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Functionalized bacterium magnetic particles for expressing specific recombinant proteins and preparation method thereof

A bacterial magnetic particle and recombinant protein technology, applied in the fields of molecular biology and nano-magnetic bead materials, can solve the problems of poor binding specificity and low loading of IgG, and achieve the effect of overcoming mutual influence.

Pending Publication Date: 2016-06-01
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

However, there are generally technical problems in the prior art that the particles formed by combining recombinant proteins with magnetic beads or magnetic particles have poor binding specificity to IgG, and the loading capacity of IgG is low.

Method used

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  • Functionalized bacterium magnetic particles for expressing specific recombinant proteins and preparation method thereof
  • Functionalized bacterium magnetic particles for expressing specific recombinant proteins and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1 The functionalized bacterial magnetic particles expressing specific recombinant protein A

[0045] The structural substance of the functionalized bacterial magnetic particle is formed by fusion expression of specific recombinant protein A through the polypeptide chain and bacterial magnetic particle membrane protein; the amino acid residue of the polypeptide chain is (Ala m GlySer) n (Gly a Ser) b (GlySerAla m ) n , m=3, n=3, a=3, b=5, the sequence is 5-GCAGCCGCTGGAAGCGCAGCCGCTGGAAGCGCAGCCGCTGGAAGCGGAGGTGGAAGCGGAGGTGGAAGCGGAGGTGGAAGCGGAGGTGGAAGCGGAGGTGGAAGCGGAAGCGCAGCCGCTGGAAGCGCAGCCGCTGGAAGCGCAGCCGCT-3;

[0046] Bacterial magnetic particle membrane protein gene mamC;

[0047] 其中,特定重组蛋白A在重组蛋白A的B结构域的氨基酸的基础上,将第29位Gly突变成Ala,序列为5-GCCGACAATAAATTCAACAAAGAACAACAAAATGCGTTCTATGAAATTTTACATTTACCTAATTTAACTGAAGAACAACGTAATGGCTTCATCCAAAGCCTTAAAGACGATCCTTCAGTGAGCAAAGAAATTTTAGCCGAAGCGAAAAAGCTAAACGATGCCCAAGCACCAAAAGGTTCAGGCGGAAGCGGCGGCGGAAGCGGAGGCTCAGGTAGCGCCGACAATAAATTCAA...

Embodiment 2

[0048] Example 2 The functionalized bacterial magnetic particles expressing specific recombinant protein G

[0049] The structural substance of the functionalized bacterial magnetic particle is formed by fusion expression of specific recombinant protein G through the polypeptide chain and bacterial magnetic particle membrane protein; the amino acid residue of the polypeptide chain is (Ala m GlySer) n (Gly a Ser) b (GlySerAla m ) n , m=3, n=3, a=3, b=5, the sequence is 5-GCAGCCGCTGGAAGCGCAGCCGCTGGAAGCGCAGCCGCTGGAAGCGGAGGTGGAAGCGGAGGTGGAAGCGGAGGTGGAAGCGGAGGTGGAAGCGGAGGTGGAAGCGGAAGCGCAGCCGCTGGAAGCGCAGCCGCTGGAAGCGCAGCCGCT-3;

[0050] The bacterial magnetic particle membrane protein gene is mamF;

[0051] 其中,特定重组蛋白G为重组蛋白G的C结构域二聚体,序列为:5-ACTTACAAACTTGTCATTAATGGTAAAACATTGAAAGGCGAAACAACTACTAAAGCAGTCGACGCAGAAACTGCAGAAAAAGCCTTCAAACAATACGCTAACGACAACGGTGTCGATGGTGTGTGGACTTATGATGATGCGACTAAGACCTTTACGGTCACTGAAGGTTCAGGCGGAAGCGGCGGCGGAAGCGGAGGCTCAGGTAGCGGCACTTACAAACTTGTCATTAATGGTAAAACATTGAA...

Embodiment 3

[0052] Example 3 The functionalized bacterial magnetic particles expressing specific recombinant protein A

[0053] The difference between the functionalized bacterial magnetic particles and Example 1 or Example 2 is that the functionalized bacterial magnetic particles are synthesized by recombinant magnetotactic bacteria, and the recombinant magnetotactic bacteria are constructed by the following method:

[0054] a) Constructing a mutant strain of magnetotactic bacteria MSR-1 with deletion of the bacterial magnetic particle membrane protein gene;

[0055] b) Constructing a functionalized bacteria magnetic particle membrane protein fusion expression plasmid;

[0056] c) The fusion expression plasmid of the functionalized bacterial magnetic particle is transformed into the mutant strain of the magnetotactic bacteria MSR-1 prepared in step a to construct the recombinant strain of the magnetotactic bacteria MSR-1.

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Abstract

The invention provides functionalized bacterium magnetic particles for expressing specific recombinant proteins. The bacterium magnetic particle membrane protein is fused with a flexible polypeptide chain to express the specific recombinant proteins. The specific recombinant proteins are Z structure field dimers improved on the basis of recombinant protein A or C structure field dimers improved on the basis of recombinant protein G, and the gene sequences are particularly modified, so that the prepared functionalized bacterium magnetic particles have excellent IgG combining capacity.

Description

technical field [0001] The invention relates to the fields of molecular biology and nano magnetic bead materials, in particular to functionalized bacterial magnetic particles expressing specific recombinant proteins and a preparation method thereof. Background technique [0002] Bacterial magnetic particles are a kind of magnetic nanoparticles produced by magnetotactic bacteria, and the core is Fe 3 o 4 The crystal is coated with a layer of phospholipid biofilm outside, and the particle size is between 30-120nm. The bacterial magnetic particles produced by the same magnetotactic bacteria have basically the same particle size and crystal form, uniform magnetic properties, natural biofilm coating, and good water-soluble and colloidal properties. In addition, because the bacterial magnetic particles are of biological origin, they have good biocompatibility. There are a large number of amino groups on the bacterial magnetic particle membrane, which can be connected with diffe...

Claims

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Application Information

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IPC IPC(8): C12N1/21C12N15/74C12N15/03
CPCC12N15/03C12N15/74C07K14/195C07K14/31C07K14/315
Inventor 张金菊王红光
Owner BEIJING GUO KE RONG ZHI BIOTECH CO LTD
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