Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Anti-tumor targeting engineering bacteria and bacterial agent and method for preparing bacterial agent

A technology of anti-tumor and engineering bacteria, which is applied in the direction of anti-tumor drugs, biochemical equipment and methods, and methods based on microorganisms. It can solve the problems of normal tissue toxicity, etc., and achieve the effect of short fermentation cycle and simple growth conditions

Active Publication Date: 2013-01-16
湖南晴天生物科技有限公司
View PDF0 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Anticancer drugs on the market can not only inhibit and kill tumor cells, but also have a great toxic effect on normal tissues of the human body

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Anti-tumor targeting engineering bacteria and bacterial agent and method for preparing bacterial agent
  • Anti-tumor targeting engineering bacteria and bacterial agent and method for preparing bacterial agent
  • Anti-tumor targeting engineering bacteria and bacterial agent and method for preparing bacterial agent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] Example 1: Example of the construction method of the anti-tumor targeting engineered bacteria of the present invention

[0055] PCR amplified P lac 启动子LacI末端与LacZ前端序列GCGCCCAATACGCAAA CCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGTGAGCGCAACGCAATTAATGTGAGTTAGCTCACTCATTAGGCACCCCAGGCTTTACACTTTATGCTTCCGGCTCGTATGTTGTGTGGAATTGTGAGCGGATAACAATTTCACACAGGAAACAGCTATGACCATGATT;设计引物,Plac-F: CACCCTCATCAGTGCCAACATAGTAA GCCAGTATACACTCCGCTAGCGCG CGCCCAATACGCAAACCGCCT , the underlined part is the specific primer, and its front sequence is the homology arm; Plac-R: ATTTCTAGAGGATCC CCGGGTACCGAGCTCGAATTCGTA ATCATGGTCATAGCTGTTTC , the underlined part is the specific primer, its front sequence is the homology arm, the pBlueScript KS(+) plasmid is used as the template, and Plac-F / Plac-R is used as the primer to amplify the 311bp truncated P lac For the promoter and homology arm part, the reaction conditions are: pre-denaturation at 94°C for 4 min; denaturation a...

Embodiment 2

[0065] Example 2: Example of the preparation method of the anti-tumor targeting engineered bacterial agent of the present invention

[0066] Process reference Figure 8 :

[0067] (1) Activation of bacteria

[0068] Streak-inoculate the preserved recombinant Escherichia coli LMA1 on the slant of the solid seed medium, and culture it upside down at 37°C for 12 hours; pick a single colony and streak it, and culture it upside down at 37°C for 12 hours for further purification;

[0069] (2) Preparation of fermented seed liquid

[0070] Pick Escherichia coli LMA1 to inoculate into 10ml containing 12.5μg.mL -1 Chloramphenicol in LB medium, 200r.min at 37°C -1 Shake culture for 8h to mid-logarithmic growth, inoculate 10mL of culture solution into 1000mL containing 12.5μg.mL -1 Chloramphenicol LB medium, 200r.min at 37°C -1 Shake culture for 13 hours to obtain the fermentation liquid of the primary seed tank; sterilize the secondary seed tank at 121°C for 30 minutes, put it in...

experiment example 1

[0086] Experimental Example 1: Inhibitory Effect of Gene Recombined Escherichia coli LMA1 Bacterial Agent on the Growth of B16F10 Mice Melanoma

[0087] The purpose of this experiment is to study the inhibitory effect of Escherichia coli LMA1 bacterial agent on the growth of melanoma in B16F10 mice

[0088] Mouse melanoma B16-F10 cells similar to human melanoma were routinely cultured in 1640 medium with 10% FCS in 5% CO2 cultured at 37°C in a cell culture incubator. The cells in the exponential proliferation phase were digested with 0.25% trypsin, collected by centrifugation, washed 3 times with PBS, stained with trypan blue, and the cell suspension was counted, and the cell viability was detected to be over 95%, and finally resuspended to 1×10 cells in PBS. 6 / mL. Forty C57BL / 6 female mice of 6-8 weeks were selected, and 100ul of B16F10 mouse melanoma cells were subcutaneously injected on the right side of the back of the mice. After 6 days, the mice were randomly divided ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
densityaaaaaaaaaa
Login to View More

Abstract

The invention provides anti-tumor targeting engineering bacteria, a bacterial agent and a method for preparing the bacterial agent. The anti-tumor targeting engineering bacteria contain an anti-tumor secretory expression vector pLAC-HA, and anti-tumor targeting engineering bacteria are named Escherichia coli LMA1 through classification. The integration expression vector used for construction is pLAC, and the cut Plac constitutive expression promoter is used to reinforce the expression of small peptides Ehp. The invention also discloses the method for preparing an anti-tumor targeting engineering bacterial agent. Animal test results show that the anti-tumor targeting engineering bacteria have the quite high targeting property on tumors, but do not have obvious adverse influence on normal lab mice, and the bacterial agent prepared by the anti-tumor targeting engineering bacteria is a medicament for effectively treating tumors.

Description

technical field [0001] The invention relates to an anti-tumor targeting engineering bacterium, a bacterial agent and a preparation method of the bacterial agent, in particular to an Escherichia coli Nissle 1917 containing and secreting and expressing an anti-tumor AZURIN protein, a bacterial agent and a preparation method of the bacterial agent. Background technique [0002] Cancer incidence and mortality continue to rise as the world's population and longevity increase. According to the "World Cancer Report" provided by the World Health Organization (WHO), according to the current trend of cancer incidence, the incidence of cancer worldwide will increase by 50% in 2020, and the number of new cancer patients worldwide will reach 15 million each year. At present, the cancer with the highest incidence rate in the world is lung cancer, with 1.2 million new patients every year; followed by breast cancer, with about 1 million new patients every year; followed by intestinal cancer...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/21C12N15/09A61K35/74A61P35/02C12R1/19A61K35/741
Inventor 夏立秋丁学知张允雷胡胜标
Owner 湖南晴天生物科技有限公司
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com