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Escherichia coli anti-tumor targeted engineering strain, and building method and application thereof

A technology of Escherichia coli and engineering strains, applied in anti-tumor drugs, applications, genetic engineering and other directions, can solve the problems of body infection treatment effect, difficult expression of eukaryotic anti-tumor proteins, poor economy and other problems, achieve high-efficiency expression, good The effect of antitumor efficacy

Active Publication Date: 2018-01-12
HUNAN NORMAL UNIVERSITY
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  • Abstract
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Problems solved by technology

[0004] There are certain defects in bacterial treatment of tumors: when live bacteria are used to directly treat patients, due to the poor pathogenicity and targeting of some bacteria, it may cause serious infection or poor therapeutic effect, and the economy is not good.
[0005] Although Escherichia coli EcN is highly targeted to solid tumors, it can specifically

Method used

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  • Escherichia coli anti-tumor targeted engineering strain, and building method and application thereof
  • Escherichia coli anti-tumor targeted engineering strain, and building method and application thereof
  • Escherichia coli anti-tumor targeted engineering strain, and building method and application thereof

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Embodiment Construction

[0047]The present invention will be described in further detail below in conjunction with the accompanying drawings and embodiments.

[0048] (1) Obtaining Tum-5 gene by PCR splicing

[0049] The strains and plasmids used in the present invention are shown in Table 1, and the primers used are shown in Table 2.

[0050] Table 1 Bacterial strains and plasmids used in the present invention

[0051]

[0052]

[0053] Table 2 Primers used in the present invention

[0054]

[0055] First, use Tum 5-M as a template, Tum 5-A-F and Tum 5-A-R as upstream and downstream primers, perform the first round of PCR to obtain a 138bp fragment; then use the 138bp fragment as a template, Tum 5-B-F and Tum 5 -B-R as primers, perform the second round of PCR to obtain a 214 bp fragment; then use the 214 bp fragment as a template, Tum 5-C-F and Tum 5-C-R as primers, perform a third round of PCR to obtain a 258 bp fragment; Finally with this 258bp fragment as template, Tum 5-F-Nco I or Tum...

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Abstract

The invention relates to an escherichia coli anti-tumor targeted engineering strain, and a building method and application thereof. The escherichia coli anti-tumor targeted engineering strain, i.e., Escherichia coli Nissle 1917(Tum-5) is preserved in CCTCC (China Center for Type Culture Collection), and the preservation number is CCTCC NO:M 2017345. The anti-tumor targeted engineering strain contains an anti-tumor secretion express vector pET28a-Pvhb-SUMO-Tum 5; and the soluble expression and the secretory expression of anti-tumor blood vessels of tumstatin can be realized to generate an active region Tum-5. The invention also comprises the building method of the escherichia coli anti-tumor targeted engineering strain, and application of the escherichia coli anti-tumor targeted engineeringstrain to preparation of anti-tumor medicine. The tumor inhibiting rate of the escherichia coli anti-tumor targeted engineering strain on B16-bearing melanoma C57BL/6 mice reaches up to 52.95 percent.

Description

technical field [0001] The invention relates to an Escherichia coli anti-tumor targeting engineering strain, its construction method and its application in the preparation of anti-tumor drugs. Background technique [0002] Bacteria have a long history of use in cancer therapy. Although the clinical mechanism by which bacteria treat tumors is unclear, bacteria play a very important role in the treatment of certain diseases, especially cancer. Traditional chemotherapy and radiotherapy treatments are often unable to completely eliminate tumors, which is attributed to tumor cells in the tumor anaerobic microenvironment in solid tumors. The oxygen concentration in normal tissues of most mammals is In tumor tissue, the normal concentration near the stroma at the border of the necrotic area gradually decreased to the hypoxic state (O 2 <0.02%). A series of studies have shown that tumor-targeted expression of cancer therapeutic genes may be an effective and safe approach in ...

Claims

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Application Information

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IPC IPC(8): C12N1/21C12N15/70C12N15/12A61K48/00A61K38/17A61P35/00C12R1/19
Inventor 夏立秋何恋丁学知孙运军胡胜标
Owner HUNAN NORMAL UNIVERSITY
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