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Bacterial capable of over-expressing membrane protein, and application thereof

A technology of overexpression and membrane protein, applied in bacteria, using vectors to introduce foreign genetic material, microorganisms, etc., can solve the problems of membrane protein hydrophobicity, cell decapacity toxicity, etc.

Inactive Publication Date: 2014-08-20
DONGHUA UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Combined with the difficulty of purification and crystallization of membrane proteins, the heterologous expression of membrane proteins has also become a bottleneck in the research of membrane proteins due to problems such as hydrophobic properties, limited cell decapacity, and toxicity to host cells.
Especially for the prokaryotic expression of eukaryotic membrane proteins, the incompatibility of the host often brings greater difficulties: as of 2005, there is no report of a membrane protein that obtains structural information through recombinant expression

Method used

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  • Bacterial capable of over-expressing membrane protein, and application thereof
  • Bacterial capable of over-expressing membrane protein, and application thereof
  • Bacterial capable of over-expressing membrane protein, and application thereof

Examples

Experimental program
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Embodiment 1

[0018] Source of bacteria: This strain comes from oil-contaminated soil. The specific isolation method is as follows:

[0019] (1) Enrichment: Take 1g of oil-contaminated soil and put it in the enrichment medium (0.5g of beef extract, 1g of peptone, 0.5g of sodium chloride, pH7.0-7.6), and incubate at 28°C and 120 rpm for 48h . Then take 2 mL of its culture solution and transfer it to another enrichment medium, and further enrich and cultivate for 48 hours at 28°C and 120 rpm.

[0020] (2) Preliminary screening: Take 100 μL oil plate culture medium of the final enrichment solution (crude oil 5g, ammonium chloride 6g, magnesium sulfate 0.2g, calcium chloride 0.001g potassium dihydrogen phosphate 4.1g, disodium hydrogen phosphate 14g, EDTA0. 001g, agar 20g, yeast powder 0.5g, H 2 O1000ml, pH7.4), a single colony with a large transparent circle was initially screened out.

[0021] (3) Re-screening: Spot the strains obtained from the primary screening on the phosphoric acid dig...

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Abstract

The present invention relates to bacterial capable of over-expressing membrane protein, and an application thereof, wherein the bacterial is Escherichia coli, is named Escherichia coli 1098-3, and has the preservation number of CGMCC No.8696, and the 16S rRNA sequence is represented by SEQ ID NO:1. According to the present invention, the bacterial has great tolerance on membrane protein, the membrane protein can be subjected to over-expression by using the bacterial, and the good application prospects are provided.

Description

technical field [0001] The invention belongs to the field of biosurfactants, in particular to a bacterium capable of overexpressing membrane protein and its application. Background technique [0002] Research on E. coli in the 1960s and 1970s made it the most commonly recognized organism in nature. Escherichia coli has two remarkable features: simple operation and high-density culture in cheap medium. These features and more than ten years of experience in foreign gene expression make it the most efficient expression of heterologous protein in most scientific research applications. The most commonly used prokaryotic expression system. Despite E. coli's many advantages, not every gene is efficiently expressed in it. [0003] The natural expression level of membrane proteins is low under normal physiological conditions, and their heterologous mass expression becomes a prerequisite for functional and structural studies. Combined with the difficulties in the purification and ...

Claims

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Application Information

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IPC IPC(8): C12N1/21C12N15/63C12R1/19
Inventor 张中鸽曹张军
Owner DONGHUA UNIV