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Use of saccharomyces cerevisiae ERG4 mutants for expressing mammalian glucose transporters

A technology for glucose transport and Saccharomyces cerevisiae, applied in the field of yeast strains, can solve the problems of retention, inactive transporters, and low transporter activity

Inactive Publication Date: 2010-05-12
SANOFI AVENTIS DEUT GMBH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, some difficulties have arisen that appear to be associated with these systems: the activity of heterologously expressed transporters is too low, the system's own glucose transporter, a significant proportion of the transporter is retained intracellularly or even produces an inactive transporter. protein

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0092] Use of Yeast Strains

[0093] All yeast strains described here were derived from strain CEN-PK2-1C (MATa leu2-3, 112ura3-52trp1-289his3-Δ1MAL2-8 C SUC2). The preparation of yeast strains with deletion of the hexose transporter (HXT) gene has been described by Wieczorke et al. (FEBS Lett. 464, 123-128 (1999): EBY-18ga(MATaΔhxt1-17Δgal2Δagt1Δstl1 leu2-3, 112ura3-52trp1-289 his3 -Δ1MAL2-8 C SUC2), EBY.VW4000 (MATa Δhxt1-17Δgal2Δagt1Δmph2Δmph3Δstl1 leu2-3, 112ura3-52trp1-289his3-Δ1 MAL2-8 C SUC2)). The medium is based on 1% yeast extract and 2% peptone (YP), while the minimal medium consists of 0.67% Difco Yeast Nitrogen Base (YNB) without amino acids, and contains additives required for co-nutrients and different carbon source. Yeast cells were grown on a rotary shaker or agar plates at 30°C under aerobic conditions. By measuring the optical density at 600nm (OD 600 ) or measure the diameter of yeast colonies to monitor cell growth.

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Abstract

The invention relates to yeast strains in which a human GLUT4 transporter or a human GLUT1 transporter can be functionally expressed and in particular GLUT4 transport proteins which can be particularly easily functionally expressed in yeast strains.

Description

field of invention [0001] The present invention relates to yeast strains capable of functionally expressing human GLut4 and GLut1 transporters. Background technique [0002] Most heterotrophic cells transport glucose into the cell via specific transport proteins. Different organisms have developed different mechanisms for mediating glucose transport, such as especially the proton co-transport system, Na + Glucose transporters, binding protein-dependent systems, phosphotransferase systems, and systems for facilitated diffusion. In eukaryotes, the family of glucose transporters encoded by the GLUT gene (GLUT = glucose transporter) in mammals and by the HXT gene (HXT = hexose transporter) in Saccharomyces cerevisiae Diffused glucose uptake. The transporters belong to the larger family of sugar transporters. They are characterized by the presence of 12 transmembrane helices and multiple conserved amino acid residues. Glucose transport plays an important role in disorders as...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/12C07K14/62C12Q1/02C12Q1/54C12N1/19C12P21/02A61P3/10C12R1/865C12R1/85A61K38/00
CPCC07K14/62A61K38/00A61P3/10C12N15/11C12N1/18
Inventor G·米勒S·德卢盖伊D·福斯E·博尔斯
Owner SANOFI AVENTIS DEUT GMBH
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