Method for predicating milk yield of small tailed han sheep by virtue of BMPR (bone morphogenetic protein receptor )-1B or PRLR (prolactin receptor) gene SNP (single nucleotide polymorphism) lotus

A technology for small-tailed Han sheep and milk production, applied in the field of animal husbandry, can solve problems such as inability to predict the future or newly born livestock, time-consuming and labor-intensive, etc., achieve good application prospects, improve survival rate, improve survival rate and development speed Effect

Active Publication Date: 2014-10-08
新疆维吾尔自治区畜牧科学院中国-澳大利亚绵羊育种研究中心
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Problems solved by technology

However, the traditional method of measuring lactation is time-consuming and labor-intensiv...

Method used

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  • Method for predicating milk yield of small tailed han sheep by virtue of BMPR (bone morphogenetic protein receptor )-1B or PRLR (prolactin receptor) gene SNP (single nucleotide polymorphism) lotus
  • Method for predicating milk yield of small tailed han sheep by virtue of BMPR (bone morphogenetic protein receptor )-1B or PRLR (prolactin receptor) gene SNP (single nucleotide polymorphism) lotus
  • Method for predicating milk yield of small tailed han sheep by virtue of BMPR (bone morphogenetic protein receptor )-1B or PRLR (prolactin receptor) gene SNP (single nucleotide polymorphism) lotus

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Experimental program
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Effect test

Embodiment 1

[0021] Embodiment 1: BMPR-1B gene A746G site detection method

[0022] 1. Sample Collection and DNA Extraction

[0023] Blood was collected from the jugular vein of Small-tailed Han sheep, 2-5mL / sheep, anticoagulated with sodium heparin, and frozen at -20°C. Genomic DNA was extracted by phenol-chloroform extraction, dissolved in ultrapure water, and stored at 4°C until use.

[0024] 2. Primer sequence

[0025] According to the sequence of the sheep bone morphogenetic protein receptor IB (bone morphogenetic protein receptor IB, BMPR-IB) gene (GenBank accession numbers AF357007 and AF298885) containing the A746G mutation site, primers were designed, the amplification length was 140bp, and the primer sequence was:

[0026] Forward: 5'-GTCGCTATGGGGAAGTTTGGATG-3';

[0027] Reverse: 5'-CAAGATGTTTTCATGCCTCATCAACACGGTC-3'.

[0028] 3.PCR-RFLP

[0029] PCR 20μl reaction system composition: Taq DNA polymerase (5u / μl) 0.12μl; 10×PCR buffer 2μl; MgCl2 (25mmol / L) 2μl; dNTP (2.5mmol / L)...

Embodiment 2

[0034] Embodiment 2, PRLR gene E2-C34T site detection method

[0035] 1. Sample Collection and DNA Extraction

[0036] Blood was collected from the jugular vein of Small-tailed Han sheep, the blood sample was 2-5mL / sheep, anticoagulated with heparin sodium, -20. C frozen. Genomic DNA was extracted by phenol-chloroform extraction, dissolved in ultrapure water, and stored at 4°C until use.

[0037] 2. E2-C34T mutation site sequencing primer sequence

[0038] Primers were designed according to the second exon sequence of the sheep prolactin receptor gene PRLR, and the primer sequences were:

[0039] Forward: 5'-GACGTTTCTCTCCACCACAGAT-3'

[0040] Reverse: 5'-CCAACACACACGTGCTCAGA-3'

[0041] The length of the PCR product was 270bp, and the PCR product was sent to Shanghai Sangon Biological Co., Ltd. for sequence determination. Determine the SNP site according to the sequencing map.

[0042] 3. Determine the sequencing results based on the sequencing map, see figure 1 .

Embodiment 3

[0043] Example 3, BMPR-1B gene and PRLR gene SNP site and lactation volume correlation analysis

[0044] 1. Determination of lactation volume of Small Tail Han sheep

[0045] Under the same feeding conditions in the same sheep farm, 48 small-tailed Han sheep were sampled and measured once a week after lambing, and the continuous measurement was 8 times for 2 months. During the measurement, the ewes and lambs were separated for 24 hours. The milk was milked and isolated at 20.00 pm the night before, and three milkings were performed at 8.00 am, 14.00 noon, and 20.00 pm the next day, and were weighed and recorded. The three times were combined as the ewe’s lactation volume for one day, and the milk was weighed and fed back to the ewe. lamb.

[0046] 2. Correlation analysis between BMPR-1B gene A746G locus and lactation volume

[0047] Using SPSS13.0 statistical analysis software, the least square variance analysis was used to analyze the correlation between individuals with di...

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Abstract

The invention relates to the raise livestock field, and discloses a method for predicating milk yield of small tailed han sheep by virtue of BMPR(bone morphogenetic protein receptor )-1B or PRLR (prolactin receptor) gene SNP (single nucleotide polymorphism) lotus. The method comprises the following steps: extracting a small tailed han sheep genome DNA (deoxyribonucleic acid); amplifying a sequence of SNP lotus-containing BMPR-IB A746G or a sequence of SNP lotus-containing PRLR gene; judging an E2-C34T mutant gene type of the BMPR-IB A746G or PRLR gene. The method disclosed by the invention can be applied to carrying out feeding and management according to difference of milk secretion control gene types, and can be used for improving the milk secretion amount, guaranteeing that a lamb has enough milk intake, improving the survival rate and reducing the breeding economic loss. The method disclosed by the invention provides a molecular-level technical support for predicating and molecularly selecting the milk yield of small tailed han sheep; by applying the method disclosed by the invention, the survival rate and development rate of the lambs are greatly improved, so that better breeding economic benefits are produced, and therefore, the method has good application prospect.

Description

technical field [0001] The invention relates to the field of animal husbandry, in particular to a method for predicting the lactation volume of small-tailed Han sheep by SNP sites of BMPR-1B or PRLR genes. Background technique [0002] Small-tailed Han sheep is a breed of sheep with both meat and coat. It grows fast, matures early, has strong fecundity, stable performance genetics, and strong adaptability. It matures early, has multiple births, and has multiple lambs: it grows fast, has a large body and more meat, and has good meat quality: Small-tailed Han sheep 4 It can be fattened and slaughtered at the age of one month, and the annual slaughter rate is more than 400%. The weight can reach 50 kg at the age of 6 months, 100 kg at the age of one year, and 130-190 kg for adult sheep. The slaughter rate of one-year-old fattening sheep is 55.6%, and the net meat rate is 45.89%. [0003] Small-tailed Han sheep have very strong fecundity, and they can produce three times in two...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6888C12Q2600/156
Inventor 史洪才牛志刚李晓林袁燕张宁贺三刚张雪梅黄俊成
Owner 新疆维吾尔自治区畜牧科学院中国-澳大利亚绵羊育种研究中心
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