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Tenebrio molitor L. YM47 protein, coding gene of protein and application

A technology of Tenebrio molitor and protein, applied in application, genetic engineering, plant genetic improvement, etc., can solve problems such as increasing the difficulty of positioning and screening insect-specific allergens, having no related species of Tenebrio molitor, and interfering with allergy prediction and diagnosis. , to achieve the effect of reducing the risk of development and utilization

Pending Publication Date: 2019-10-18
SICHUAN AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] In the prior art, there are very few studies on insect allergenic proteins, and there is no related research on related species of Tenebrio molitor as a reference for comparative research
In addition, when a certain insect food is consumed daily, the prediction and diagnosis of allergies will be interfered by the cross-effect of other insect-derived proteins or foods containing allergens
At the same time, most of the existing studies have failed to clarify whether there is an interaction between inhalant allergy, contact allergy, and ingestion allergy, or whether there is a possibility of various comorbid diseases in such individuals with allergic symptoms
The above situation increases the difficulty of the location screening of insect-specific allergens, and leads to the fact that most of the insect allergens measured in the existing related studies are mostly detected in the case of cross allergies

Method used

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  • Tenebrio molitor L. YM47 protein, coding gene of protein and application
  • Tenebrio molitor L. YM47 protein, coding gene of protein and application
  • Tenebrio molitor L. YM47 protein, coding gene of protein and application

Examples

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Embodiment 1

[0023] This embodiment discloses the extraction and purification of Tenebrio molitor total protein, specifically as follows:

[0024] A. Tenebrio molitor total protein extraction

[0025] Live and mature mature larvae of Tenebrio molitor were selected, freeze-dried in liquid nitrogen and ground into powder. Then, 150 mL of lysis buffer (150 mL of phosphate buffered saline solution containing 0.5% TritonX-100 at pH 7.4 and 3 EDTA-free protease inhibitor tablets produced by Thermo Fisher Scientific) was added to 10 g of insect powder, and the MSE Soniprep150 ultrasonic homogenizer ultrasonic breaker. When using an ultrasonic homogenizer, divide the mixture of insect powder and 150mL lysis buffer into four 50mL centrifuge tubes, and place the centrifuge tubes in a box with ice to keep the temperature low during the entire ultrasonication process. At the same time, pause the instrument to cool for 30 seconds after every 15 seconds of treatment, and cycle like this until the inse...

Embodiment 2

[0032] This example discloses SDS-PAGE detection of protein samples.

[0033] Referring to the experimental method of the experimental operation manual provided by Thermo Company and optimized and improved, the total protein sample of Tenebrio molitor, the protein component samples (4 samples) obtained by the first separation and purification and the higher concentration obtained after the second separation and purification were selected. The protein component samples (9 samples), a total of 14 samples. The 14 samples are: total protein, peak1, peak1Q5, peak1Q6, peak2Q7, peak2Q8, peak2Q9, peak3, peak3Q4, peak3Q7, peak4, peak4Q3, peak4Q4, peak5.

[0034] Take 6.5 μL of each sample and add 3 μL sample buffer (Pierce TM SDS-PAGE Sample Prep Kit) and 0.5 μL of dithiothreitol (DTT), the mixture was placed in a PCR instrument and heated at 70°C for 12min. At the same time, prepare 1L electrophoresis buffer (50mL NuPAGE SDS solution and 950mL ultrapure water), wash the gel (NuPAGE...

Embodiment 3

[0041] The present embodiment discloses the extraction method of human serum for test, specifically:

[0042] According to the proposed biomedical research plan, 5 subjects were screened out each with history of allergy to Tenebrio molitor (experimental group) and without history of allergy to Tenebrio molitor (positive control group). The specific method is: 60 volunteers with experience of eating Tenebrio molitor were screened out on a voluntary basis, and asked whether they had any adverse reactions related to allergies after eating Tenebrio molitor, and they were in good health, without respiratory diseases, blood system Diseases and other medical history that do not meet the screening conditions of the test, and the volunteers are not smokers or alcoholics. At the same time, do not participate in any other clinical trials within four weeks of the start of this trial.

[0043] Use a 10mL vacuum glass blood vessel (red cap) to collect 40mL of whole blood (4 tubes, 10mL / tub...

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Abstract

The invention discloses a tenebrio molitor L. YM47 protein, a gene for coding the protein and application of the tenebrio molitor L. YM47 protein and the coding gene of the protein. The YM47 protein is a tenebrio molitor L. sensitization protein which can cause the human allergy. The amino acid sequence of the tenebrio molitor L. YM47 protein is shown in SEQ ID NO:3. The nucleotide sequence of thegene for coding the YM47 protein is shown in SEQ ID NO:4. The invention discloses the tenebrio molitor L. specificity sensitization protein and the coding gene thereof for the first time. Breeding ofsensitization-free tenebrio molitor L. can be achieved. Through the amino acid sequence of the sensitization protein and a gene sequence corresponding to the amino acid sequence, the sensitization protein of other resource insects can be speculated, and the risks of development and utilization of insect source protein food caused by potential sensitization of insect protein can be reduced to a certain degree.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to Tenebrio molitor YM47 protein, its coding gene and application. Background technique [0002] Tenebrio molitor L. (Tenebrio molitor L.) of the genus Coleoptera, Pedidae, commonly known as mealworms, is the third largest economic insect after silkworms and bees. It has the advantages of high protein content in the body, rich variety, good nutritional and health effects, etc., and is considered to be a new protein source that has not been fully developed and utilized. However, such protein foods, like other protein-containing foods, have the potential risk of causing allergies in a small number of eaters (Sicherer, 2011). How to predict, diagnose and solve this limitation problem has become an urgent need to alleviate the shortage of protein resources. Existing studies on four orders and ten common edible insects have found that tropomyosin and arginine kinase (tropomyosin...

Claims

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Application Information

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IPC IPC(8): C07K14/435C12N15/12
CPCC07K14/43563
Inventor 王南溪杨伟杨桦杨春平方睿钱丽芝朱晓庆管凤荣王保新
Owner SICHUAN AGRI UNIV
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