Lactic acid bacteria having ampicillin resistance, and preparation and applications of preparation of lactic acid bacteria having ampicillin resistance

A technology of ampicillin and resistance, which is applied in the field of preparation and application of lactic acid bacteria and its preparations, and can solve problems such as the lack of systemic resistance genes of lactic acid bacteria

Inactive Publication Date: 2015-12-23
JINAN UNIVERSITY
View PDF2 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, human research on antibiotic resistance is mostly focused on pathogenic bacteria, and there is a lack of systematic research on the antibiotic resistance of lactic acid bacteria and whether the resistance gene can be tr...

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
  • Lactic acid bacteria having ampicillin resistance, and preparation and applications of preparation of lactic acid bacteria having ampicillin resistance
  • Lactic acid bacteria having ampicillin resistance, and preparation and applications of preparation of lactic acid bacteria having ampicillin resistance
  • Lactic acid bacteria having ampicillin resistance, and preparation and applications of preparation of lactic acid bacteria having ampicillin resistance

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Example 1 Preliminary Screening of Probiotics

[0050] The specific operation steps of strain isolation (including culture temperature, time, etc.) feature description)

[0051] 1. Culture medium:

[0052] Lactobacillus MRS separation solid medium: Accurately weigh 10.0g of beef extract peptone, 10.0g of tryptone, 5.0g of yeast extract, 5.0g of anhydrous sodium acetate (NaAc), 2.0g of triammonium citrate, K 2 HPO 4 2.0g, MgSO 4 ·7H 2 O0.58g, MnSO 4 ·H 2 O0.28g, glucose 15.0g, 1.0mL Tween-80 into the beaker, distilled water and stir evenly, add 15.0g agar and stir, after the drug is completely dissolved, add distilled water to 1000mL, monitor and adjust the pH of the medium to 6.2~ with a pH meter 6.4, sterilize with damp heat at 121°C for 15 minutes, and set aside.

[0053] Bifidobacterium improved MRS separation solid medium: accurately weigh 10.0g of beef extract peptone, 10.0g of tryptone, 5.0g of yeast extract, 15.0g of glucose, 5.0g of sodium acetate, 2.0g ...

Embodiment 2

[0090] The biological activity evaluation of embodiment 2 probiotics

[0091] The bacteria to be tested were Bifidobacterium longum HB25 and Lactobacillus acidophilus HL9 obtained in Example 1, and the control strain was Lactobacillus rhamnosus GG (LGG for short, ATCC53103). The bacterial strains frozen in the laboratory at -80°C were taken respectively and thawed in a water bath at 37°C. Take 100 μL of each bacteria in 100 mL of lactic acid bacteria MRS enrichment broth in a biological safety cabinet for activation and recovery. Shake the bacteria at 200rpm on an air-bath shaker at 37°C for 12h, centrifuge the bacteria solution at 4000rpm for 15min, remove the supernatant after centrifugation, wash the bacterial pellet with sterile saline for 3 times, and then transfer the bacterial pellet to the standard turbidimetric solution In the same test tube as the centrifuge tube, use sterile physiological saline to dilute to the concentration that needs to be higher than the standar...

Embodiment 3

[0128] 1. Analysis of strain plasmids

[0129] According to Wan Qian's method (Wan Qian, Cao Jian, Wu Xingquan, etc. Optimization of the extraction method of natural plasmids of lactic acid bacteria [J]. China Biotechnology Journal, 2010,30(2):94-99.) Carry out plasmid extraction, take Centrifuge 5-10 mL of the strain to be tested overnight, discard the supernatant, resuspend with 250 μL of solution P1 (add lysozyme, to a final concentration of 20 mg / mL), and bathe in water at 37°C for 30 minutes; add 250 μL of solution P2 and mix quickly, then add 350 μL Solution P3, mix well, centrifuge at 12000rpm for 10min; take the supernatant and add it to the spin column, centrifuge at 9000rpm for 1min, discard the liquid in the centrifuge tube; wash the spincolumn twice with 500μL WashBuffer, put the spincolumn into a new EP tube and centrifuge at 12000rpm for 2min , and then add 50 μL LutionBuffer (preheated at 60° C.), leave it for 2 minutes, centrifuge, and set aside. Detected by 1...

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

No PUM Login to view more

Abstract

The present invention discloses lactic acid bacteria having ampicillin resistance, and preparation and applications of a preparation of the lactic acid bacteria having ampicillin resistance. According to the present invention, human manure is separated under antibiotic pressure to obtain two strains of lactic acid bacteria have drug resistance, an antibiotic sensitivity test is performed, and whether plasmid DNA exists is detected; PCR amplification is performed to analyze the drug resistance gene localization conditions of the strains and explore the molecular mechanisms of the antibiotic resistance of the lactic acid bacteria, and safety evaluation is performed on the screened lactic acid bacteria having the resistance gene; whether the screened resistance strain provides an adjustment effect on intestinal tract imbalance is verified, and an antibiotic type intestinal tract flora imbalance model is used so as to provide potential treatment methods for diarrhea, flora imbalance and other diseases caused by the use of antibiotics in the clinic; and according to the obtained strain of the present invention, the intestinal tract probiotics can survive under the antibiotic pressure when the antibiotic is clinically used again while the ordinary probiotics can not survive, such that the treatment method can be provided for the diarrhea caused by the antibiotics in the clinic.

Description

technical field [0001] The invention belongs to the field of biomedicine, in particular to the preparation and application of ampicillin-resistant lactic acid bacteria and preparations thereof. Background technique [0002] There are a large number of microorganisms living in the human body, the main ones are bacteria, as we know, they can settle in the human intestinal tract and form complex groups or micro-ecosystems. According to statistics, there are 1014 bacteria living in the intestinal tract of healthy adults, which is 10 to 20 times the total number of human cells, and there are as many as nearly 500 species of bacteria. Intestinal bacteria are the largest and most important part of the human body. The bacteria count in the stool reaches 10 12 . The body's host cells, gut microbes, and other microbes constitute a complex system that interacts to form a variety of biological effects. [0003] Intestinal probiotics (Probiotics) are a type of intestinal bacteria that...

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
IPC IPC(8): C12N1/20A61K35/747A61K35/745A61P1/00C12R1/01C12R1/23
Inventor 孙晗笑田鹏杨妍利时雨丁清韦丕金刘照兵黄俊丽
Owner JINAN UNIVERSITY
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap