Method and element for screening different-intensity ribosome binding sites and promoters

Abstract

The invention belongs to the technical field of zymomonas mobilis, and discloses a method and an element for screening different-intensity ribosome binding sites and promoters. The method comprises the following steps: firstly, screening genes with strong downstream expression according to different genomics data, carrying out Wayne analysis, and screening common gene of the genomics data; predicting the binding site sequence of ribosome with different intensities; acquiring an RBS sequence promoter with specific intensity; acquiring a double-fluorescence reporter gene system framework; acquiring recombinant plasmid; acquiring a strain of the double-fluorescence reporter gene system plasmid containing the promoter with specific RBS intensity; and performing intensity verification. The method can integrate existing systematic biological data, and quickly and efficiently screen promoters with specific intensity. The intensities of the RBS and the promoter which are predicted and screenedby the method have good correlation with experimental data (respectively is R2)0.9 and R2)0.7).

Description

technical field [0001] The invention belongs to the technical field of monocysts, and in particular relates to a method and elements for screening ribosome binding sites and promoters with different strengths. Background technique [0002] At present, the existing technologies commonly used in the industry are as follows: [0003] Zymomonas mobilis is a Gram-negative, facultative anaerobic bacterium with perinatal flagella and a size of 1.4-2.0*4.0-5.0μm; the suitable growth temperature is 30°C, and the pH range it can tolerate Between 3.5 and 9, it can efficiently utilize glucose, fructose and sucrose to produce ethanol through the Enter-Doudoroff pathway (ED pathway). Z.mobilis has many advantages such as high ethanol yield and tolerance, high osmotic pressure tolerance, low biomass, and no need to add oxygen during fermentation, so it has become one of the main production strains of bioethanol. In order to increase the range of its available substrates, improve ethanol ...

AI Technical Summary

Problems solved by technology

[0005] (1) At present, the mining of a large amount of accumulated systems biology data is not enough, so there are few biological components of Z. mobilis that have been identified;

[0006] (2) With the development of metabolic engineering and synthetic biology, the demand for available biological components in Z. mobilis is also increasing, but the number of biological components such as promoters within the genome is less developed, which is not conducive to its Precise metabolic engineering is not convenient for precise regulation in the era of synthetic biology

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