Genetic transformation selection marker of Pleurotus eryngii

A technology of genetic transformation and screening markers, applied in the field of molecular biology, can solve problems such as the safety and efficiency of Pleurotus eryngii screening markers, and achieve good market application prospects and high transformation efficiency

Inactive Publication Date: 2018-12-07
SHANGHAI ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] The technical problem to be solved by the present invention is to provide a screening marker for genetic transformation of Pleurotus eryngii, which solves the safety and efficiency problems of screening markers for Pleurotus eryngii in the prior art

Method used

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  • Genetic transformation selection marker of Pleurotus eryngii
  • Genetic transformation selection marker of Pleurotus eryngii
  • Genetic transformation selection marker of Pleurotus eryngii

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Embodiment 1

[0020] 1. The method for obtaining the screening marker for Pleurotus eryngii genetic transformation is as follows:

[0021] (1) CTAB method to extract whole genome DNA of Pleurotus eryngii monokaryotic hyphae;

[0022] (2) DNA quality was detected by agarose electrophoresis, and the concentration was detected by Nanodrop;

[0023] (3) obtain the DNA fragment of 2570bp from Pleurotus eryngii monokaryotic hyphae DNA by in vitro amplification technology;

[0024] (4) Mutating the adenine nucleotide at position 2237 into a thymine nucleotide by point mutation technology;

[0025] (5) connecting the mutated DNA fragment into the transformation vector;

[0026] (6) Transform Pleurotus eryngii by PEG-mediated or Agrobacterium-mediated method, and add 2ug / ml wiltin to the medium for screening, and the successfully transformed progeny can grow normally on the wilted screening medium, wild type cannot grow.

[0027] 2. Obtain the Pleurotus eryngii transformant (such as figure 2 s...

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Abstract

The invention relates to a genetic transformation selection marker of Pleurotus eryngii. The selection marker is obtained by a single base mutation of a succinate dehydrogenase iron sulfur subunit gene encoded by the genome of Pleurotus eryngii. The carboxin resistance selection marker obtained by the invention is derived from a single base mutation of a self-gene of Pleurotus eryngii with no introduction of foreign DNA of other species, and has high safety. Meanwhile, the selection marker has high transformation efficiency and can replace a hygromycin resistance screening marker commonly usedin the genetic transformation of Pleurotus eryngii. The selection marker obtained by the invention can be safely and efficiently applied to the genetic improvement of Pleurotus eryngii, lays a foundation for cultivating new varieties of Pleurotus eryngii which accords with the production demand and consumer preferences, and has a good market application prospect.

Description

technical field [0001] The invention belongs to the field of molecular biology, in particular to a screening marker for genetic transformation of Pleurotus eryngii. Background technique [0002] Pleurotus eryngii, also known as Pleurotus eryngii, belongs to the Phylum Fungi, Basidiomycetes, Agaricaceae, Pleurotaceae, and Pleurotus. Pleurotus eryngii has thick flesh, crisp and tender texture, and has an almond-like aroma and abalone-like taste. It is suitable for fresh-keeping and processing, and is a popular edible fungus among consumers. In recent years, the industrialized cultivation of Pleurotus eryngii in my country has developed rapidly, and has become the fastest-growing edible fungus species in facility cultivation after Flammulina velutipes. Utilizing modern molecular biology techniques to improve Pleurotus eryngii varieties to obtain strains with high yield, short growth cycle and strong resistance to miscellaneous bacteria is an important topic in the genetic rese...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/156
Inventor 尚俊军鲍大鹏李燕李焱杨瑞恒汪滢茅文俊唐利华
Owner SHANGHAI ACAD OF AGRI SCI
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