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Multi-gene polymerization effect analytic breeding method for increasing lambing number of goats

A litter size and multi-gene technology, which is applied in biochemical equipment and methods, microbial measurement/inspection, DNA/RNA fragments, etc., can solve the problems of long time-consuming, poor selection effect of single gene molecular markers, low breeding efficiency, etc.

Active Publication Date: 2018-12-07
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] The purpose of the present invention is to overcome the defects and deficiencies of the prior art in the genetic selection of litter size traits, to provide a multi-gene aggregation molecular marker that affects the litter size traits of goats and a multi-gene aggregation breeding method for improving the litter size traits of goats , to overcome the shortcomings of conventional breeding techniques such as long time-consuming, low breeding efficiency and poor selection effect of single gene molecular markers for complex economic traits

Method used

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  • Multi-gene polymerization effect analytic breeding method for increasing lambing number of goats
  • Multi-gene polymerization effect analytic breeding method for increasing lambing number of goats
  • Multi-gene polymerization effect analytic breeding method for increasing lambing number of goats

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0076] The establishment of the PCR-RFLP detection method of embodiment 1 GnRHR gene

[0077] (1) Primer design

[0078] According to the goat GnRHR gene sequence (Gene ID on NCBI: 100860755), utilize Primer5.0 software to design a pair of primers, sequence is as follows:

[0079] GnRHR:

[0080] SEQ ID NO.4: Upstream primer PCR-F: 5'-TCTTGAAGCTGTATCAGCCATA-3';

[0081] SEQ ID NO.5: Downstream primer PCR-R: 5'-GTGTTGAAAATTGTGGAGAGTAGA-3'.

[0082] (2) PCR amplification system and program setting

[0083] PCR reaction system: 10 μL system contains 1 μL of genomic DNA template, 0.2 μL of upstream and downstream primers, 5 μL of TaqDNA polymerase, supplemented with deionized double-distilled water to the final volume.

[0084] PCR reaction program: pre-denaturation at 94°C for 2 min, 35 cycles (denaturation at 94°C for 30 s, annealing at 56°C for 30 s, extension at 72°C for 30 s), extension at 72°C for 5 min, and storage at 4°C.

[0085] The PCR reaction product was detected...

Embodiment 2

[0102] The establishment of the PCR-HRM detection method of embodiment 2 FSHR gene

[0103] (1) Primer design for high resolution melting curve (HRM)

[0104] According to the goat FSHR gene sequence (Gene ID on NCBI: 100861291), utilize Primer5.0 software to design a pair of primers, sequence is as follows:

[0105] FSHR:

[0106] SEQ ID NO.6: Upstream primer PCR-F: 5'-TACCAGCTCCCAACGCAGAC-3';

[0107] SEQ ID NO.7: Downstream primer PCR-R: 5'-GACAGAGTCGATGGTGGCAT-3'.

[0108] (2) High resolution melting curve (HRM) reaction conditions

[0109] Use 55-65°C temperature gradient PCR to understand the optimal annealing temperature of the primers to obtain specific PCR products and improve the sensitivity of high-resolution melting.

[0110] PCR reaction system (10 μL): DNA template 1 μL, upstream and downstream primers 0.3 μL each, Dye 5 μL, RNase-free water 3.4 μL.

[0111] The PCR reaction program was: pre-denaturation at 94°C for 2 min, 40 cycles (denaturation at 94°C for...

Embodiment 3

[0129] The establishment of the PCR-RFLP detection method of embodiment 3 BMP6 gene

[0130] (1) Primer design

[0131] According to the goat BMP6 gene sequence (Gene ID on NCBI: 100860793), utilize Primer5.0 software to design a pair of primers, sequence is as follows:

[0132] BMP6:

[0133] SEQ ID NO.8: Upstream primer PCR-F: 5'-TGTGCCCTCACCTGCTGTCTC-3';

[0134] SEQ ID NO.9: Downstream primer PCR-R: 5'-CCCGGCCTTCCTCCTTTAACTC-3'.

[0135] (2) PCR amplification system and program setting

[0136] PCR reaction system: 10 μL system contains 1 μL of genomic DNA template, 0.2 μL of upstream and downstream primers, 5 μL of TaqDNA polymerase, supplemented with deionized double-distilled water to the final volume.

[0137] PCR reaction program: pre-denaturation at 94°C for 2 min, 35 cycles (denaturation at 94°C for 30 s, annealing at 59°C for 30 s, extension at 72°C for 30 s), extension at 72°C for 5 min, and storage at 4°C.

[0138] PCR reaction product is detected with 2% ag...

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Abstract

The invention belongs to the field of molecular biological technology and molecular marker breeding and especially relates to a multi-gene polymerization effect analytic breeding method for increasinglambing number of goats. In the method, GnRHR, FSHR and BMP6 genes are used as the candidate genes that influence the character of the lambing number of goats. In each of the three genes, there is apolymorphic site, so that by performing GnRHR-FSHR-BMP6 multi-gene polymerization effect model analysis to the molecular markers, it is found that AATTCC is an optimum combined genotype. By means of the optimization of combined genotype, the genetic progress on the character of lambing number of Chongqing Black goats is accelerated, and early-stage selection efficiency of the Chongqing Black goatsis increased, so that the method has important significance to cultivation of new species (lines) of high-quality high-reproductive goats.

Description

technical field [0001] The invention belongs to the technical fields of goat molecular biotechnology and molecular markers, and in particular relates to a breeding method for analyzing the effect of multigene aggregation for increasing the litter size of goats. In the present invention, the aggregation effect analysis of the molecular marker and the litter size of goats is applied to the early breeding work of the litter size traits of goats. Background technique [0002] At present, the degree of intensification of the goat industry is not high, most of them are free-range breeding, and the degree of improved breeds is low, the level of productivity is not high, and the economic benefits are not ideal. Therefore, the healthy and sustainable development of the sheep industry is a top priority. Reproductive traits are important economic traits. For breeding stock, fecundity is productivity, and reproductive performance is directly related to the economic effect of the mutton...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888C12Q2600/124C12Q2600/156
Inventor 李耀坤杨新月刘德武柳广斌孙宝丽
Owner SOUTH CHINA AGRI UNIV
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