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Molecular chaperone plasmid system and application thereof

A technology of molecular chaperones and plasmids, which is applied in the field of genetic engineering and can solve problems such as poor foreign proteins

Pending Publication Date: 2021-08-03
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, in the study of exogenous protein expression using Corynebacterium glutamicum, there is no good solution for some exogenous proteins existing in inclusion bodies, so it is urgent to develop the endogenous molecular chaperone plasmid system of Corynebacterium glutamicum and integrate it Applied to foreign protein expression

Method used

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  • Molecular chaperone plasmid system and application thereof
  • Molecular chaperone plasmid system and application thereof
  • Molecular chaperone plasmid system and application thereof

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Experimental program
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Embodiment 1

[0035] The Corynebacterium glutamicum strain expressing scFv (pXMJ19-scFv) was taken out from the -80°C refrigerator, and after being streaked on a solid plate for activation, the bacteria were picked and inoculated into 10mL LBB medium, 200r / min, 30°C, and cultured for 12h. Transfer to EPO medium to prepare competent cells. Electrotransform p99E-GSl1 / GSL2 / DJ1EK / DJ2EK and pEC-XK99E into the above-mentioned competent cells. Spread on the LBB plate containing 30mg / L kanamycin and 10mg / L chloramphenicol and culture at 30°C for 24h, pick a single colony into 10mL LBB liquid medium, add 30mg / L chloramphenicol and 30mg / L card Namycin, 200r / min, 30°C, cultivated for 12h. According to the 2% inoculum amount, the above-mentioned bacterial solution was transferred to 5 bottles of LBB containing 10mL, 30mg / L chloramphenicol and 30mg / L kanamycin liquid medium, and the inducer IPTG (isopropyl- β-D-thiogalactoside), 200r / min, 30°C, cultivated for 24h.

[0036] Collect the above-mentioned...

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Abstract

The invention discloses a molecular chaperone plasmid system and application thereof, the molecular chaperone plasmid system comprises four expression vectors, and the four expression vectors are GSL1, GSL2, DJ1EK and DJ2EK. The molecular chaperone plasmid system can effectively promote soluble expression of corynebacterium glutamicum foreign protein.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to a molecular chaperone plasmid system and its application. Background technique [0002] Corynebacterium glutamicum is a commonly used strain for the industrial production of industrial products such as amino acids and higher alcohols, and belongs to Gram-positive bacteria. In recent years, due to the discovery that Corynebacterium glutamicum does not produce endotoxin and has no extracellular hydrolase activity, it is often used for the expression of exogenous recombinant proteins. At present, the methods for improving the expression of foreign proteins by using the expression system of Corynebacterium glutamicum mainly include the screening of expression vectors and elements, host transformation and medium optimization. [0003] Chaperones are a class of proteins that help other peptide chains to fold correctly but are not present in the target protein. Studies hav...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/77C12N1/21C12R1/15
CPCC12N15/77C07K14/00
Inventor 刘秀霞王亚丽崔思楠白仲虎杨艳坤李业刘春立
Owner JIANGNAN UNIV
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