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SNP locus of chicken NRAMP1 gene and obtaining method and application thereof

An acquisition method and locus technology, applied in the field of poultry genetic engineering, to achieve the effect of low cost and improved resistance

Inactive Publication Date: 2017-03-15
JIANGSU INST OF POULTRY SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] After searching the existing domestic and foreign literature and patents, there has been no report on the correlation between NRAMP1 gene polymorphisms and J-subtype avian leukemia (J-ALV) resistance

Method used

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  • SNP locus of chicken NRAMP1 gene and obtaining method and application thereof
  • SNP locus of chicken NRAMP1 gene and obtaining method and application thereof
  • SNP locus of chicken NRAMP1 gene and obtaining method and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment

[0051] The above method was used to type the C125T-SNP locus of NRAMP1 gene in 100 negative and 100 Gushi chickens naturally infected with J-ALV.

[0052] 1. Sample

[0053] The blood of 100 negative and 100 Gushi chickens naturally infected with J-ALV was collected, and the DNA was extracted by the traditional phenol / chloroform method, and the DNA concentration was diluted to 50ng / μl.

[0054] 2. PCR amplification reaction and results

[0055] PCR amplification and detection results were carried out in accordance with the above method steps. The PCR product size of NRAMP1 gene was 268bp, which was consistent with the expected results (see figure 1 ). It should be noted that of 100 Gushi chicken blood samples that were determined to be negative for J-ALV, only 4 amplified the NRAMP1 gene, and 91 of the 100 positive naturally infected individuals amplified the NRAMP1 gene, which further proved the target gene It has resistance correlation with J-ALV (see Table 1).

[0056] 3. Restricti...

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Abstract

The invention discloses a SNP locus of a chicken NRAMP1 gene. The SNP locus is related to J-subtype avian leukosis resistance, and is nucleotide of the 125 locus located on a NRAMP1 gene segment, the nucleotide of the SNP locus is C or T, and the nucleotide sequence of the NRAMP1 gene segment is shown as SEQ ID NO:1. According to the chicken-NRAMP1-gene SNP locus related to the J-subtype avian leukosis resistance, a specific primer-amplification gene segment containing the SNP locus is designed according to the SNP locus of the 125 locus of the NRAMP1 gene segment, then enzyme digestion analysis is carried out with restriction enzyme, and the polymorphism of mononucleotide cam be simply, rapidly and accurately detected in a low-cost mode. The SNP locus of the chicken NRAMP1 gene is of great significance in the effect mechanism of researching chicken-NRAMP1-gene-resistant J-subtype avian leukosis, purposeful selection on chicken individuals is facilitated, and the resistance of J-ALV to offspring chickens is improved.

Description

Technical field [0001] The present invention belongs to the technical field of poultry genetic engineering, and more specifically, relates to a SNP locus of chicken NRAMP1 gene, its obtaining method and application. Background technique [0002] The NRAMP1 gene is also called solute carrier family 11 member 1 (Solute carrier family 11 member 1, SLC11A1). This gene is a relatively conservative gene. Melder DC and other studies have found that it is related to the disease resistance of humans, mice, and livestock and mainly affects the innate immunity of animals. Hu and others cloned and analyzed the chicken NRAMP1 gene, which is composed of 554 amino acids, 15 exons, 12 putative transmembrane regions and 2 glycosylation sites, which are mainly used as ion channels and transport Function. The NRAMP1 gene has 68% homology with chicken, mouse and human. Northern hybridization experiments show that: NRAMP1 gene is mainly expressed in reticuloendothelial tissues, such as phagocytes...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11C12N15/10
CPCC12Q1/6888C12Q1/6806C12Q2600/124C12Q2600/156
Inventor 范建华俞燕徐步单艳菊李尚民盛中伟章明贾雪波赵鹏
Owner JIANGSU INST OF POULTRY SCI
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