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Immune tbid gene and its coded protein and application

A gene coding and gene technology, applied in the application field of medicine, can solve the problems of poor prognosis of breast cancer, no effective treatment drugs and methods, etc., and achieve the effect of less toxic side effects, direct killing effect and high efficiency

Active Publication Date: 2007-05-30
FOURTH MILITARY MEDICAL UNIVERSITY
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  • Claims
  • Application Information

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Problems solved by technology

[0003] HER-2 is an epidermal growth factor receptor encoded by a proto-oncogene, which is widely and high-level expressed in tumor cells such as breast cancer and ovarian cancer. It is a recognized tumor marker, and many studies have shown that, The prognosis of HER-2-positive breast cancer is often very poor, and the tumor shows a strong ability to metastasize. At present, there are no effective drugs and methods for treatment

Method used

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  • Immune tbid gene and its coded protein and application
  • Immune tbid gene and its coded protein and application
  • Immune tbid gene and its coded protein and application

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Embodiment Construction

[0030] 1. Construction of immune tbid gene

[0031] The immune tbid gene constructed by the present invention includes three parts, which are respectively the single-chain antibody (e23sFv) of anti-tumor surface antigen HER-2, the transmembrane domain of Pseudomonas aeruginosa exotoxin (PEA) with transmembrane function and the transmembrane domain with inducible cell Truncated Bid (tBid) for apoptotic function.

[0032] (1) Cloning of truncated bid gene (tbid)

[0033] ① Cloning of the full-length bid gene

[0034] Total RNA was extracted from human lymphoma Jurkat cells, and cDNA was synthesized by reverse transcription. Add 2 μL of reverse transcription product, 5 μL of 10×PCR buffer, and 50 mM MgCl in sequence in a thin-walled tube dedicated to PCR 2 1.5 μL, 1 μL of upstream and downstream primers (1E, 2XS), 1 μL of 10 mM dNTP, 0.5 μL of Taq DNA polymerase and H 2 O 38 μL for PCR reaction. The PCR reaction conditions were: denaturation at 96°C for 30 seconds; annealin...

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Abstract

The invention supplies an immunity tbid gene, the albumen coded by the gene, and the application in medicament manufactured by the gene and the albumen in tumor curing. The invention builds secretion type immunity target gene and after secreting the gene would produce effector molecule that could kill tumor cell and have the function of immunity and watching. The immunity tbid gene has safety, high efficiency, and persistence. It could be used as a new method to cure HER-2 masculine tumor.

Description

technical field [0001] The present invention relates to gene therapy in the field of biotechnology, in particular to genetic engineering methods for constructing an immune tbid gene (we named it Immuno-tbid), the protein encoded by the above-mentioned gene, and the preparation of the gene and protein for the treatment of tumors. in the application. Background technique [0002] With the continuous emergence of new medical technologies, the surgical treatment of primary tumors, especially those diagnosed early, has been significantly improved. However, when tumor cells metastasize to distant organs, especially those with extensive metastases, conventional treatments such as chemotherapy, radiotherapy, and surgery cannot achieve the goal of completely controlling the tumor. Secreting and expressing targeted pro-apoptotic proteins, inducing tumor cell apoptosis and killing tumors, this idea will guide the emergence of some new tumor gene therapy programs with high efficiency a...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/13C07K16/18A61K38/16A61K39/00A61P35/00
Inventor 杨安钢裘秀春许彦鸣贾林涛于翠娟赵晶张立红王立锋孟艳玲王智范清宇王成济
Owner FOURTH MILITARY MEDICAL UNIVERSITY
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