Molecular marker related to chicken heat stress tolerance, primer combination and detection and identification method thereof

A molecular marker and chicken heat stress technology, which is applied in the fields of genetic engineering and genetic breeding, can solve the problem that the molecular markers of chicken heat stress tolerance are still few and other problems, and achieve the effect of simple operation and low cost.

Pending Publication Date: 2022-08-09
JIANGSU INST OF POULTRY SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, there are few studies on molecular markers of heat stress tolerance in chickens at home and abroad, and only a few studies have reported potential genetic markers that can be used as heat stress resistance in chickens

Method used

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  • Molecular marker related to chicken heat stress tolerance, primer combination and detection and identification method thereof
  • Molecular marker related to chicken heat stress tolerance, primer combination and detection and identification method thereof
  • Molecular marker related to chicken heat stress tolerance, primer combination and detection and identification method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Four local breed chickens grown in different environments in China were selected, including 50 Wenchang chickens, Dongxiang green-shelled chickens, Tibetan chickens and Lindian chickens. 2 mL blood samples were collected from the fin vein in EDTA-Na anticoagulation tubes, and the phenol-chloroform method was used. Genomic DNA extraction was performed. Whole genome sequencing was performed on the Illumina Hiseq PE150 platform after DNA quality control. By omitting linker sequences (>10nt for calibration with adapters, allowing ≤10% mismatches), remove low-quality sequences including ≥10% unrecognized nucleotides or >50% with base sequencing quality <5 , to remove PCR repeats that may be generated during library construction. Using the BWA (Burrows-Wheeler Aligner) (version 0.7.8) software package, the remaining high-quality paired sequences were matched with the chicken reference genome Gallus_gallus-5.0 with the command mem-t4-k32 M; at the same time, in order to reduc...

Embodiment 2

[0030] In order to further investigate whether the allelic mutation frequency pattern of the above two SNP loci is related to the adaptation to high temperature environment, four other local chicken breeds in China, including Minqing Maojiao chicken (MQMC) and Guangxi Longshengfeng chicken (LSFC), were selected in the experiment. ), Shandong Jining Bairi Chicken (JNC) and Beijing Oil Chicken (BJYC), 100 each, as well as 191 Brazilian local chickens (Brazilian), 1439 Ghana local chickens (Ghana) and 1399 Tanzania local chickens (Tanzania). 2 mL blood samples were collected intravenously in EDTA-Na anticoagulant tubes, and genomic DNA was extracted by phenol-chloroform method. After passing the DNA quality control, the target fragments of the two SNP sites related to high temperature tolerance were amplified. The upstream and downstream primers for the amplification of the two sequences were as follows:

[0031] Primer with base at position 40827385 of chromosome 5:

[0032] Up...

Embodiment 3

[0052] In order to further explore whether the allele mutation frequencies of the above two SNP loci are suitable for the identification of heat stress tolerance populations, 180 35-week-old layer A and 180 layer B were selected in the experiment. Feed a basal ration, ad libitum. In this experiment, a cyclic heat stress test was performed, and the temperature was set as: 20:00-00:00, 00:00-06:00 was set to 28°C, 06:00-10:00, 16:00-20:00 was set to 32°C, 10:00-16:00 set to 35°C. The trial period was 2 weeks. On the 14th day of the test, the production performance was counted, and the blood was collected to extract DNA, and then amplified according to the two pairs of primer sequences and PCR conditions in Example 2, and then sent to the company for sanger sequencing to detect the genes of the two SNP sites in Example 2. type.

[0053] The results are shown in Table 3. The rectal temperature, respiratory rate, egg production rate and mortality rate of chickens in group B were...

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Abstract

The invention relates to an SNP (Single Nucleotide Polymorphism) molecular marker related to chicken heat stress tolerance. The SNP molecular marker corresponds to the 40827385th site and the 40829460th site of the fifth chromosome of a chicken reference genome (Gallus 5.0) on NCBI (National Center of Biotechnology Information). Wherein when the two loci are CC and TT, the two loci can be used as dominant genotypes to select the heat stress tolerance of the chicken, an effective, simple and easy-to-operate and low-cost molecular genetic marker is provided for chicken molecular marker-assisted breeding of heat stress tolerance groups, and an effective molecular genetic marker is provided for improvement of the heat stress tolerance of the chicken.

Description

technical field [0001] The invention relates to the fields of genetic engineering and genetic breeding, in particular to a molecular marker related to chicken heat stress tolerance, a primer combination and a detection and identification method thereof. Background technique [0002] In 2022, the national key research and development plan will include the adaptability of agricultural biological environment into the application guidelines, in response to the country's measures to ensure people's food security. Poultry products are an important part of people's food, and it is very important to ensure their quality and safety. The poultry farming system is affected by a variety of climatic factors, but due to today's public awareness and abundant scientific information intake, the quality and safety issues of poultry products related to heat stress have become particularly prominent. In the context of modern intensive poultry farming, heat stress is of particular concern in po...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888C12Q2600/156C12Q2600/124
Inventor 施寿荣邵丹童海兵胡艳王强张珊肖蕴祺沈一茹张安陈静龙
Owner JIANGSU INST OF POULTRY SCI
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