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High efficiency probiotic and preparation method thereof

A high-efficiency, high-expression technology, applied in biochemical equipment and methods, microorganism-based methods, chemicals for biological control, etc. The effect of high activity, great clinical application value and broad medical application prospects

Inactive Publication Date: 2014-12-03
刘志华
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there are also low-efficiency problems in the use of probiotics, that is, under pathological conditions, it is difficult to maintain a high dose of probiotics in the intestinal tract for a long time

Method used

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  • High efficiency probiotic and preparation method thereof
  • High efficiency probiotic and preparation method thereof
  • High efficiency probiotic and preparation method thereof

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Embodiment Construction

[0025] The present invention will be further described below in conjunction with specific embodiments with reference to the accompanying drawings, so as to better understand the present invention.

[0026] This example provides a high-efficiency probiotic bacterium that highly expresses the I58 gene. A recombinant vector containing the I58 gene is transfected into Lactobacillus to obtain a Lactobacillus that highly expresses the I58 gene.

[0027] The preparation method is specifically:

[0028] 1. Clone the I58 gene

[0029] 1.1 Extraction of genomic NDA

[0030] 1) Preheat the extraction buffer at 65°C;

[0031] 2) Add bacteria (Lactobacillus plantarum CGMCC1258) to the preheated buffer (700μl) and mix well, 65°C, 10min;

[0032] 3) Add phenol / chloroform / isoamyl alcohol (25:24:1), shake, and centrifuge at 12,000rpm for 10min;

[0033] 4) Take the supernatant, add chloroform / isoamyl alcohol (24:1) to shake, and centrifuge at 12,000rpm for 10min;

[0034] 5) Take the supe...

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Abstract

The invention provides a high efficiency probiotic. The high efficiency probiotic can highly express the lactic acid bacillus of I58 gene (IMP501-580). A preparation method of the high efficiency probiotic comprises the following steps: designing an amplimer according to an I58 gene sequence, and cloning an I58 gene fragment through PCR; cloning the obtained I58 gene fragment into an expression vector in order to obtain a recombinant vector; and converting the recombinant vector into competent lactic acid bacillus in order to obtain the high efficiency probiotic. The high efficiency probiotic provided by the invention can highly express the I58 gene, has high adhesion activity, no translocation, and no reaction with antibiotics, so the defects of translocation and no antibiotic resistance of common lactic acid bacilli can be well made up, and the colonization, shift and infection of pathogens are inhibited, thereby the high efficiency probiotic has great clinical application values and has a wide medicine application prospect.

Description

technical field [0001] The invention relates to a high-efficiency bacterium, in particular to a lactobacillus highly expressing the I58 gene and a preparation method thereof. Background technique [0002] Probiotics refer to live bacterial preparations and their metabolites that improve the host's micro-ecological balance and play a beneficial role in improving the host's health level and health status. They are widely found in every corner of the earth and are beneficial to humans and animals. Bacteria, they can be taken directly as food additives to maintain the balance of intestinal flora, and the adhesion and colonization in the intestinal tract can inhibit the damage of pathogenic bacteria, which is of great significance for protecting the integrity of the intestinal mucosa; since the 1990s Since the beginning, all kinds of "probiotics" health care products have become popular all over the world. At the same time, the research on "probiotics" has become a hot research t...

Claims

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Application Information

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IPC IPC(8): C12N1/21C12N15/74A61K35/74A01P1/00C12R1/225C12R1/245C12R1/23C12R1/25C12R1/24C12R1/01
Inventor 刘志华秦环龙汪建平
Owner 刘志华
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