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A kind of molecular identification method of Uncaria housi

A technology for molecular identification and Uncaria chinensis, which is used in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., to achieve the effect of making up for limitations and accurate molecular identification.

Active Publication Date: 2019-10-15
GUANGDONG PHARMA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The technical problem to be solved by the present invention is to overcome the defects and deficiencies in the identification of Uncaria rhynchophylloides (Uncaria rhynchophylloides) by traditional methods such as morphology, histology and chemical composition analysis, and to provide an accurate method for identification of Uncaria rhynchophylloides. High and fast molecular biology methods

Method used

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  • A kind of molecular identification method of Uncaria housi
  • A kind of molecular identification method of Uncaria housi
  • A kind of molecular identification method of Uncaria housi

Examples

Experimental program
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Embodiment 1

[0036] The molecular identification of embodiment 1 Uncaria houshou

[0037] 1. Genomic DNA extraction:

[0038] (1) Samples to be tested: multiple samples identified as Uncaria rugosa according to morphology were collected from different origins. This embodiment uses some samples as examples for illustration, as shown in Table 1.

[0039] (2) Take 1 g of freshly collected leaves or freshly collected leaves that have been quickly dried on silica gel and then sealed and preserved, and quickly ground into fine powder under liquid nitrogen conditions, and use the improved CTAB method to extract the genomic DNA of the plant to be tested .

[0040] Table 1 Sampling locations of Uncaria housi samples

[0041]

[0042]

[0043] 2. Amplification of sequence fragments in the ITS region:

[0044] (1) Using primers ITS5 and ITS4 to amplify the total DNA of Uncaria rDNA ITS sequence.

[0045] The sequences of primers ITS5 and ITS4 are as follows:

[0046] Primer ITS5 (as shown ...

Embodiment 2

[0071] 1. Use wax leaf specimens of Uncaria rhynchophylloides from different origins to study and verify the variation sites that are conducive to identification.

[0072] The method is basically the same as that in Example 1, except that the wax leaf specimen of Uncaria marquis is used as a sample, the genomic DNA is extracted, the sequence of the ITS region is amplified by PCR, sequenced, compared and analyzed for identification.

[0073] 2. As a result, the above-mentioned 7 mutation sites can also be specifically detected, and through sequence comparison, it is found that the sequence of the ITS region of Uncaria housiensis satisfies at least one of the following conditions:

[0074] a. 120 bits are A and 547 bits are T;

[0075] b. 12 bits are A and 569 bits are not inserted with T;

[0076] c. 105 bits are C and 86 bits are T;

[0077] d. 105th place is C and 450th place is C.

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Abstract

The invention discloses a molecular identification method of Uncaria rhynchophylloides F. C. How. The method comprises the following steps: on the basis of ITS sequence (disclosed as SEQ ID NO.1) of Uncaria rhynchophylloides F. C. How obtained for the first time, carrying out amplification by using primers ITS5 and ITS4 by using total DNA (deoxyribonucleic acid) of the sample to be detected as a template; and carrying out comparison analysis on rDNA ITS sequence and the sequence disclosed as SEQ ID NO.1: carrying out comparison on at least 2 bases from matched 12th, 86th, 105th, 120th, 450th, 547th and 569th bases, and identifying whether the sample to be detected is Uncaria rhynchophylloides F. C. How according to the comparison result. The sites above are unique site information distinguished from other plants of wild gambier, and can be used for carrying out quick and accurate molecular identification on Uncaria rhynchophylloides F. C. How on the molecular biology level. The method provides necessity guarantees for specified drug application and safe drug application of the traditional Chinese medicine Uncaria rhynchophylloides F. C. How.

Description

technical field [0001] The invention belongs to the technical field of molecular identification. More specifically, it relates to a molecular identification method of Uncaria hunchia. Background technique [0002] Uncaria rhynchophylloides is a plant of the Rubiaceae Uncaria genus, which is mainly distributed in Guizhou, Yunnan, Guangdong, Guangxi and other regions of China. It is a commonly used medicine in traditional Chinese medicine and has a long history of medicinal use. "Compendium of Materia Medica" records: "Uncaria is also a medicine for hand and foot Jueyin. Foot Jueyin (liver meridian) governs wind, hand Jueyin (pericardium meridian) governs fire, Uncaria connects the pericardium to liver wood, and the wind is calm and the fire is extinguished." , all diseases will be cured by themselves." It has the effects of relieving wind and calming convulsions, clearing away heat and calming liver. The research on chemical constituents shows that the main effective const...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12N15/11
Inventor 朱爽王业胜曾常青周林李奇威
Owner GUANGDONG PHARMA UNIV
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