Lactobacillus casei ghost, preparation method and application thereof

A technology of Lactobacillus casei and Lactobacillus, applied in the biological field, can solve the problems of not being able to guarantee 100% complete lysis of bacteria, spreading pathogens and infections, and not being able to be popularized and applied, so as to achieve heavy workload, strengthen immunogenicity, and improve immunity horizontal effect

Inactive Publication Date: 2013-04-03
NORTHEAST AGRICULTURAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

[0003] So far, the bacteria ghosts that have been reported are all prepared by pathogenic bacteria, and the bacteria ghost preparation technology cannot guarantee 100% complete lysis of the bacteri

Method used

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  • Lactobacillus casei ghost, preparation method and application thereof
  • Lactobacillus casei ghost, preparation method and application thereof
  • Lactobacillus casei ghost, preparation method and application thereof

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Embodiment 1

[0039] The preparation of embodiment 1 Lactobacillus casei bacteria shadow and the identification of basic characteristics

[0040] 1. Materials and methods

[0041] 1.1 Test material

[0042] 1.1.1 Plasmids and strains

[0043] Lactobacillus casei L.casei 393 was donated by NIZO Research Institute in the Netherlands, and Escherichia coli competent JM109 and pMD18-T Simple vectors were purchased from Dalian Bao Biological Engineering Co., Ltd.

[0044] 1.1.2 Molecular biology reagents

[0045] Ex Taq DNA polymerase, dNTP and restriction endonuclease were purchased from TaKaRa Company, T4 DNA ligase was purchased from NEB Company, plasmid miniprep kit and DNA gel recovery kit were purchased from Axygen Company.

[0046] 1.1.3 Primer synthesis

[0047] Primers were designed using oligo6.0 software, primer 1 and primer 2 were used to amplify the perforin (hocb) gene fragment from the phage genome; primer 3 and primer 4 were used to amplify the lysin (lycb) gene fragment from ...

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Abstract

The invention discloses a lactobacillus casei ghost, a preparation method and an application thereof, belonging to the biotechnology field. According to the lactobacillus casei ghost, lactobacillus casei is used as a research object, a recombinant vector containing cloned lytic genes is converted into the lactobacillus casei through separation of bacteriophage of the lactobacillus casei, and prediction and cloning of the lytic genes, and then the lytic genes are induced to be expressed to prepare the lactobacillus casei ghost. The lactobacillus casei ghost is used as an antigen delivery carrier safely and reliably in the true sense; besides, muramyl dipeptide in the cell wall of the lactobacillus casei has a good adjuvant effect and can remarkably enhance the immunogenicity of heterogeneous antigen; furthermore, the mass production of the lactobacillus casei ghost can be realized through fermentation, and the lactobacillus casei ghost can be preserved at room temperature for a long time after being frozen and dried, so that the cost for application of production practice is further reduced; and a new idea and a new method for further designing a novel, safe and effective drug, particularly a vaccine delivery system, which are applied to the prevention and treatment of the diseases are provided.

Description

technical field [0001] The invention relates to a method for preparing bacterial ghosts, the bacteria ghosts prepared by the method and applications thereof. In particular, it relates to a method for preparing a Lactobacillus casei ghost and the bacterial ghost prepared by the method. Belongs to the field of biotechnology. Background technique [0002] Bacterial ghosts are empty shells of bacteria formed after the bacterial cell wall is lysed and the cell contents are removed. It has a complete bacterial surface antigen structure and can be directly used as a vaccine. At the same time, the ghost is also a good carrier, and its residual inner membrane structure can be used to load biomacromolecules after resealing. In the past, bacteriophages were prepared by using E protein, a protein expressed by Φx174 bacteriophage, which cleaves the cell wall of Gram-negative bacteria. Construct the gene of E protein into an inducible expression vector, transform Gram-negative bacteri...

Claims

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Application Information

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IPC IPC(8): C12N1/21C12N15/34C12N15/74A61K47/46C12R1/245
Inventor 乔薪瑗李一经唐丽杰葛俊伟兰宇
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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