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A method for promoting the mass expression of glial fibrillary acidic protein in Escherichia coli

An acidic protein, Escherichia coli technology, applied in microorganism-based methods, chemical instruments and methods, biochemical equipment and methods, etc., can solve problems such as protein inactivation, increase purification costs, etc., achieve strong expression activity, and improve inclusion bodies. problems, high-expression effects

Active Publication Date: 2022-06-03
XIAMEN BIOTIME BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

On the one hand, it increases the cost of purification, on the other hand, the purification process may cause protein inactivation

Method used

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  • A method for promoting the mass expression of glial fibrillary acidic protein in Escherichia coli
  • A method for promoting the mass expression of glial fibrillary acidic protein in Escherichia coli
  • A method for promoting the mass expression of glial fibrillary acidic protein in Escherichia coli

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Embodiment

[0036] Embodiment A method for promoting the massive expression of glial fibrillary acidic protein in Escherichia coli

[0037] The method for promoting the massive expression of glial fibrillary acidic protein in Escherichia coli comprises the following steps:

[0038] S1, use the software OptimumGene TM Analyze the gene sequence encoding the GFAP protein, perform codon optimization, and obtain the optimized GFAP gene sequence;

[0039] S2. The optimized GFAP gene sequence obtained in step S1 is introduced into the pGEX-4T-2 plasmid, and the sequence information is verified by sequencing, and the successfully constructed recombinant plasmid pGEX-4T-2-GFAP-opt can be obtained;

[0040] with the software OptimumGene TM The gene sequence SEQ ID NO. 1 encoding GFAP protein was analyzed to find out its codon usage preference and the different sites of E. coli usage preference. For codon sites with different usage preferences, the codons preferred by Escherichia coli were used t...

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Abstract

The invention belongs to the field of biotechnology, and in particular relates to a method for promoting the mass expression of glial fibrillary acidic protein in Escherichia coli. The present invention optimizes the codons of the base sequence of glial fibrillary acidic protein, the optimized sequence information is shown in SEQ ID NO.2, and further transforms the pGEX-4T-2 vector, adding the ompa signal peptide, The signal peptide sequence information is shown in SEQ ID NO.4. Through the method provided by the invention, the secretion of glial fibrillary acidic protein in Escherichia coli is increased, and the expression of glial fibrillary acidic protein inclusion body is improved.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for promoting the massive expression of glial fibrillary acidic protein in Escherichia coli. Background technique [0002] Glial fibrillary acidic protein (GFAP) is a specific cytoskeletal protein unique to the central nervous system, and is the only marker protein expressed by astrocytes. 40% of the total number of human brain cells. GFAP is a marker protein of astrocytes and astrocyte-derived tumors, supports and nourishes neurons, and participates in many biological functions of astrocytes, including cell proliferation and division, maintenance The normal physiological function of the blood-brain barrier, autophagy, maintaining the balance of neurotransmitters, etc. GFAP is an astrocyte-specific intermediate filament protein and a marker protein for gliomas. Under normal conditions, GFAP in the cytoplasm of cells is degraded in circulation, and the level of...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/70C12N15/67C12N15/66C12N15/62C12P21/02C12R1/19
CPCC12N15/70C07K14/47C12N2800/22C07K2319/02Y02A50/30
Inventor 徐玉艳
Owner XIAMEN BIOTIME BIOTECHNOLOGY CO LTD