HSV1-TK (herpes simplex virus type 1-thymidine kinase) gene recombination oncolytic adenovirus, preparation method and application thereof

A HSV1-TK, 1.HSV1-TK technology, applied in the field of recombinant oncolytic adenovirus, can solve problems such as difficulty in achieving transfection efficiency, achieve good development and application prospects, simple preparation method, and good stability

Inactive Publication Date: 2010-07-07
ARMY MEDICAL UNIV
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Problems solved by technology

In order to ensure the safety of in vivo treatment, traditional viral vectors use replication-defective viral vectors, but it is technically difficult to achieve 100% transfection efficiency

Method used

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  • HSV1-TK (herpes simplex virus type 1-thymidine kinase) gene recombination oncolytic adenovirus, preparation method and application thereof
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  • HSV1-TK (herpes simplex virus type 1-thymidine kinase) gene recombination oncolytic adenovirus, preparation method and application thereof

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Embodiment Construction

[0026] Hereinafter, preferred embodiments of the present invention will be described in detail with reference to the accompanying drawings. The experimental method that does not indicate specific conditions in the preferred embodiment is usually according to conventional conditions, such as described in the Molecular Cloning Experiment Guide (Third Edition, J. Sambrook et al., translated by Huang Peitang, etc., Science Press, 2002) conditions, or as recommended by the manufacturer.

[0027] 1. Preparation of recombinant oncolytic adenovirus with HSV1-TK gene

[0028] 1. Construction of HSV1-TK gene recombinant expression vector

[0029] Herpes simplex virus I (HSV1) was cultured overnight in LB liquid medium at a temperature of 37°C and a shaking speed of 250r / min, and the genome was extracted with a genomic DNA isolation and purification kit (Shanghai Sangon Bioengineering Co., Ltd.) DNA, according to the kit instructions.

[0030] According to the HSV1-TK gene sequence of...

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Abstract

The invention discloses an HSV1-TK (herpes simplex virus type 1-thymidine kinase) gene recombination oncolytic adenovirus, a preparation method and an application thereof. The genome of the recombination oncolytic adenovirus sequentially comprises a promoter, an adenovirus E1 area gene and an HSV1-TK full length coding gene from upstream to downstream. The recombination oncolytic adenovirus effectively combines the characteristics of a suicide gene and an oncolytic virus and ensures that the oncolytic virus is limited in a tumor cell and efficiently replicated, and meanwhile, an expression product of the HSV1-TK gene converts a nontoxic prodrug into a cytotoxic substance to cause the condition that the tumor cell suicides. The recombination oncolytic adenovirus has the advantages of high transfection efficiency, good stability, wide tumor treatment spectrum, simple preparation method, and the like, can be used for preparing anti-tumor medicaments and has good development and application prospects in the field of tumor gene treatment.

Description

technical field [0001] The invention relates to a recombinant oncolytic adenovirus, in particular to a recombinant oncolytic adenovirus with HSV1-TK gene, and to a preparation method and application of the recombinant oncolytic adenovirus. Background technique [0002] Tumor is an important disease that threatens human health, and the research on new methods of tumor treatment has always been the focus of medical and life science research. Tumor biotherapy has been recognized by the world medical community as the fourth major treatment method after surgery, radiotherapy and chemotherapy, among which immunotherapy and gene therapy are the most effective and cutting-edge scientific research projects in biotherapy. [0003] The principle of tumor gene therapy is to introduce the target gene into target cells with gene transfer technology, make it express the gene to obtain specific functions, and then execute or mediate the killing and inhibition of tumors, so as to achieve the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/01C12N15/85A61K35/76A61K48/00A61P35/00C12R1/93A61K35/761
Inventor 杨曌吴玉章
Owner ARMY MEDICAL UNIV
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