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PyrG screening marker recycling method and application

A screening marker, recombinant bacteria technology, applied in the biological field, can solve the problems of repetitive sequence residues, limited genetic screening markers, etc., and achieves the effect of simple operation and wide application range.

Active Publication Date: 2020-12-22
FUJIAN AGRI & FORESTRY UNIV
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Problems solved by technology

[0006] In order to solve the limitation of genetic screening markers in the process of fungal genetic transformation, the impact of different genetic screening markers on the genetic background and traits of fungi, and the traditional URA3 / pyrG The problem of the repetitive sequence residues in the loop-out system, the invention provides a new orotic acid nucleoside-5'-phosphate decarboxylase gene pyrG Methods for Recycling of Genetic Screening Markers

Method used

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  • PyrG screening marker recycling method and application
  • PyrG screening marker recycling method and application
  • PyrG screening marker recycling method and application

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Embodiment Construction

[0030] The inventor established a new orotidine-5'-phosphate decarboxylase gene after careful research pyrG A method for the recycling of genetic selection markers. This method requires only a pyrG The recycling of genetic screening markers can realize multiple genetic transformation operations, avoiding the impact of differences in different genetic screening markers on the study of fungal genetic background and traits, and using this method to eliminate pyrG Genetic screening markers do not need to rely on direct repeats, knockout pyrG There will be no repetitive sequences remaining on the genome after the screening markers, and it will not affect the genetic background and traits of the recombinant bacteria, which can solve the problem of the limitation of genetic screening markers in the process of fungal genetic transformation and the impact of different genetic screening markers on the study of fungal genetic background and traits and traditional URA3 / pyrG The repeat...

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Abstract

The invention discloses a novel PyrG screening marker recycling method and application thereof, and belongs to the technical field of biology. According to the method, the pyrG screening marker can beremoved from a fungal genome without depending on homodromous repetitive sequences at the two ends of the pyrG, redundant repetitive sequence fragments cannot be left on the genome, recombinant bacteria without the pyrG can be used as starting strains again for genetic transformation by using the pyrG, and cyclic utilization of the pyrG screening marker is realized. The pyrG cyclic utilization method disclosed by the invention is suitable for fungi taking the whey glycoside-5'-phosphate decarboxylase coding gene URA3 / pyrG expression element as a genetic selection marker, is simple and easy tooperate, does not influence the genetic background and character of the strain, and has a wide application prospect.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a new pyrG Method and application of screening marker recycling. Background technique [0002] Fungi are a large class of eukaryotic organisms in nature. Some of them provide food for human beings, some become industrial and agricultural strains beneficial to human beings, and some pathogenic strains are harmful to human beings. In the era of functional genomes, the establishment of a genetic manipulation system is inseparable from the research and genetic manipulation of these fungi for the further utilization of beneficial fungi and the prevention and control of fungal diseases. In the process of genetic transformation, the target transformants need to be quickly screened, and the subsequent subculture of the transformants should also avoid the loss of the recombinant gene and maintain the new genetic characteristics obtained by the transgene. All of these must rely on efficient...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/15C12N15/65C12R1/67
CPCC12N9/88C12N15/65C12Y401/01023
Inventor 聂鑫怡王银春薛杨汪世华丁霞飞王佳琪
Owner FUJIAN AGRI & FORESTRY UNIV
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