Excellent slaughter trait molecular genetic marker of broiler chicken and application of excellent slaughter trait molecular genetic marker

A molecular genetic marker and trait technology, applied in the application field of broiler chicken breeds

Inactive Publication Date: 2015-05-06
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, after searching, there is no report on the application of plin1 gene SNPs as genetic markers, especially as molecular genetic markers for good slaughtering traits of broiler chickens to screen or predict broiler chicken breeds with good slaughtering traits

Method used

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  • Excellent slaughter trait molecular genetic marker of broiler chicken and application of excellent slaughter trait molecular genetic marker
  • Excellent slaughter trait molecular genetic marker of broiler chicken and application of excellent slaughter trait molecular genetic marker
  • Excellent slaughter trait molecular genetic marker of broiler chicken and application of excellent slaughter trait molecular genetic marker

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Embodiment 1: Cloning of partial DNA sequences of chicken plin1 gene of different breeds

[0026] 1. Collection and processing of chicken blood: Randomly select disease-free healthy chickens, including 1478 Luqin No. 1 chickens, 616 Luqin No. 3 chickens, and 68 Shiqi chickens. Blood is collected from the vein under the wing, and EDTA is used for anticoagulation deal with.

[0027]2. Genomic DNA extraction: DNA was extracted using Tiangen blood DNA extraction kit. Use 200 μL of fresh, frozen or blood with various anticoagulants, add 20 μL proteinase K solution, mix well; then add 200 μL buffer GB, mix well and place at 70°C for 10 minutes; add 200 μL absolute ethanol, shake and mix well At this time, flocculent precipitates will appear; transfer all the obtained solution and flocculent precipitates to the adsorption column CB3 in the collection tube, centrifuge at 12000rpm for 30 seconds and discard the waste liquid; add 500μL buffer GD, centrifuge at 12000rpm for 30 se...

Embodiment 2

[0037] Embodiment 2: Use NspI, BstNSI and NspHI enzymes to carry out specific digestion of the PCR product in embodiment 1

[0038] 1. Use NspI, BstNSI and NspHI enzymes to specifically digest the target gene. The action site of this type of specific enzyme is: 5'...RCATG ↓ Y…3’ or 3’…Y ↑ GTACR...5'.

[0039] 2. See Table 3 for the enzyme digestion reaction system.

[0040] Table 3 NspI enzyme digestion reaction system

[0041]

[0042] After mixing the solutions of the reaction system according to the above system, place them in a constant temperature water bath at 37°C to react for 3-5 hours, so that the enzyme cleavage reaction can fully proceed, and then carry out subsequent tests.

[0043] 3. The digested products were analyzed by agarose gel electrophoresis. Then classify and summarize all samples according to different enzyme digestion results, and conclude different types.

[0044] There are three kinds of enzyme digestion results, namely complete mutation of ...

Embodiment 3

[0046] Example 3: Diagnostic application of molecular genetic markers prepared by the present invention in polymorphisms in different chicken populations

[0047] 1. Diagnosis in population polymorphism: Using the above-mentioned SNP polymorphism detection method, the plin1 gene of 616 Luqin No. 3 chickens, 1478 Luqin No. 1 chickens and 68 Shiqiza chickens were detected by specific enzyme digestion.

[0048] 2. Frequency statistical analysis of SNP loci:

[0049] Genotype frequency refers to the ratio of the number of individuals of a certain genotype to the total number of individuals in a population. Paa=Aaa / N, where Paa represents the AA genotype frequency at a certain locus, Aaa represents the number of individuals with the AA genotype in the population, and N is the total number of the detection population.

[0050] Gene frequency refers to the relative ratio of the number of a certain gene to the total number of alleles in a population. The calculation formula can be w...

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Abstract

The invention discloses an excellent slaughter trait molecular genetic marker of broiler chicken. The molecular genetic marker is a polymorphic site on a plin1 single nucleotide polymorphism sequence of broiler chicken; the site is a base polymorphic site with A or T on the 49th site of an eighth exon of the plin1 gene sequence, and shows three genotypes of AA, AT or TT; and the TT type is the excellent slaughter trait molecular genetic marker of broiler chicken. The invention further discloses an application of the marker in auxiliary screening or prediction of the broiler chicken variety with excellent slaughter traits. An experiment proves that the polymorphism of the specific restriction site is relevant to the production performance; the measurement indexes of the individual body of which the genotype is TT are higher than those of the individual bodies of which the genotypes are AA and AT; and the individual body of which the genotype is TT can be reminded to breed the broiler chicken with excellent slaughter traits as chicken.

Description

technical field [0001] The invention belongs to the field of molecular biology technology, and relates to a molecular genetic marker for excellent slaughtering traits of broiler chickens and its application in assisting screening or prediction of broiler chicken breeds with excellent slaughtering traits. Background technique [0002] DNA markers, also known as DNA polymorphism markers and DNA molecular markers, are a direct reflection of genetic polymorphisms at the DNA level and are also an important part of genetic markers. Compared with morphological markers, cytological markers and protein markers, DNA molecular markers have many advantages: not affected by environmental conditions, not limited by developmental stages, nor limited by individuals and biological tissues and organs; at the same time, genomic DNA variation is very rich , the number of molecular markers to choose from greatly exceeds the number of morphological markers, cytological markers and protein markers...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12Q1/68
Inventor 林浴霜王丹丹胡玮曹顶国周艳雷秋霞
Owner SHANDONG UNIV
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