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Method for identifying thelenota ananas by means of special peptide fragment group

A kind of plum ginseng, specific technology, applied in the biological field, can solve the problems of lack of technical level, deceiving consumers and so on

Active Publication Date: 2015-12-02
OCEAN UNIV OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Due to the lack of a particularly effective method of identifying sea cucumber species at present, the method of identifying sea cucumber species is only based on traditional appearance observation, lacking technical support, resulting in many unscrupulous merchants cheating consumers by shoddy goods and false ones. Therefore, it is necessary to An efficient and accurate method for identifying sea cucumbers

Method used

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  • Method for identifying thelenota ananas by means of special peptide fragment group
  • Method for identifying thelenota ananas by means of special peptide fragment group
  • Method for identifying thelenota ananas by means of special peptide fragment group

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1. Sequence source and alignment information of specific polypeptides

[0037] 1. The source of the unique peptide ADIAEDSLK of plum ginseng

[0038] The unique polypeptide ADIAEDSLK of plum ginseng is derived from the protein tropomyosin [Apostichopusjaponicus]. Its Accesionnumber on NCBI is gi|302340969. The sequence is shown in SEQIDNO.5 below. Analysis shows that Glu at position 6 of ADIAEESLK has been mutated into Asp, and the resulting polypeptide ADIAEDSLK was compared by NCBIblast, and it was not found to be consistent with any other species:

[0039] SEQIDNO.5:

[0040] MDTIKKKLSQLKADKEKALDEKDVAEASMKEAMERVEQVNDENKELQTRIKQLETELDDTSEKLNTTVIKCEAAEKAQQTAEEEMANLQRKLQLTEEELSRSEERVADLQSKYTDIEQSSEENERQRKVLESRSAADDERMSELETQVMSSKTSLEDSDRKYDEASRKLTVTEEELARSEERSAAFESSLSQMKEELHQLHNNVKSLEAQEEKFTENEEMYEKKVRDLEDKLKVAEDRADIAEESLKSLKTSLDQLEDELMIEKEKVREMTEEMERTIQELNFEV

[0041] Adopt ABCIEX 5600 mass spectrometer (AB5600) detects the fragmentation and mass spectra of the target p...

Embodiment 2

[0066] Example 2, Sea cucumber sample processing and detection steps

[0067] The steps to analyze the sea cucumber sample to be tested include

[0068] (1) Sample pre-processing steps:

[0069] (1) Weigh 1g of sea cucumber sample and homogenize it into powder state, add 10mL protein extract (8M urea, 50mMNH 4 HCO 3 ) Extract protein by shaking, centrifuge at high speed and low temperature (20000r / min), take 200ul supernatant and transfer to 1ml EP tube;

[0070] (2) Take 2μL of 1mol / LDTT (0.154g dithiothreitol DTT dissolved in 1mL of deionized water) and add it to 100μL of the above protein solution to react at 37°C for 1 hour;

[0071] (3) Take 10 μL of freshly prepared IAA (accurately weigh 0.185 g of iodoacetamide IAA and dissolve in 1 mL of deionized water, and configure it into 1 mol / L IAA mother liquor for use. The configuration process needs to be protected from light. In the above reaction solution that has been cooled to room temperature, react at room temperature and avoid l...

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Abstract

The invention relates to a method for identifying thelenota ananas by means of a special peptide fragment group, and provides a group of contrast polypeptide used for identifying the thelenota ananas through mass-spectrography. The sequences of the polypeptide are shown as SEQ ID NO.1:ADIAEDSLK, SEQ ID NO.2:AGPAGDDAPR, SEQ ID NO.3:MPIPLLEGK and SEQ ID NO.4:LDIPLLEGK. Through identification, the peptide fragment of which the sequence is shown as the SEQ ID NO.1 ADIAEDSLK is the special peptide fragment of the thelenota ananas and is not expressed in other species; the SEQ ID NO.2:AGPAGDDAPR, SEQ ID NO.3:MPIPLLEGK and SEQ ID NO.4:LDIPLLEGK exist in the other species, but are not expressed in stichopus japonicus, acaudina molpadioidea, isostichopus badionotus and isostichopus fuscus; the peptide fragments are the special peptide fragments of the thelenota ananas.

Description

Technical field [0001] The present invention belongs to the field of biotechnology, and specifically relates to a method for identifying plum ginseng (Thelenotaananas) by using a specific peptide group. Background technique [0002] Sea cucumbers are rich in acid mucopolysaccharides, which have immunomodulators, anticoagulant, blood sugar control and blood lipid lowering effects on the human body. Therefore, sea cucumber enjoys the reputation of "nutrition treasure house", and modern people regard sea cucumber as the "first of the eight treasures of seafood". There are many kinds of Shanghai ginseng in the market. Different sea cucumbers vary greatly in price due to their nutritional value and taste. Plum ginseng (Thelenotaananas) has high economic value like gray sea cucumber. It is not only a tonic, but also can cure diseases and fight cancer. It has anti-aging effect, contains high protein, rich minerals, and does not contain cholesterol. It is an ideal tonic. Traditional Ch...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/62
Inventor 李八方张鸿伟赵雪
Owner OCEAN UNIV OF CHINA
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