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Amplification primer pair for lactic acid bacteria sequencing, lactic acid bacteria species identification method and application

A technology for amplifying primers and lactic acid bacteria, applied in the field of genetic engineering, can solve problems such as research, unsatisfactory, unfavorable lactic acid bacteria community structure, etc., and achieve good specificity, practicability, and good diversity

Active Publication Date: 2021-09-17
INNER MONGOLIA AGRICULTURAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] At present, the existing lactic acid bacteria primers are mostly used for the identification and quantification of lactic acid bacteria. Conducive to a comprehensive study of the community structure of lactic acid bacteria in the gut

Method used

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  • Amplification primer pair for lactic acid bacteria sequencing, lactic acid bacteria species identification method and application
  • Amplification primer pair for lactic acid bacteria sequencing, lactic acid bacteria species identification method and application
  • Amplification primer pair for lactic acid bacteria sequencing, lactic acid bacteria species identification method and application

Examples

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Embodiment 1

[0035] Use the above-mentioned amplification primers of the present invention to carry out PCR amplification to the lactic acid bacteria of 7 genera and 14 species, specifically comprising the following steps:

[0036] The DNA of 14 strains of 7 genera of common lactic acid bacteria was used as the positive template, and the sterilized and filtered ultrapure water was used as the negative template, and Lab-6F and Lab-6R were used as primers for PCR amplification to obtain PCR amplification products;

[0037] The nucleotide sequences of the primer pairs are as follows:

[0038] Lab-6F: 5'-GCTCAGGAYGAACGCYGG-3', where Y represents C;

[0039] Lab-6R: 5'-CACCGCTACACATGRADTTC-3', where R represents A and D represents T.

[0040] Among them, the 14 bacterial strains are Enterococcus asini (DSM-11492), Enterococcus faecium (ATCC 19434T), Enterococcus Italicus (DSM-15952), Lactobacillus Casei (ATCC-334), Lactobacillus Salivarius (DSM-20555), Lactobacillus Helveticus (1.2278) , Weis...

Embodiment 2

[0047] Compare lactic acid bacteria amplification primer Lab-6 (Lab-6F and Lab-6R) of the present invention and bacterial universal primer 27f / 1492r to the amplification result of lactic acid bacteria in the stool sample, specifically comprise the following steps:

[0048] (1): Amplify the stool DNA dilution with the bacterial universal primer (27f / 1492r) and the amplification primer pair provided in S1 (the nucleotide sequence of the primer pair is the same as in Example 1), and perform PCR amplification System and PCR amplification reaction procedure are all identical with embodiment 1, obtain PCR amplification product;

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Abstract

The invention discloses a pair of amplification primers for sequencing lactic acid bacteria in the intestinal tract. The pair of primers is Lab-6, including Lab-6F and SEQ ID NO.2 whose nucleotide sequence is shown in SEQ ID NO.1 Lab‑6R shown. The amplification primer pair can realize the identification of lactic acid bacteria species, has good specificity and practicability, and can better reflect the diversity of lactic acid bacteria in the intestinal tract. It is suitable for the analysis of lactic acid bacteria in complex samples such as fermented dairy products, feces and soil.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and in particular relates to a pair of amplification primers for sequencing lactic acid bacteria in the intestinal tract, a method for identifying the genus of lactic acid bacteria and its application. This identification method allows the sequencing of lactic acid bacteria in complex samples such as feces. The primer sequence was designed based on the 16S rRNA gene sequence of lactic acid bacteria (mainly including Lactobacillus, Lactococcus, Weissella, Enterococcus, Leuconostococcus, Streptococcus and Pediococcus). The amplified product reaches about 750bp, which meets the sequencing requirements. Background technique [0002] Lactic Acid Bacteria (LAB) is a general term for Gram-positive bacteria that ferment carbohydrates to produce lactic acid, have no spores, do not move or rarely move, and are catalase-negative. Lactic acid bacteria are widespread and play beneficial roles in...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/689C12Q1/6869C12Q1/04C12N15/11
CPCC12Q1/6869C12Q1/689
Inventor 钟智孙志宏张和平白晓晔
Owner INNER MONGOLIA AGRICULTURAL UNIVERSITY
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