Polygene pyramiding early-breeding method for raising litter size of sheep

A technology for lambing number and sheep, applied in the direction of DNA/RNA fragment, recombinant DNA technology, etc., can solve the problems of accelerating the process of breeding new products of high fecundity sheep, time-consuming trait measurement, long breeding cycle, etc. Accuracy, shortened breeding process, high workload effect

Inactive Publication Date: 2015-07-15
LANZHOU UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The present invention can solve the main defects of existing methods for cultivating new breeds of high-fecundity sheep, such as long breeding cycle, time-consuming and heavy workload for character determination, and accelerate the cultivation process of new breeds of high-fecundity sheep

Method used

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  • Polygene pyramiding early-breeding method for raising litter size of sheep
  • Polygene pyramiding early-breeding method for raising litter size of sheep
  • Polygene pyramiding early-breeding method for raising litter size of sheep

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] Embodiment 1, establishment of FecB gene PCR-RFLP diagnostic method

[0060] (1) Primer design

[0061] According to the sheep FecB gene sequence (GenBank accession number: NC_019463.1), a pair of primers M-F and M-R were designed using Primer5.0 software, the sequence is as follows:

[0062] FecB: M-F: 5'-GTCGCTATGGGGAAGTTTGGATG-3',

[0063] M-R: 5'-CAAGATGTTTTCATGCCTCATCAACACGGTC-3'.

[0064] (2) PCR amplification conditions

[0065] The total volume of the PCR reaction is 20 μL, including about 100 ng of sheep genomic DNA, containing 1 times the buffer (purchased from Promega Company), 1.5 mmol / L MgCl 2 , the final concentration of dNTP is 150 μmol / L, the final concentration of primer is 0.4 μmol / L, 2U Taq DNA polymerase (Promega). The PCR amplification program was: 94°C for 3min, 35 cycles of 94°C for 30s, 59°C for 30s, then 72°C for 25s, and finally 72°C for 5min. PCR reaction products were detected by 2% agarose gel electrophoresis. Obtain a 140bp specific a...

Embodiment 2

[0076] Embodiment 2, the establishment of BMP-15 gene PCR-RFLP diagnostic method

[0077] (1) Primer design

[0078] According to the sheep BMP-15 gene sequence (GenBank accession number: NC_019484.1), utilize Primer5.0 software to design a pair of primers M-F and M-R, the sequence is as follows

[0079] BMP-15: M-F: 5'-CACTGTCTTCTTGTTACTGTATTTCAATGAGAC-3',

[0080] M-R: 5'-GATGCAATACTGCCTGCTTG-3'.

[0081] (2) PCR amplification conditions

[0082] The total volume of the PCR reaction is 20 μL, including about 100 ng of sheep genomic DNA, containing 1 times the buffer (purchased from Promega Company), 1.5 mmol / L MgCl 2 , the final concentration of dNTP is 150 μmol / L, the final concentration of primer is 0.4 μmol / L, 2U Taq DNA polymerase (Promega). The PCR amplification program was: 94°C for 3min, 35 cycles of 94°C for 30s, 63°C for 30s, then 72°C for 25s, and finally 72°C for 5min. PCR reaction products were detected by 2% agarose gel electrophoresis. A 141bp specific am...

Embodiment 3

[0093] Embodiment 3, the establishment of FSHR gene PCR-RFLP diagnostic method

[0094] (1) Primer design

[0095] According to the sheep FSHR gene sequence (GenBank accession number: NC_019460.1), use Primer5.0 software to design a pair of primers M-F and M-R, the sequence is as follows

[0096] FSHR: M-F: 5'-CGTATCTTTTCCACGCCCTCT-3',

[0097] M-R: 5'-CCATCCACCCGATTGCTT-3'.

[0098] (2) PCR amplification conditions

[0099] The total volume of the PCR reaction is 20 μL, including about 100 ng of sheep genomic DNA, containing 1 times the buffer (purchased from Promega Company), 1.5 mmol / L MgCl 2 , the final concentration of dNTP is 150 μmol / L, the final concentration of primer is 0.4 μmol / L, 2U Taq DNA polymerase (Promega). The PCR amplification program was: 94°C for 3min, 35 cycles of 94°C for 30s, 58°C for 30s, then 72°C for 25s, and finally 72°C for 5min. PCR reaction products were detected by 2% agarose gel electrophoresis. Obtain 244bp specific amplification fragmen...

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Abstract

The invention belongs to the field of molecular breeding of livestock, and relates to a polygene pyramiding early-breeding method for raising the litter size of sheep. Cloning is performed from FecB, BMP15 and FSHR genes so as to obtain three associated gene segments, and the nucleotide sequences of the gene segments are shown in SEQ ID NO: 1-3. A base substitution of A111-G111 exists at the 111(th) bp of SEQ ID NO: 1, which causes the digestion polymorphism of Ava II-RFLP; a base substitution of C35-T35 exists at the 35(th) bp of the SEQ ID NO: 2, which causes the digestion polymorphism of Hinf I-RFLP; and a base substitution of C88-T88 exists at the 88(th) bp of the SEQ ID NO: 3, which causes the digestion polymorphism of BsiE I-RFLP. The molecular marker combination can significantly improve the polygene pyramiding early-breeding efficiency for raising the litter size of sheep.

Description

technical field [0001] The invention belongs to the technical field of sheep molecular breeding, and in particular relates to a method for early seed selection by multi-gene polymerization for increasing lamb size of sheep. The invention obtains molecular markers related to lamb size traits of sheep by cloning. Application of multigene aggregation in early breeding selection for improving sheep litter size by using the molecular markers in combination. The molecular markers are cloned from FecB, BMP15 and FSHR genes. Background technique [0002] According to the statistics of FAO (2012), there are 187 million sheep in my country, accounting for 16% of the world's breeding (1.169 billion), ranking first in the world (http: / / www.fao.org / home / en / ), It is the largest producer and consumer of mutton. However, there is still a certain gap between the overall production level of my country's sheep industry and foreign developed countries. Since 1970, developed countries in the s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12Q1/68
Inventor 李发弟王维民刘世佳喇永富潘香羽李冲翁秀秀马友记唐德富
Owner LANZHOU UNIVERSITY
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