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Molecular marker for identifying common threewingnut root medicinal material, primer and identification method

A technology of molecular markers and Tripterygium wilfordii, which is applied in biochemical equipment and methods, microbial measurement/testing, DNA/RNA fragments, etc., can solve the problems of strong subjectivity and poor repeatability, and achieve high sensitivity, strong specificity, highly reproducible results

Active Publication Date: 2019-09-17
CHINA RESOURCES SANJIU MEDICAL & PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Therefore, the first technical problem to be solved by the present invention is to overcome the defects of strong subjectivity and poor repeatability of the traditional method for identifying Tripterygium wilfordii in the prior art, so as to provide a method that does not need to rely on subjective judgment, has strong specificity and high repeatability. Molecular markers, primer pairs, PCR reagents and kits for the identification of Tripterygium wilfordii
[0008] Another technical problem to be solved by the present invention is to overcome the defects of strong subjectivity and poor repeatability of the traditional identification method of Tripterygium wilfordii in the prior art, thereby providing a method that does not need to rely on subjective judgment, has a stable method, strong repeatability, and high efficiency. , Reliable method for identification of tripterygium wilfordii

Method used

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  • Molecular marker for identifying common threewingnut root medicinal material, primer and identification method
  • Molecular marker for identifying common threewingnut root medicinal material, primer and identification method
  • Molecular marker for identifying common threewingnut root medicinal material, primer and identification method

Examples

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Effect test

Embodiment 1

[0056] This example provides molecular markers and primer pairs for identifying the authenticity of Tripterygium wilfordii.

[0057] Download the nucleotide sequences of Tripterygium wilfordii and its common counterfeit products from the NCBI database. See Table 2 for the names and GenBank accession numbers. Using BioEdit software, for the gene sequences of all downloaded Tripterygium wilfordii and its counterfeit products, the comparison analysis of psbA-trnH, rbcL, ITS and matK sequences was carried out. There are no stable SNP variation sites between species and matK sequences, and there are SNP variation sites within species. Therefore, the above three sequences cannot be used for inter-species identification, and there are a large number of repetitive sequences in the psbA-trnH gene sequence itself. Suitable for PCR primer design. However, through the analysis, it was found that there were many variation sites in the ITS sequence of the genuine Tripterygium wilfordii and...

Embodiment 2

[0065]This example provides a PCR reagent for identification of tripterygium wilfordii, including a pair of primers LGTITS-715A (primer 1) and LGTITS-263A (primer 2), DNA polymerase and dNTPs.

[0066] Further, the PCR reagents include:

[0067] 2×Taqplus Master Mix II, 25μL;

[0068] Primer 1, the concentration is 6.4ng / μL, 2μL;

[0069] Primer 2, the concentration is 6.4ng / μL, 2μL;

[0070] DNA template, 200-800ng;

[0071] Add sterilized double distilled water to 50 μL.

Embodiment 3

[0073] This embodiment provides a kit for identification of tripterygium wilfordii, including the PCR reagent in embodiment 2.

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Abstract

The invention provides a molecular marker for identifying a common threewingnut root medicinal material, a primer pair, a PCR reagent, a kit and application of the molecular marker in identification of the common threewingnut root medicinal material. The molecular marker is in a nucleotide sequence shown in SEQ ID NO.3, and the primer pair can amplify the nucleotide sequence shown in SEQ ID NO.3. The invention further provides an identification method of the common threewingnut root medicinal material. Through the molecular marker, the primer pair, the PCR reagent, the PCR kit and the identification method, whether or not the molecular marker is amplified is detected so that the common threewingnut root medicinal material can be identified, the repeatability is high, and the defects of high subjectivity and poor repeatability of a traditional identification method are overcome.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a molecular marker, a primer pair and an identification method for identifying tripterygium wilfordii medicinal materials. Background technique [0002] Tripterygium wilfordii Hook.f. is the peeled and dried root of Tripterygium wilfordii Hook.f. in the family Celastraceae. one of the immunosuppressants. In modern pharmacological research, Tripterygium wilfordii also has the potential to treat Alzheimer's disease, anti-HIV, and anti-cancer effects. Because Tripterygium wilfordii has significant biological activity and contains huge medicinal value, it has become a research hotspot of scholars at home and abroad in recent years. Tripterygium wilfordii is a rhizome medicinal material. Currently, commonly used counterfeit products on the market include Dongbei Tripterygium wilfordii, kiwi fruit root, and southern snake vine. [0003] At present, there is some controversy over the pla...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12Q1/686C12N15/11
CPCC12Q1/6895C12Q1/686C12Q2565/125Y02A50/30
Inventor 杨晶凡张辉马俊谭沛王勇陈随清
Owner CHINA RESOURCES SANJIU MEDICAL & PHARMA
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