by our laboratory.

[0049] Second, the synthesis of the targeting plasmid that removes the VGF gene of the Tiantan strain:

[0050]The construction process of the targeting plasmid pMV-VTTVGFE used to remove the VGF gene of the vaccinia virus Tiantan strain in the present invention: the 7325-8179th position of the DNA sequence of the wild vaccinia virus Tiantan strain (that is,...

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Abstract

The invention discloses a target plasmid for removing a Tian Tan strain VGF gene. The target plasmid comprises an upstream homologous recombination arm VTT-VGFL and a downstream homologous recombination arm VTT-VGFR of a vaccinia virus Tian Tan strain VGF gene, wherein a fluorescent-labeled gene EGFP is inserted between the upstream homologous recombination arm VTT-VGFL and the downstream homologous recombination arm VTT-VGFR, the fluorescent-labeled gene EGFP is started to express by a promoter P7.5, and a nucleotide sequence of the fluorescent-labeled gene EGFP is shown as sequence 4. The invention also discloses a preparation method of the target plasmid for removing the Tian Tan strain VGF gene. The target plasmid for removing the Tian Tan strain VGF gene can remove the VGF gene from the vaccinia virus Tian Tan strain so as to prepare an attenuated vaccinia virus Tian Tan strain with the VGF gene removed.

Description

technical field [0001] The invention belongs to the technical field of biological genetic engineering, and relates to a targeting plasmid for removing the VGF gene of the Tiantan strain, specifically a fluorescently labeled targeting plasmid for removing the VGF gene of the Tiantan strain, and also relates to the targeting plasmid for removing the VGF gene of the Tiantan strain Preparation. 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 study and improve the oncolytic efficacy of VTT, it...

Claims

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Application Information

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IPC IPC(8): C12N15/63C12N15/90C12N15/65C12N15/66
Inventor 马茜汪洋姚佳李浩辛邦温阳
Owner XIAN MEDICAL UNIV
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