Intramuscular fat related lncRNA (long non-coding Ribonucleic Acid) and application thereof

A kind of intramuscular fat, a pair of technology, applied in the field of molecular biology, can solve the problems of lower intramuscular fat content and lower pork quality

Active Publication Date: 2018-06-01
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

[0002] Pork is the main meat product in my country's dietary structure. Adipose tissue has an important impact on meat quality, especially intramuscular fat, including meat appearance, flavor, water holding capacity, tenderness, etc. However, in the pursuit of high lean meat percentage During the long-term selection process of pig carcasses, the intramuscular fat content decreased and the pork quality decreased

Method used

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  • Intramuscular fat related lncRNA (long non-coding Ribonucleic Acid) and application thereof
  • Intramuscular fat related lncRNA (long non-coding Ribonucleic Acid) and application thereof
  • Intramuscular fat related lncRNA (long non-coding Ribonucleic Acid) and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1 Sample collection preparation and experimental design

[0037] 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

[0038] Example 2 Extraction and quality control of sample total RNA

[0039] 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

[0040] Example 3 cDNA library construction and RNA sequencing

[0041] strand-specific cDNA library

[0042] (1) Ribo-zero kit removes rRNA

[0043] (2) RNA fragmentation

[0044] (3) Double-stranded cDNA synthesis and purification

[0045] (4) End repair, adding A base

[0046] (5) Sequencing adapter connection

[0047] (6) Enrichment and purification of DNA fragments

[0048] (7) Library Quality Inspection

[0049] (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).

[0050] RNA-Seq (Illumina Sequence)

[0051] 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 intramuscular fat related lncRNA (long non-coding Ribonucleic Acid) and application thereof. The invention finds out XLOC_004398 related to intramuscular fat of pork; a targetgene NAP1L3 for differentially expressing the lncRNA is analyzed and predicated through co-expression network analysis and a trans regulation effect and is verified. The invention provides certain evidences for breeding high-meat-quality livestock and poultry varieties and treating and preventing fat metabolism related diseases, and a new target is explored.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and in particular relates to an lncRNA related to pig intramuscular fat and an application thereof. Background technique [0002] Pork is the main meat product in my country's dietary structure. Adipose tissue has an important impact on meat quality, especially intramuscular fat, including meat appearance, flavor, water holding capacity, tenderness, etc. However, in the pursuit of high lean meat percentage During the long-term breeding process, the intramuscular fat content in pig carcasses decreased, and the pork quality decreased. As an excellent local pig breed resource in my country, Laiwu pig has valuable germplasm characteristics such as high fecundity and excellent meat quality. It is a typical representative of local black pigs in Shandong Province. Its carcass fat content is relatively high, and its muscles have bright red flesh color. Water properties, more importantly, rich in...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12Q1/6841C12N15/113
CPCC12Q1/6841C12Q1/6888C12Q2600/124C12Q2600/178
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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