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Method for regulating chromosome genome functions by using combined promoter

A promoter, combinatorial technology, used in combinatorial chemistry, introduction of foreign genetic material using vectors, recombinant DNA technology, etc.

Inactive Publication Date: 2010-11-24
苏州神洲基因有限公司
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  • Summary
  • Abstract
  • Description
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Problems solved by technology

[0007] However, there are certain defects in the above methods
These methods can only study one gene at a time, or are limited to one regulatory unit

Method used

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  • Method for regulating chromosome genome functions by using combined promoter
  • Method for regulating chromosome genome functions by using combined promoter

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Embodiment Construction

[0036] The technical scheme of the present invention will be further described below in conjunction with specific examples. The term "microorganism" refers to prokaryotic organisms including bacteria and eukaryotic microorganisms including yeast, fungi and higher organisms.

[0037] The present invention involves a method for simultaneous assembly of multiple DNA fragments. In order to achieve this goal, the DNA fragments to be ligated are designed as follows: For each pair of fragments that need to be ligated, the distal region of one DNA fragment needs to contain regions homologous to the proximal region of the other DNA fragment. In order to promote the connection of DNA fragments in a predetermined direction and order, the distal and proximal regions of each set of selected recognition sequences are unique (different from the recognition sequence regions of other DNA fragment pairs). This method can connect a large number of DNA fragments. At the same time, the present inve...

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Abstract

The invention provides a method for regulating chromosome genome functions by using a combined promoter, comprising the following step of: regulating and optimizing the expression of a plurality of genes in a related metabolic pathway by constructing an artificially combined type promoter sublibrary to replace a self promoter on chromosome, and therefore, the selected microorganisms can effectively generate needed products. The combined promoter comprises four artificially combined double-stranded DNA units with homologous ends, wherein the four units are assembled by a selectively marked and reformed regulating and controlling sublibrary, a reformed core promoter sublibrary and a reformed ribosome bind site library and are transformed into one or a plurality of target cells to replace the self promoters of a plurality of target chromosome genes in a specific cell pathway to cause high-expression target genes of a large amount of target cells. The method can be used for constructing a plurality of integrated bacterial strains in multiple-chromosome sites. The artificially combined type promoter can accurately, simultaneously or singly regulate and control several kinds of gene activity in cells and is suitable for metabolism optimization and metabolism control analysis in eukaryotic cells and procaryotic cells.

Description

Technical field [0001] The present invention relates to a method for regulating the function of chromosome genome by using a combined promoter, which regulates and optimizes the expression of multiple genes in related metabolic pathways by constructing an artificial combined promoter library and replacing the self-promoters on the chromosome with it , So that the selected microorganism can effectively produce the desired product, which belongs to the technical field of microbial genetic modification. Background technique [0002] Microorganisms (such as Escherichia coil) are widely used as industrial reactors for the large-scale production of chemicals, enzymes and active pharmaceutical ingredients, agricultural chemicals and biofuels. However, strains that exist in nature rarely have excellent performance under ideal conditions. Therefore, the biggest challenge in the production of these chemicals using biological methods is how to optimize cell metabolism in order to reduce pr...

Claims

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Application Information

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IPC IPC(8): C12N15/63C12N15/113C40B50/06
Inventor 王德明
Owner 苏州神洲基因有限公司
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