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Application of reagent for detecting expression quantity of MSTRG.111777 gene in judgment of ovulation condition of sheep and reagent

A technology of MSTRG.111777, gene expression amount, applied in the field of genetic engineering

Pending Publication Date: 2022-05-31
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, both FSH and LH receptors are G protein-coupled receptors, and they exhibit a cascade amplification effect in the signal transduction process in cells, but there is no literature explaining the role of lncRNA in FSH and LH signal transduction in detail

Method used

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  • Application of reagent for detecting expression quantity of MSTRG.111777 gene in judgment of ovulation condition of sheep and reagent
  • Application of reagent for detecting expression quantity of MSTRG.111777 gene in judgment of ovulation condition of sheep and reagent
  • Application of reagent for detecting expression quantity of MSTRG.111777 gene in judgment of ovulation condition of sheep and reagent

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Test group

[0042] In the core production area of ​​small-tailed Han sheep in Yuncheng, Shandong Province, 12 multiparous small-tailed Han sheep with similar body size and body condition were selected, among which 6 small-tailed Han sheep with three consecutive litters with more than 2 lambs were selected as In the multi-lamb group, 6 small-tailed Han sheep with less than 2 lambs in three consecutive litters were used as the single-lamb group, see Table 1 for details. These ewes are about 3 years old and weigh about 70 kg. In the spring, all selected test ewes were placed in the vagina for 12 days using CIDR (300 mg progesterone) to perform estrus synchronization. In the absence of ingestion of exogenous sex hormones, 5 mL of vitamin AD was injected intramuscularly to protect the vaginal epithelial mucosa. Free access to food and water was guaranteed during the experiment. The pituitary glands were collected from 3 single-lamb and multiple-lamb Small-tailed Han shee...

Embodiment 2

[0047] RNA extraction, library construction and sequencing

[0048] The total RNA of the pituitary was extracted using TRIzol reagent (Thermo Fisher Scientific, Waltham, MA, United States), and the extracted Spectrophotometer (IMPLEN, Westlake Village, CA, USA) and RNA analysis kit (Thermo Fisher Scientific) was used to detect the purity and concentration of total RNA, and Agilent 2100 RNA Nano 6000 Assay Kit was used to determine the RIN value of RNA. The OD260 / 280 ratio of each sample was between 1.8 and 2.1, and the RIN≥7, indicating that the RNA integrity and quality were good, which met the requirements for sequencing on the machine. The test results are shown in Table 2.

[0049] Table 2 Total RNA purity, concentration, integrity

[0050]

[0051]

[0052] For each pituitary sample, 3 μg of total RNA was used as the initial amount to construct mRNA and lncRNA libraries, and rRNA in the total RNA was removed using Ribo-ZeroTM GoldKits (Epicentre, USA). The Inde...

Embodiment 3

[0054] Data Analysis and qRT-PCR Validation

[0055] 1. Data Quality Control and Assembly

[0056] Use bcl2fastq (v.2.17.1.14) to process Illumina high-throughput sequencing files, and convert raw image files into raw sequencing data based on base calling technology, namely rawreads. A Perl script is used to filter rawreads for high quality reads. The steps include: 1) Reads with adapter contamination, mainly refer to reads with more than 5 bp of contaminated bases. 2) Low-quality reads (Phred Quality Score <5%). 3) Reads containing more than 5% of unknown bases. 4) Reads matched to rRNA. Phred Quality Score represents different base sequencing error rates, for example, Q20 and Q30 represent base sequencing error rates of 1% and 0.1%, respectively. The filtered clean reads were aligned to the sheep reference genome (Oarv.4.0) using HiSAT2 (v2.0.5), and the alignment results are shown in Table 3. The aligned reads were assembled using StringTie (v1.3.2d). The parameters ...

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Abstract

The invention relates to application of a reagent for detecting MSTRG.111777 gene expression quantity in judging ovulation condition of sheep and the reagent, and belongs to the technical field of gene engineering. The invention provides application of a reagent for detecting the expression quantity of the MSTRG.111777 gene in judging ovulation conditions of sheep. By means of the application, the ovulation condition of the sheep can be judged, and the multi-lamb character and the single-lamb character of the sheep can be distinguished.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to the application of a reagent for detecting the expression level of the MSTRG.111777 gene in judging the ovulation condition of sheep and the reagent. Background technique [0002] At present, my country's sheep breeding is intensive house feeding and semi-house feeding. Although the number of large-scale breeding is increasing year by year, the development speed is slow. Due to the extremely high feeding cost and lack of standardization, mutton cannot be of high quality and price. It is the most important reproductive trait. Therefore, it has become an urgent need for the development of mutton sheep in my country, and it is also the main direction of breeding. The litter size trait of sheep is a low heritability trait, and the sheep generation interval is long, so it is difficult to improve the efficiency of the selection reaction by ordinary phenotypic selection. The...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888
CPCC12Q1/6888C12Q2600/158C12Q2600/124C12Q2600/178Y02P60/87
Inventor 储明星陈思王翔宇郭晓飞赵福林狄冉贺小云
Owner INST OF ANIMAL SCI OF CHINESE ACAD OF AGRI SCI
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