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Microporous cell for evaluating bacteria chemotaxis and evaluation method thereof

A chemotaxis and bacterial technology, applied in sterilization methods, microorganism-based methods, biochemical equipment and methods, etc., can solve the problems of cumbersome and time-consuming operations, and achieve simple operation, simple and easy-to-implement, good The effect of chemotaxis

Inactive Publication Date: 2014-11-26
RES INST OF SHAANXI YANCHANG PETROLEUM GRP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These methods have their own characteristics: the bacterial surge plate method can observe the chemotaxis of bacteria under macroscopic conditions, but it needs to prepare medium and aseptic operation, which makes the operation cumbersome and time-consuming; capillary method and agarose inner plug method Both can reflect the chemotaxis of bacteria to a certain primer under microscopic conditions, but both are limited by microscopic conditions

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  • Microporous cell for evaluating bacteria chemotaxis and evaluation method thereof
  • Microporous cell for evaluating bacteria chemotaxis and evaluation method thereof
  • Microporous cell for evaluating bacteria chemotaxis and evaluation method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0043] Microporous chamber filter method to study the principle of bacterial chemotaxis.

[0044] The invention can reflect the chemotaxis ability of bacteria to different primers. Since the non-water-soluble primers cannot form a concentration gradient in the water phase, the present invention is only applicable to the research on the chemotaxis of bacteria to the water-soluble primers. The microporous chamber in the present invention is used to study the chemotaxis of bacteria, and the chemotaxis experiment in the microporous chamber is designed according to the fact that bacteria can actively migrate through a filter membrane with a certain pore size. The structure of the microporous chamber is as figure 1 shown. The formation of the primer concentration gradient is realized through the PVPF polycarbonate filter membrane. Bacteria can pass through the PVPF polycarbonate filter membrane with a certain pore size along the concentration gradient, and the bacterial penetratio...

Embodiment 2

[0050] Effects of filters with different pore sizes on bacterial chemotaxis.

[0051] Take Escherichia coli as the experimental strain, add 200ul 10mM sucrose as a primer in the lower chamber of the microporous chamber, cover the lower chamber with 0.22um, 3um, 5um, and 8um filter membranes, install the upper chamber on the filter membrane, and place the upper chamber on the filter membrane. The bacterial concentration OD is 0.6 in the chamber (the bacterial concentration is about 10 8 CFU) 800ul of bacteria in the late logarithmic period, place the small chamber at 37°C to incubate the bacteria for 30 minutes, then take it out to measure the number of bacteria entering the lower chamber, calculate the penetration rate of the bacteria according to the formula, repeat the experiment many times, the results are shown in Table 1 Show:

[0052]

Embodiment 3

[0054] Chemotaxis of bacteria to different primers.

[0055] Take Escherichia coli as the experimental strain, add 200ul 10mM sucrose, buffer, and distilled water as primers in the lower chamber of the microporous chamber, cover the lower chamber with a 3um pore size filter membrane and install the upper chamber on it, and add The bacterial concentration OD is 0.6 (the bacterial concentration is about 10 8 CFU) 800ul of bacteria in the late logarithmic period, place the small chamber at 37°C for 30 minutes, take it out and measure the number of bacteria entering the lower chamber, calculate the penetration rate of the bacteria according to the formula, and repeat the experiment many times. The results are shown in Table 2.

[0056]

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Abstract

The invention relates to a microporous cell for evaluating bacteria chemotaxis and an evaluation method thereof. The microporous cell for evaluating bacteria chemotaxis comprises an upper cell for placing bacteria liquid and a lower cell for placing a primer, wherein a filter membrane is arranged between the upper cell and the lower cell. The bacteria chemotaxis evaluation method of the microporous cell comprises the following steps of: 1), taking bacteria, washing the bacteria, detecting the concentration of the bacteria; 2) adding a primer into a lower cell, covering the lower cell with the filter membrane, mounting the upper cell on the filter membrane, and adding bacteria liquid in the upper cell; 3) placing the cell under the optimal temperature for bacterial growth to enter the bacteria smoothly into the lower cell via the filter membrane, after culturing for a period of time, taking out the cell; 4) taking out the bacteria liquid in the upper cell and the lower cell to detect the concentration of the bacteria, counting the amount of bacteria entering into the lower layer, calculating the penetration rate of the bacteria, and reflecting the chemotaxis of the bacteria to the primer through the penetration rate. The microporous cell is simpler to operate, is free from preparation of culture medium and sterile operation, can be reused for many times, and needs shorter detection time.

Description

technical field [0001] The invention relates to a microporous chamber for evaluating bacterial chemotaxis and an evaluation method thereof. Background technique [0002] The behavior of motile microorganisms in response to the concentration gradient of various chemical substances is called chemotaxis. Microbial chemotaxis is of great interest in research in areas such as situ bioremediation, biofilm formation, pathogenesis of infection, migration of microorganisms in subsurface environments and soils, and microbial oil recovery. [0003] At present, there are three main methods for studying bacterial chemotaxis: bacterial surge plate method, capillary method, and agarose plug method. These methods have their own characteristics: the bacterial surge plate method can observe the chemotaxis of bacteria under macroscopic conditions, but it needs to prepare medium and aseptic operation, which makes the operation cumbersome and time-consuming; capillary method and agarose inner p...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12M1/12C12Q1/68C12Q1/04C12R1/01
Inventor 梁凯强王香增高瑞民江绍静洪玲李世强赵丽薛媛
Owner RES INST OF SHAANXI YANCHANG PETROLEUM GRP