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Common expression vector between 2 subtype receptor of human somatotropin release in hibiting factor and cytosine deaminease of colibacillus, establishment and application

A technology of Escherichia coli cytosine deaminase and co-expression vector is applied in the direction of introducing foreign genetic material, gene therapy, anti-tumor drugs, etc. by using vector, which can solve the problem that gene therapy is difficult to achieve satisfactory clinical therapeutic effect, restrict progress, and Difficulty positioning and treating problems

Inactive Publication Date: 2005-01-12
汪静
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Problems solved by technology

However, at present, it seems difficult for a single gene therapy to achieve a satisfactory clinical therapeutic effect, and it is difficult to locate the treatment of transgenes in the experiment, and it is difficult to monitor the curative effect, which limits the progress of research.

Method used

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  • Common expression vector between 2 subtype receptor of human somatotropin release in hibiting factor and cytosine deaminease of colibacillus, establishment and application
  • Common expression vector between 2 subtype receptor of human somatotropin release in hibiting factor and cytosine deaminease of colibacillus, establishment and application
  • Common expression vector between 2 subtype receptor of human somatotropin release in hibiting factor and cytosine deaminease of colibacillus, establishment and application

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Embodiment Construction

[0031] The human somatostatin receptor subtype 2 (SSTR-2) and Escherichia coli cytosine deaminase (CD) co-expression vector has the sequence listed in Sequence Listing 1. Its construction method is carried out according to the following steps:

[0032] (1) RT-PCR cloning of human SSTR2 gene

[0033] Culture cells containing human SSTR2 gene, such as human embryonic kidney 293 (HEK293) cells, small cell lung cancer cell line NCI-H446, until the cells grow to about 8×10 6 , discard the culture medium, add 1ml TRI ZOL According to the operating instructions, total cellular RNA was extracted and dissolved in DEPC-treated H 2 O, apply SuperScript TM The kit is reverse-transcribed into the first strand of cDNA, using str5 and str3 as primers, PCR amplifies human SSTR2 cDNA, and the result of electrophoresis of the PCR product shows that a DNA fragment of the expected size (1110bp) appears, see figure 2 Electrophoresis photos of hSSTR2 gene obtained by RT-PCR: left band. DL2000...

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Abstract

This invention relates to a common expression vector of human somatostatin receptor subtype 2 and coli bacillus cytosine dearminizing enzyme and its set-up and application. Internal ribosome enter site (IRES) is used to connect in series with SSTR-2 and CD to let them express simultaneously to set up new report gene system, which combines with the cancer multiplication suppression mediated by somatostatin receptor, radiation killing, image display and CD to jointly kill cancers.

Description

technical field [0001] The invention relates to molecular cloning and gene therapy in the field of biotechnology, in particular to the construction of a co-expression vector of human somatostatin receptor 2 subtype and Escherichia coli cytosine deaminase and its establishment and application. Multiple effects such as imaging and internal irradiation kill tumor cells. Background technique [0002] Tumor is one of the most serious diseases that threaten human health in today's society. In clinical practice, surgery, radiotherapy, chemotherapy and other methods are used for treatment. However, in most cases, these treatment methods cannot fundamentally cure the disease, and have toxic side effects. Large, so the efficacy is not satisfactory. The emergence of gene therapy has brought new hope to people. The so-called gene therapy is to transfer a certain nucleic acid substance (gene expression carrier or other) into the patient's body to realize the expression of a specific gen...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K48/00A61P35/00C12N15/70
Inventor 汪静贾林涛王喆李国权
Owner 汪静
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