Application of Guizhou black goat lambing major genes, primer pairs and kit

A technology of main effect gene and primer pair, applied in the field of molecular genetics and breeding, can solve the problem of reducing the activity of BMP15 promoter, and achieve the effect of improving the ability of seed production and seed supply and economic benefits

Pending Publication Date: 2021-08-20
贵州省种畜禽种质测定中心
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Studies by Chantepie et al. have shown that the BMP15 variant allele FecX can control the ovulation rate and reproductive ability of sheep, and ewes carrying homozygous FecX have strong reproductive ability and reduce the activity of the BMP15 promoter

Method used

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  • Application of Guizhou black goat lambing major genes, primer pairs and kit
  • Application of Guizhou black goat lambing major genes, primer pairs and kit
  • Application of Guizhou black goat lambing major genes, primer pairs and kit

Examples

Experimental program
Comparison scheme
Effect test

experiment example 1

[0028] Experimental Example 1 Extraction of total RNA and establishment of sequencing library

[0029] According to Trizol reagent and RNeasy RNA purification kit, total RNA was extracted from Guizhou black goat uterus, fallopian tube, and ovary tissue, and Agilent 2100 Bioanalyzer was used to detect the concentration, RIN value (RNA integrity value), 28S / 18S, and fragment size of total RNA. The mRNA with polyA structure is enriched by Oligo(dT) magnetic beads, and the RNA is broken into short fragments by ion fragmentation. The short fragment RNA is used as a template and synthesized with 6-base random primers and reverse transcriptase. cDNA first-strand, and add buffer, dNTPs, RNase and DNA Polymerase to synthesize second-strand cDNA. The end of the double-stranded DNA is filled and phosphorylated at the 5' end, and a sticky end with an "A" protruding from the 3' end is formed, and a bubbling joint with a protruding "T" at the 3' end is formed, and the ligated product The s...

experiment example 2I

[0030] Experimental example 2 Illumina sequencing

[0031] 1. Illumina sequencing process

[0032] The library was sequenced using the Illumina HiSeq sequencing platform, which was sequenced by Shenzhen Huada Genomics Co., Ltd., and the original sequence (Raw Reads) obtained was processed by removing adapter-contaminated Reads (the number of adapter-contaminated bases in the Reads was greater than 5 bp. For double end sequencing, if one end is contaminated by adapters, remove the reads at both ends); remove low-quality reads (reads whose quality value is less than 10 bases account for more than 20% of the total base number of the reads are low-quality reads) ; Remove Reads with a ratio of N greater than 5% (for paired-end sequencing, if the ratio of N at one end is greater than 5%, Reads at both ends will be removed).

[0033] 2. Sequencing results

[0034] The sequencing results showed that there were 18 samples of uterus, fallopian tubes, and ovaries in the multi-lamb grou...

experiment example 3

[0041] Experimental example 3 genome comparison and expression analysis

[0042] The clean reads were mapped to the goat (Capra hircus) reference genome (https: / / www.ncbi.nlm.nih.gov / assembly / GCF_001704415.1) using HISAT software to obtain transcriptome sequencing results. Then use RSEM to calculate the expression levels of genes and transcripts to obtain the FPKM value, which is the expression level of genes.

[0043] Gene expression analysis results: The gene expression analysis of all samples found that the overall trend of the expression distribution of each sample was consistent, and did not completely overlap, indicating that there were certain differences among the samples.

[0044] For the expression data of multiple samples, the Venn diagram was used to display the expression of genes between groups. The results showed that there were 29,522 genes expressed in the ovarian tissue of the single and multiple lamb groups, 28,741 genes were expressed in the fallopian tubes...

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Abstract

The invention discloses application of Guizhou black goat lambing major genes, primer pairs and a kit. The Guizhou black goat lambing major genes comprise up-regulation genes AMH, CYP19A1, FOXL2, FSHR, GATA4, INHBA, LHB, LHX9 and ZP3, and down-regulation genes CNOT1, DMRTA1, GHRHR, Hox-All, Oviductin, PCYT1B, SLC34A2, WDR19 and Wnt-7A; and the expression quantity of the up-regulation genes and the expression quantity of the down-regulation genes in the lambing Guizhou black goats are up-regulated and down-regulated respectively. Guizhou black goat lambing major genes are mastered, gene individuals with single-lambing property are gradually eliminated in generation breeding, gene individuals with lambing property are selected and reserved, and an important scientific basis is provided for building a new specialized lambing strain of a breeding group of the Guizhou black goats. The Guizhou black goat lambing major genes, the primer pairs and the kit has an important value for researching the polyfetal character of the Guizhou black goats, and has an important significance for improving the production performance, the breeding supply capacity, the economic benefit and the social benefit of the Guizhou local variety goats.

Description

technical field [0001] The invention belongs to the field of molecular genetic breeding, and in particular relates to the application of the main effect gene of multiple lambs of Guizhou black goat, a primer pair and a kit. Background technique [0002] Guizhou black goat is a unique local breed in Guizhou. It is mainly produced in the Bijie area of ​​Guizhou Province. It has the characteristics of pure black coat, excellent meat quality, large body, tender meat, small smell and strong stress resistance, but its fecundity is low. , The average lambing rate is 152%. Scientific research units have done a lot of research work to improve the reproductive performance of Guizhou black goats. [0003] Lambing trait is an important economic trait of sheep, and its genetic mechanism is an important basis for increasing lamb size and raising sheep production efficiency, and it is also one of the focuses of research in recent years. At present, researchers at home and abroad mostly a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888C12Q2600/124C12Q2600/158
Inventor 周迪龙清孟谭晓山杨蓉唐明艳姚敏李俊龚菲沈银李平陈昌雪刘虓
Owner 贵州省种畜禽种质测定中心
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