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Immune tbid gene, encoded protein and use thereof

A gene coding and gene technology, applied in applications, gene therapy, genetic engineering, etc., can solve the problems of tumor metastasis, poor prognosis of breast cancer, and lack of therapeutic drugs and means.

Inactive Publication Date: 2005-03-16
杨安钢
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Problems solved by technology

[0003] HER-2 is an epidermal growth factor receptor encoded by a proto-oncogene, which is highly expressed in tumor cells such as breast cancer and ovarian cancer. It is a recognized tumor marker, and many studies have shown that HER-2 positive The prognosis of 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, encoded protein and use thereof
  • Immune tbid gene, encoded protein and use thereof
  • Immune tbid gene, encoded protein and use thereof

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Experimental program
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Embodiment Construction

[0031] 1. Construction of immune tbid gene

[0032] 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.

[0033] (1) Cloning of truncated bid gene (tbid)

[0034] ① Cloning of the full-length bid gene

[0035] 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 concerns a tbid gene, protein coding by tbid gene and uses of said gene and protein in tumor drug preparation. In the invention, single chain antibody e23sFv gene which can idiocraticly identify and bind HER-2 positive tumor cell is used to construct a tbid gene with PEA transmembrane structural domain and active truncated bid gene (tbid)which has strong specificity,high activity ,low immunogenicity,sustainable execution on tumor. The invention constructs secretion type immunological target gene. after gene is expressed and secreted, its can produce effector molecule to kill tumor cell. The gene therapy strategy possessing simultaneously the cell immunity and humoral immunity character has obvious advantages compared with other strategy. Immunological tbid gene the invention constructed is safe, highly effective and continuous effect. The invention is hoped to provide a novel method for HER-2 tumor target therapy.

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 tumor control. Secreted expression of targeting protein induces tumor cell apoptosis and kills tumors, which will guide the emergence of some new tumor gene therapy programs with high efficiency and low toxicity. [0003] HER-2 is an epidermal growth...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/17A61K48/00C07K14/435C12N15/12
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 杨安钢
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