Bacterial magnetic particles with recombinant protein G expression capability and application thereof

A technology of bacterial magnetic particles and recombinant proteins, which is applied in the application of nano magnetic beads and medical testing, can solve the problems of increasing the cost of anti-human ball tests, and achieve the effects of improving stability, improving amplification ability, and enhancing load capacity

Active Publication Date: 2016-05-11
BEIJING GUO KE RONG ZHI BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In automated detection, the anti-human globulin test needs to be used in combination with micro-column gel cards. As the cost of micro-column gel cards b

Method used

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  • Bacterial magnetic particles with recombinant protein G expression capability and application thereof
  • Bacterial magnetic particles with recombinant protein G expression capability and application thereof
  • Bacterial magnetic particles with recombinant protein G expression capability and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Example 1 Bacterial magnetic particles expressing recombinant protein G

[0049] A bacterial magnetic particle expressing recombinant protein G, the bacterial magnetic particle expressing recombinant protein G is obtained by expanding, culturing, separating and purifying the recombinant strain of magnetotactic bacteria MSR-1.

Embodiment 2

[0050] Example 2 Bacterial magnetic particles expressing recombinant protein G

[0051] A bacterial magnetic particle expressing recombinant protein G, the bacterial magnetic particle expressing recombinant protein G is obtained by expanding, culturing, isolating and purifying a recombinant strain of magnetotactic bacteria MSR-1;

[0052] The sequence of the recombinant protein G is:

[0053] The magnetotactic bacteria MSR-1 recombinant strain is constructed by the following method:

[0054] a. Construction of a MSR-1 mutant strain of magnetotactic bacteria with deletion of the bacterial magnetic particle membrane protein mamC / mamF gene;

[0055] b. Construction of bacterial magnetic particle membrane protein fusion expression plasmid;

[0056] c. Transfer the functionalized bacterial magnetic particle fusion expression plasmid into the magnetotactic bacteria MSR-1 mutant strain obtained in step a by conjugation to construct the magnetotactic bacteria MSR-1 recombinant s...

Embodiment 3

[0057] Example 3 Bacterial magnetic particles expressing recombinant protein G

[0058] A bacterial magnetic particle expressing recombinant protein G, the bacterial magnetic particle expressing recombinant protein G is obtained by expanding, culturing, isolating and purifying a recombinant strain of magnetotactic bacteria MSR-1;

[0059] The sequence of the recombinant protein G is:

[0060] The preparation method of the bacterial magnetic particles expressing recombinant protein G is as follows:

[0061] a. Construction of a MSR-1 mutant strain of magnetotactic bacteria with deletion of the bacterial magnetic particle membrane protein mamC / mamF gene;

[0062] (a) Amplification of bacterial magnetic particle membrane protein mamC / mamF gene with two homologous fragments on the left and right sides of 700-1000 bp in length; both left and right homologous fragments have two restriction sites;

[0063] (b) The left and right homologous fragments are respectively double dige...

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PUM

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Abstract

The invention provides bacterial magnetic particles with recombinant protein G expression capability. The bacterial magnetic particle with recombinant protein G expression capability are obtained by performing enlarged culture and separation and purification on magnetotactic bacteria MSR-1 recombinant strains. The invention further provides application of the bacterial magnetic particles with recombinant protein G expression capability in detection of blood type irregular antibody IgG. By use of the magnetic property of the bacterial magnetic particles and the effect of an applied magnetic field, the bacterial magnetic particles with recombinant protein G expression capability, provided by the invention, are capable of assisting agglutination of sensitized erythrocytes, so that on the one hand, the usage amount of the bacterial magnetic particles is reduced to the greatest extent, and on the other hand, the detection sensitivity of low-potency IgG antibodies is improved to the greatest extent.

Description

technical field [0001] The invention relates to the fields of nanometer magnetic bead application and medical detection, in particular to bacterial magnetic particles expressing recombinant protein G and application thereof. Background technique [0002] Bacterial magnetic particles are magnetic nanoparticles produced by magnetotactic bacteria, the core of which is Fe 3 O 4 The crystals are coated with a layer of phospholipid biofilm, and the particle size is between 30-120nm. The bacterial magnetic particles produced by the same magnetotactic bacteria are basically the same in particle size and crystal form, have uniform magnetic properties, are coated with natural biofilms, and have good water-soluble and colloidal properties. In addition, because the bacterial magnetic particles are of biological origin, they have better biocompatibility. The bacterial magnetic particle membrane has a large number of amino groups, which can be linked to different functional macromolecu...

Claims

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Application Information

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IPC IPC(8): G01N33/80G01N33/68G01N33/554C12N1/21C12N15/74
CPCC07K14/315C12N15/74G01N33/554G01N33/6854G01N33/80G01N2333/315
Inventor 张金菊王红光
Owner BEIJING GUO KE RONG ZHI BIOTECH CO LTD
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