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A kind of exogenous promoter suitable for Corynebacterium glutamicum and its application

A technology of Corynebacterium glutamicum and promoter, applied in the fields of genetic engineering and metabolic engineering, can solve the problems such as the inability to meet the growing demand for exogenous proteins

Active Publication Date: 2020-05-22
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] For Corynebacterium glutamicum, people are still limited to the use of limited promoters such as cspB, trc, tac, etc. Among them, the expression level of exogenous proteins in Corynebacterium glutamicum using the tac promoter is still high, but it still cannot satisfy people's needs. Growing demand for exogenous proteins

Method used

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  • A kind of exogenous promoter suitable for Corynebacterium glutamicum and its application
  • A kind of exogenous promoter suitable for Corynebacterium glutamicum and its application
  • A kind of exogenous promoter suitable for Corynebacterium glutamicum and its application

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Embodiment Construction

[0014] The bacterial strains used in the present invention: Bacillus subtilis 168, Corynebacterium glutamicum ATCC 13032 and Escherichia coli DH5α were all purchased from the Chinese University Microbial Resources Data Platform.

[0015] restriction endonuclease Hin d III, Bam H I, Eco R V, T4 DNA ligase, etc. were purchased from Thermo Company; Bsa Type I endonuclease and Q5 ultra-fidelity polymerase were purchased from NEB Company; plasmid extraction kits, PCR product purification kits, and gel recovery kits were purchased from Axygen Company; genome extraction kits were purchased from Tiangen Company; Infusion recombination Kits were purchased from Clontech; antibiotics such as chloramphenicol and kanamycin were purchased from Shanghai Bioengineering Co., Ltd.; other reagents were domestic or imported analytically pure. High-speed refrigerated centrifuge and PCR thermal cycler, ThermoFisher Company, USA; Synergy H4 microplate reader, BioTek Company, USA; Constant te...

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Abstract

The invention provides an exogenous promoter for corynebacterium glutamicum. According to the technical scheme of the invention, the problem in the prior art that the heterologous protein of corynebacterium glutamicum is low in expression level can be solved. The invention also provides the application of the exogenous promoter. The nucleotide sequence of the exogenous promoter for corynebacterium glutamicum is shown as SEQ ID NO: 2, or a complementary nucleotide sequence thereof.

Description

technical field [0001] The invention relates to the technical fields of genetic engineering and metabolic engineering, in particular to an exogenous promoter suitable for Corynebacterium glutamicum and application thereof. Background technique [0002] Corynebacterium glutamicum is a non-pathogenic Gram-positive bacterium, which is widely used in industrial production of amino acids, organic acids, higher alcohols and other industrial products; at the same time, Corynebacterium glutamicum has no endotoxin and no extracellular hydrolysis Enzyme activity, which can secrete correctly folded biologically active functional proteins, so Corynebacterium glutamicum is also a very favorable host for expressing foreign proteins. [0003] A promoter is an important DNA sequence that controls the initiation of transcription of a gene, located upstream of the transcription initiation site. The mRNA transcription level of a gene is one of the main factors affecting the final protein expr...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/113C12N15/77C12N1/21C12P21/02C12R1/15
CPCC07K14/34C12N15/113C12N15/77C12P21/02
Inventor 刘秀霞赵子豪白仲虎杨艳坤戴晓峰
Owner JIANGNAN UNIV
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