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A new lncRNA extracted from porcine intramuscular adipose tissue and its application

A technology of intramuscular fat and tissue, applied in the field of molecular biology, can solve the problems of lncRNA expression profile and function, which are rarely studied

Active Publication Date: 2019-11-05
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

As a regulatory non-coding RNA, it plays an important regulatory role in pig fat metabolism and adipogenic differentiation. At the same time, there are more and more studies on lncRNA regulation of fat metabolism-related diseases. However, different breeds of pigs with significant differences in fat deposition The expression profile and function of lncRNA in intramuscular adipose tissue are rarely studied

Method used

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  • A new lncRNA extracted from porcine intramuscular adipose tissue and its application
  • A new lncRNA extracted from porcine intramuscular adipose tissue and its application
  • A new lncRNA extracted from porcine intramuscular adipose tissue and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Example 1 Sample collection preparation and experimental design

[0040] After the experimental pigs were slaughtered, the fat tissue in the longissimus dorsi muscle was quickly collected, cut into small pieces, filled into 5mL cryopreservation tubes, frozen in liquid nitrogen, and then transferred to a -80°C refrigerator for long-term storage for total RNA The extraction, the experiment set up 3 groups, identified the lncRNA in the large white pig intramuscular adipose tissue (D_JN) and the Laiwu pig intramuscular adipose tissue (L_JN), and analyzed their intramuscular adipose tissue gene expression profiles, each sample set 3 repetitions.

Embodiment 2

[0041] Example 2 Extraction and quality control of sample total RNA

[0042] Take out the same amount of cryopreserved adipose tissue samples respectively, and use mirVana according to the instruction manual TM The RNA extraction kit was used to extract total RNA from each adipose tissue sample, and the isolated total RNA sample was stored in a -80°C refrigerator. Use the NanoDrop 2000 spectrophotometer to measure the concentration and OD260nm / OD280nm value of the RNA sample, and control it between 1.9 and 2.1, use the Bioanalyzer 2100 to evaluate the quality of the total RNA, and control RIN>=7 and 28S / 18S>=0.7, use RNase-free DNase I eliminates potential genomic DNA contamination.

Embodiment 3c

[0043] Example 3 cDNA library construction and RNA sequencing

[0044] strand-specific cDNA library

[0045] (1) Ribo-zero kit removes rRNA

[0046] (2) RNA fragmentation

[0047] (3) Double-stranded cDNA synthesis and purification

[0048] (4) End repair, adding A base

[0049] (5) Sequencing adapter connection

[0050] (6) Enrichment and purification of DNA fragments

[0051] (7) Library Quality Inspection

[0052] (8) A total of 6 cDNA libraries were established in this study, namely D_JN_1, D_JN_2, D_JN_3 (Large White pig intramuscular adipose tissue cDNA library) and L_JN_1, L_JN_2, L_JN_3 (Laiwu pig intramuscular adipose tissue cDNA library).

[0053] RNA-Seq (Illumina Sequence)

[0054] After passing the quality inspection of the library, the Illumina HiSeqTM 2500 sequencing platform was used to perform sequencing analysis on the cDNA library using Paired-end Sequencing, and the off-machine data was the original sequencing data raw reads.

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Abstract

The invention discloses new lncRNA extracted from pig intramuscular fat tissue and application of the new lncRNA. A RNA-seq technology and a bioinformatics method are adopted to compare the gene expression profiles of the intramuscular fat tissues of a large white pig and a Laiwu pig, and the key differentially expressed lncRNA related to adipogenic differentiation and lipid metabolism are identified and screened out. Molecular verification experiments indicate that the lncRNA and the target genes PLEKHG3, SERPINB7, SPTLC3 and TMOD2 thereof are all closely related to the pig intramuscular fat.The invention lays a theoretical basis for the pig lipidosis mechanism study and livestock fat tissue lncRNA study, and has an important application prospect in molecular breeding of livestock.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and specifically relates to a new lncRNA extracted from pig intramuscular adipose tissue and its application, and more specifically relates to the application of XLOC_007365 in breeding high-quality livestock breeds. Background technique [0002] Adipose tissue has an important impact on meat quality, especially intramuscular fat, including meat appearance, flavor, water holding capacity, tenderness, etc. The fat content in the meat is reduced, and the pork quality is reduced. At the same time, adipose tissue is a tissue that stores energy. Excessive deposition of adipose tissue in the human body can lead to obesity and imbalance of energy metabolism, which will cause obesity-related diseases, such as type 2 diabetes, insulin resistance, cardiovascular disease and certain cancers, seriously threatening human health. Therefore, human health problems caused by meat quality and obesity h...

Claims

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Application Information

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IPC IPC(8): C12N15/113C12Q1/6888G01N33/68
CPCC12N15/113C12Q1/6888C12Q2600/124C12Q2600/158C12Q2600/178G01N33/68
Inventor 苗向阳黄万龙张秀秀李嫒解领丽
Owner INST OF ANIMAL SCI OF CHINESE ACAD OF AGRI SCI
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