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Method for promoting mass expression of colloidal fibrillary acidic protein in escherichia coli

A technology based on acidic protein and Escherichia coli, applied in the direction of microorganism-based methods, chemical instruments and methods, biochemical equipment and methods, etc., can solve the problems of increasing purification costs, protein inactivation, etc., and achieve the problem of improving inclusion bodies, strong The effect of increasing expression activity and secretion

Active Publication Date: 2021-06-22
XIAMEN BIOTIME BIOTECHNOLOGY CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

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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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  • Method for promoting mass expression of colloidal fibrillary acidic protein in escherichia coli
  • Method for promoting mass expression of colloidal fibrillary acidic protein in escherichia coli
  • Method for promoting mass expression of colloidal fibrillary acidic protein in escherichia coli

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Embodiment

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

[0037] The method for promoting the mass 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 GFAP protein, perform codon optimization, and obtain the optimized GFAP gene sequence;

[0039] S2. Import the optimized GFAP gene sequence obtained in step S1 into the pGEX-4T-2 plasmid, and verify the sequence information by sequencing to obtain the successfully constructed recombinant plasmid pGEX-4T-2-GFAP-opt;

[0040] OptimumGene TM The gene sequence SEQ ID NO.1 encoding GFAP protein was analyzed to find out its codon usage bias and the sites with different Escherichia coli usage bias. For the codon sites with different usage preferences, the codons preferred by Escherichia coli were used to replace the codon-optimized GFAP protein gene sequ...

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Abstract

The invention belongs to the technical field of biology, and particularly relates to a method for promoting mass expression of colloidal fibrillary acidic protein in escherichia coli. According to the method, codons of a colloidal fibrillary acidic protein base sequence are optimized, the optimized sequence information is shown as SEQ ID NO.2, the pGEX-4T-2 vector is further modified, an ompa signal peptide is added, and the sequence information of the signal peptide is shown as SEQ ID NO.4. According to the method provided by the invention, the secretion of the colloidal fibrillary acidic protein in escherichia coli is increased, and the expression of the colloidal fibrillary acidic protein in an inclusion body form is improved.

Description

technical field [0001] 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. Background technique [0002] Glial fibrillary acidic protein (GFAP) is a specific cytoskeletal protein unique to the central nervous system and a marker protein expressed only by astrocytes. It is widely distributed in various parts of the central nervous system, accounting for about 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 a specific intermediate filament protein of astrocytes and a marker protein of glioma. Under normal c...

Claims

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Application Information

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Patent Type & Authority Applications(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