Primer pair and reagent box for identifying acremonium terricola mutagenic strain MKL18

A technology of Acremonium terrestris and mutagenic strains, which is applied in the field of bacterial species identification, and can solve the problems of leakage and leakage, inability to distinguish, and inability to monitor and identify well.

Active Publication Date: 2015-01-28
合肥迈可罗生物工程有限公司
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Problems solved by technology

[0007] However, the Acremonium acremonium mutagenic strain MKL18 did not have any difference in colony and microscopic morphology compared with the wild origin strain, and could not be distinguished by naked eye or microscopic observation
Although the applicant filed an invention patent application for the strain and preserved the strain in the depository institution, it will not be able to monitor and identify this high-quality strain if it is leaked due to internal reasons in the production company.

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  • Primer pair and reagent box for identifying acremonium terricola mutagenic strain MKL18
  • Primer pair and reagent box for identifying acremonium terricola mutagenic strain MKL18
  • Primer pair and reagent box for identifying acremonium terricola mutagenic strain MKL18

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Embodiment Construction

[0033] The present invention will be described in further detail below in conjunction with the accompanying drawings and specific embodiments.

[0034] In this embodiment, a method for identifying acremonium acremonium mutant strain MKL18 comprises the following steps: 1. cultivation of acremonium acremonium and extraction of chromosomal DNA

[0035] ①Cultivation of Acremonium acremonium: Take the wild starting strain of Acremonium acremonium, the mutant strain of Acremonium acremonium MKL18 and the 20th generation strain stored in a -80°C ultra-low temperature refrigerator (Chinese invention patent CN 201410016617.2) , inoculated in 10ml of liquid Chapei medium (NaNO 3 0.2%, K 2 HPO 4 0.2%, KCl 0.5%, MgSO 4 0.4%, FeSO 4 0.001%, sucrose 2%, and the rest is water) in an Erlenmeyer shaker flask, 25° C., 200 rpm shaking culture for 96 hours.

[0036]②Chromosomal DNA extraction: take 3ml of bacterial liquid, centrifuge at 5000rpm for 5 minutes to obtain mycelia, suspend and w...

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Abstract

The invention discloses a primer pair and a reagent box for identifying an acremonium terricola mutagenic strain MKL18. A base sequence of the primer pair is as follows: forward primer: 5'-CCAATGCCCTTCGGGGCTCTCTGG-3', and reverse primer: 5'-GGGATTGGGTAATTTGCGCGCCT-3'. The invention discovers that variation occurs in 18S rDNA of the acremonium terricola mutagenic strain MKL18, and the variation can stably inherit in a subculture strain. On this basis, the primer pair disclosed by the invention is designed for a sequence which is 230bp long in acremonium 18S rDNA and is relatively conservative, and the primer pair is used for identifying the acremonium terricola mutagenic strain MKL18 and a subculture strain thereof, so as to specifically separate the acremonium terricola mutagenic strain MKL18 from a wild original strain thereof.

Description

technical field [0001] The invention belongs to the technical field of strain identification, and in particular relates to a primer pair and a kit for identifying acremonium acremonium mutagenic strain MKL18. Background technique [0002] Cordyceps sinensis is a kind of precious traditional Chinese medicinal material, which is widely used in traditional Chinese medicine and modern clinical medicine. Wild Cordyceps has harsh requirements on its growth conditions, resulting in low yields. Therefore, it is widely seen to use its fungal anamorphs for industrial submerged fermentation to produce various medicines, nutritional health products and feed additives. Through excavating abundant wild resources and artificial mutagenesis in the laboratory, various strains with high-quality production performance have also been widely reported. [0003] Since the isolation and identification methods of the anamorphs of Cordyceps fungi are not very systematic and standardized, the identif...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/04C12N15/11C12R1/645
CPCC12Q1/6895C12Q2600/156C12Q2600/166
Inventor 石应辉胡青松桂向东李晓祥
Owner 合肥迈可罗生物工程有限公司
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