Primer for identifying whether aspergillus flavus strain produces aflatoxin or not and application thereof

A technology of Aspergillus flavus strain and aflatoxin, applied in the biological field, can solve problems such as time-consuming and labor-intensive, and achieve the effects of high reliability, high speed and high specificity

Active Publication Date: 2014-04-16
INST OF AGRO FOOD SCI & TECH CHINESE ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, to determine whether Aspergillus flavus produces toxin is mainly to cultivate the Aspergillus flavus strain first, then extract the toxin, and finally detect the toxin content, which is very time-consuming and laborious

Method used

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  • Primer for identifying whether aspergillus flavus strain produces aflatoxin or not and application thereof
  • Primer for identifying whether aspergillus flavus strain produces aflatoxin or not and application thereof
  • Primer for identifying whether aspergillus flavus strain produces aflatoxin or not and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1. Design and synthesis of primers for identifying whether Aspergillus flavus strains produce aflatoxin

[0037] According to the sequences of the toxin synthesis genes aflR and hypB, two sets of PCR primers were designed, namely aflR-F / aflR-R and hypB-F / hypB-R. The details are shown in Table 1. The primers were synthesized by Shanghai Sangong, and can also be synthesized by conventional primer synthesis methods.

[0038] Table 1 Primer sequences of different toxin synthesis genes

[0039]

Embodiment 2

[0040] Embodiment 2, the identification of whether the Aspergillus flavus strain produces toxin

[0041] Tested strains: Aspergillus flavus strains DW-12, DW-14, XinZ-11, XinZ-27, XinZ-33, YC-19, HG-14, QD-1, JZ-2, GZ- 17, and DW-5, DW-6, XinZ-15, XinZ-24, YC-10, HG-12, HG-24, QD-15, FX-1, GZ-9.

[0042] 1. Strain DNA Extraction

[0043] Use an inoculation needle to scrape 0.2 g of mycelium from the PDA plate, put it into a centrifuge tube, pour liquid nitrogen, and then put it in a high-speed tissue grinder to grind the mycelium into powder. Transfer the ground mycelium powder to a 2mL centrifuge tube, add 750μL of extraction solution (recipe: solvent is water, solute and its concentration: 100mmol / L Tris-HCl, 150mmol / L EDTA, pH 8.0), 50 Stand at ℃ for 2 minutes, then add 150 μL of 10% (10g / 100ml) SDS aqueous solution, mix well, add 450 μL of benzyl chloride, shake the centrifuge tube vigorously to make the mixture in the tube milky. Place in a water bath at 50°C for 1 hou...

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Abstract

The invention discloses a primer for identifying whether an aspergillus flavus strain produces aflatoxin or not and an application thereof. The primer for identifying the aspergillus flavus strain which does not produce the aflatoxin, provided by the invention, comprises the following two primer pairs: a primer pair 1 consisting of two single-chain DNAs (deoxyribonucleic acids) as shown in sequence 1 and sequence 2 in a sequence table and a primer pair 2 consisting of two single-chain DNAs as shown in sequence 3 and sequence 4 in the sequence table. Experiments prove that the primer for identifying whether the aspergillus flavus strain produces toxin or not, provided by the invention, has the characteristics of high sequence specificity, high speed of PCR (polymerase chain reaction) amplification identification, high credibility and the like, provides a great technical means for fast, accurate and high-throughput screening of aspergillus flavus strains which do not produce the toxin, and has very important significance for controlling the pollution of aflatoxin in agricultural products.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a primer for identifying whether aflatoxin is produced by an aspergillus flavus strain and an application thereof. Background technique [0002] Aflatoxins are a class of secondary metabolites mainly produced by fungi such as Aspergillus flavus and Aspergillus parasiticus. In my country, aflatoxins are mainly produced by Aspergillus flavus. Aflatoxin has the "three causes" of carcinogenicity, teratogenicity, and cell mutation. Only a dose of 0.294 mg / kg can cause acute poisoning death in sensitive animals. Its main target is the liver, which is the primary cause of malignant tumors. One of the main factors of hepatocellular carcinoma (Hepatocellular carcinoma), it can lead to liver necrosis and canceration in as short as 24 weeks, and can also cause acute lesions of the kidney and adrenal gland. Therefore, effective prevention and control of aflatoxin pollution is of great significanc...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/04C12N15/11C12R1/67
CPCC12Q1/6895C12Q2600/16
Inventor 周露刘阳魏丹丹张初署邢福国赵月菊王龑
Owner INST OF AGRO FOOD SCI & TECH CHINESE ACADEMY OF AGRI SCI
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