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CircRNA and application thereof in detecting intramuscular fat

A kind of intramuscular fat and circular technology, applied in DNA/RNA fragments, recombinant DNA technology, microbial measurement/inspection, etc., can solve unclear problems

Pending Publication Date: 2019-04-16
INST OF ANIMAL SCI OF CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, whether circRNA is involved in fat deposition and metabolism is still unclear

Method used

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  • CircRNA and application thereof in detecting intramuscular fat
  • CircRNA and application thereof in detecting intramuscular fat
  • CircRNA and application thereof in detecting intramuscular fat

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1 circRNA sequencing and differentially expressed circRNA selection

[0043] 1.1 Experimental animals:

[0044] In this experiment, large white pigs and Laiwu pigs with obvious differences in fat deposition were used as materials. They were raised in Laiwu Daqian Agriculture and Animal Husbandry Co., Ltd., grown and fattened under the same feeding conditions and environment, and fed according to the nutritional requirements standard (National Research Council, NRC, 1998) To feed the diet, choose 3 large white pigs (about 100kg) and 3 Laiwu pigs (about 35kg) each with a similar weight in the species, which are 180 days old, healthy and in good condition.

[0045] 1.2 Sample collection:

[0046] After slaughtering the experimental pigs, the intramuscular adipose tissue was quickly separated. To reduce RNA degradation, all processes were performed on ice. Afterwards, the tissue was cut into small pieces with sterile scissors, put into 5mL cryopreservation tubes,...

Embodiment 2

[0072] Example 2 In-depth analysis of circRNA

[0073] 2.1 Functional analysis of differentially expressed circRNAs:

[0074] After the differentially expressed circRNAs were obtained, the GO enrichment analysis was performed on the differentially expressed circRNAs using the information of the circRNA source genes. The number of differential circRNAs included in each GO entry was counted, and the significance of differential circRNA enrichment in each GO entry was calculated using the hypergeometric distribution test method. KEGG is an important public database related to Pathway. Using the KEGG database to perform Pathway analysis on differential circRNA-derived genes, you can find Pathway entries that enrich differential circRNA-derived genes, and look for potential differential circRNAs in different samples that may be related to changes in cellular pathways. . There are significant differences in intramuscular fat deposition between Large White pigs and Laiwu pigs. Thro...

Embodiment 3

[0079] Embodiment 3 real-time fluorescent quantitative PCR verification

[0080] Select circRNA_23437, 3 miRNAs (ssc-miR-370, ssc-miR-503, ssc-miR-769-3p) in the target gene of circRNA_23437, and set 20 biological replicates for each (Large White pig and Laiwu pig each 20 Example), using glyceraldehyde-3-phosphate dehydrogenase (glyceraldehyde-3-phosphate dehydrogenase, GAPDH) gene and 5SrRNA as internal references, respectively, qRT-PCR method was used to verify the expression level of the gene.

[0081] 3.1 Primer design:

[0082] circRNA_23437 amplification:

[0083] Forward primer: 5'-CACAGACTGACCGAGTGAAC-3' (SEQ ID NO.2)

[0084] Reverse primer: 5'-CCAAGTAGCAGCCAAACATG-3' (SEQ ID NO.3)

[0085] ssc-miR-370:

[0086] Specific primer: 5'-GCCTGCTGGGGTGGAACCTG-3' (SEQ ID NO.4)

[0087] ssc-miR-503 amplification:

[0088] Specific primer: 5'-TAGCAGCGGGAACAGTACTGCAG-3' (SEQ ID NO.5)

[0089] ssc-miR-769-3p amplification:

[0090] Specific primer: 5'-CTGGGATCTCTGGGGTCTTG...

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Abstract

The invention relates to a circRNA and application thereof in detecting intramuscular fat. A circRNA-seq technology and a bioinformatics method are utilized for identifying and analyzing circRNA in the intramuscular fat of Laiwu pigs and large white pigs, circRNA_23437 relevant to regulation and control of pig fat decomposition is found out, the mechanism of the circRNA in regulating fat decomposition and the fat metabolism function is further known, and a theoretical basis is laid for improving the quality of pork.

Description

technical field [0001] The present invention relates to the field of biotechnology, in particular to a porcine circular RNA and its application in detecting intramuscular fat, and more specifically to circRNA_23437 and its application. Background technique [0002] Pork is the main source of people's dietary structure. According to the forecasts of USDA economists and OECD-FAO, compared with 2014, global pork production will increase by at least 10.5% by 2023. With the increase of people's demand, the requirements for pork meat quality are getting higher and higher, mainly including meat color, tenderness, flavor and juiciness. Intramuscular fat is an important factor affecting meat quality. Over the years, during the breeding process aimed at high lean meat percentage and rapid growth, the quality of pork has declined. The main reason is the reduction of fat, especially the reduction of intramuscular fat deposition. However, excessive fat deposition will lead to obesity an...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12N15/113
CPCC12Q1/6888C12Q2600/124C12Q2600/158C12Q2600/178
Inventor 苗向阳李嫒解领丽黄万龙张秀秀
Owner INST OF ANIMAL SCI OF CHINESE ACAD OF AGRI SCI
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