Pork component identifying molecular marker and application thereof

A molecular marker, pork technology, applied in the field of molecular biology, can solve problems such as being unsuitable for quantitative detection, and achieve the effects of improving trust, simple operation, and low time cost

Active Publication Date: 2019-10-25
SOUTH CENTRAL UNIVERSITY FOR NATIONALITIES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the different number of mitochondria in different cells, the mitochondrial DNA copy number will also vary in different tissues, so it is not suitable for quantitative detection

Method used

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  • Pork component identifying molecular marker and application thereof
  • Pork component identifying molecular marker and application thereof
  • Pork component identifying molecular marker and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1D

[0042] Example 1DNA Extraction and Amplification Experiment

[0043] 1. Experimental materials

[0044] 1.1 Animal material

[0045] Six different breeds of pigs (Sus scrofa) were used for the experiment (Enshi black pig (male), Enshi black pig (female), Enshi native pig, Huangpi native pig, Shuanghui pork, COFCO pork). The other 13 different animal materials are: cattle (Bovine), sheep (Caprinae), donkeys (Equus asinus), camels (Camelus bactrianus), chickens (Gallus domesticus), ducks (Anatinae), dogs (Canis lupus familiaris), Rat (Muroidea), Fish (Piscium), Squid (Todarodes pacificus), Shrimp (Fenneropenaeus chinensis), Scallop (Patinopectenyessoensis), Goose (Anser cygnoides orientalis).

[0046] 1.2 Enzymes and Reagents

[0047] Hieff qPCR SYBR Green Master Mix (real-time fluorescent quantitative PCR amplification premix solution) was purchased from Shanghai Yisheng Biotechnology Co., Ltd., the catalog number is 11201ES03.

[0048] Chloroform, isoamyl alcohol, ethanol,...

Embodiment 2

[0084] Example 2 Primer Screening for Specific Amplification of Pig-derived Components

[0085] 1. Experimental materials

[0086] 1.1 Pig genomic DNA

[0087] 1.2 Enzymes and Reagents

[0088] Molecular biology reagents, TAKARA Taq DNA polymerase, 10×PCR buffer, dNTP Mixture (10mM), MgCl 2 purchased from Dalian Bao Biological Company. PCR primers were synthesized by Wuhan Qingke Innovation Biotechnology Co., Ltd.

[0089] 1.3 Experimental Instruments

[0090] PCR instrument: DNA Engine Dyad Peltier Thermal Cycler (Bio-Rad, USA);

[0091] Nucleic acid electrophoresis instrument: DYYIII type YY0115-94 (Beijing Liuyi Instrument Factory);

[0092] Gel imaging system: Gel Doc XR System (Bio-Rad, USA);

[0093] Other instruments include: constant temperature water bath, electronic balance, centrifuge, vortex instrument, pure water meter, constant temperature incubator, etc.

[0094] 2. Experimental method

[0095] 2.1 Design of primers

[0096] Using the pig-specific gene (...

Embodiment 3

[0109] Intraspecific and Interspecific Analysis of Primers IPCEF370-IPCER444 for Specific Amplification of Pig-derived Components

[0110] 1. Experimental materials

[0111] 6 different breeds of pigs (Sus scrofa) (Enshi black pig (male), Enshi black pig (female), Enshi native pig, Huangpi native pig, Shuanghui pork, COFCO pork). The other 18 different animal and plant materials are: cattle (Bovine), sheep (Caprinae), donkeys (Equus asinus), camels (Camelus bactrianus), chickens (Gallus domesticus), ducks (Anatinae), dogs (Canis lupus familiaris) , Rat (Muroidea), Fish (Piscium), Squid (Todarodes pacificus), Shrimp (Fenneropenaeus chinensis), Scallop (Patinopectenyessoensis), Goose (Anser cygnoides orientalis), Soybean (Glycine max), Sesame (Sesamumindicum), Corn (Zea mays ), Arabidopsis thaliana, Nicotianatabacum.

[0112] 2. Experimental method

[0113] 2.1. Extraction of plant genomic DNA

[0114] Plant genomic DNA was extracted using QIANGEN’s small DNA extraction kit ...

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Abstract

The invention discloses a pork component identifying molecular marker and application thereof. According to a SYBR Green I dye based real-time fluorescence quantitative PCR technology, a high-specificity and high-sensitivity pork component detection method is created by taking a pig single-copy gene in a cell nucleus as the molecular marker. By adoption of the method, pigs can be accurately distinguished from other 13 animals and 5 plants, and by practical detection of 6 meat products on the market, whether porcine-derived components are contained or not can be accurately judged. The porcine specific real-time fluorescence quantitative PCR detection method provides technical references for qualitative and quantitative detection of porcine-derived components in mixed meat products.

Description

technical field [0001] The invention belongs to the field of molecular biology, in particular to a molecular marker for identifying pork components in food and its application. Background technique [0002] Pigs (Sus scrofa) are one of the most widespread mammalian species worldwide. Belonging to Animal Kingdom, Chordate Phylum, Mammalia, Artiodactyla, Suidae, Porcine genus, it is closer to primates than Rodentia. According to different geographical regions, the pig species can be classified into more than 16 different subspecies. As a subspecies of pigs, domestic pigs are the main source of pork and an important economic animal. For example, in Jianli, Hubei Province Jianli pigs from Dangyang City, Hubei Province, Qingping pigs from Dangyang City, Hubei Province, black pigs from Enshi City, Hubei Province, etc. However, most of the domestic pigs sold in the domestic market now use three imported pigs, namely Duroc, Landrace and Large White, to produce three-way hybrid com...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888
Inventor 张丽斯能武覃瑞杨怡臻严钰澳刘婷刘彦
Owner SOUTH CENTRAL UNIVERSITY FOR NATIONALITIES
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