Molecular marker related to character of milk yield of milk goat and application thereof

A technology of molecular markers and dairy goats, which is applied in the detection/testing of microorganisms, DNA/RNA fragments, recombinant DNA technology, etc., to achieve the effects of shortening the generation interval, high genotyping efficiency, and reducing breeding costs

Inactive Publication Date: 2017-05-31
青岛市畜牧兽医研究所
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In recent years, scholars at home and abroad have conducted some research on the genes related to milk production traits of dairy goats, and found some functional genes and molecular markers, but there are few research reports on milk production traits

Method used

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  • Molecular marker related to character of milk yield of milk goat and application thereof
  • Molecular marker related to character of milk yield of milk goat and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Example 1 Amplification of Partial Sequence of H-FABP Gene of Laoshan Dairy Goat

[0054] 1. Primer design

[0055] Using the online primer design software Primer-BLAST (https: / / www.ncbi.nlm.nih.gov / tools / primer-blast / ), the goat H-FABP gene sequence (GeneBank accession number: NC_030809.1, sequence interval 13664760- 13673332) was used as a template to design amplification primers containing partial sequences. The primers were synthesized by Qingdao Qingke Zixi Biotechnology Co., Ltd. The DNA sequences of the primer pairs are as follows:

[0056] H-FABPF5:TGTTCTGACCCCAAACTCGG

[0057] H-FABPR5:AGGGCTAGAGAACTGCTCCG

[0058] 2. Purification and sequencing of PCR products

[0059] PCR amplification reaction system 25 μL, including 1 μL of dairy goat peripheral blood genomic DNA (50ng / μL), 12.5 μL of 2×PCR Mix (Shanghai Sangon Bioengineering Co., Ltd.), 0.75 μL of forward primer H-FABPF4 (10mM ), 0.75 μL of reverse primer H-FABPR4 (10 mM), 10 μL of ddH 2 O. The react...

Embodiment 2

[0071] Example 2 Genotyping

[0072] The M of the 147th mutation site in the above sequence is A or C, and the mutation leads to SmaI-RFLP polymorphism. However, there is also a SmaI restriction site (CCCGGG, 165-170) near the mutation site at position 147 in this sequence, which makes it difficult to classify by restriction enzyme digestion, so another pair of primers (one of which is a mutation primer) was designed to solve the problem. For this difficulty, the sequence is as follows:

[0073] Sma1F:GGGACCAGGGGACGGATGTCG

[0074] Sma1R1:GCACCCAGTTTGCCTACGTCACCGCGGGTACCGGGCTCCGAGTCCCTGC

[0075] The sequence of the 176bp PCR product obtained by using the primer pair Sma1F and Sma1R1 amplified is as follows (the product was detected by electrophoresis on a 2% agarose gel, and the results are shown in figure 1 ):

[0076] GGGACCAGGGGACGGATGTCGGTGGCCTGCGCCCCAGCTCGCGGGCACCGTGCCCCCAAC

[0077] TGCTTCCTTTCCAGATTCCGAAGAGCGCCTTGAGGCCAGGGAAGGGAGCAGGGCTTACCA

[0078] GCCMGGGGCAGG...

Embodiment 3

[0080] Example 3 Association analysis of genotypes and traits of Laoshan dairy goat population

[0081] Select 200 Laoshan dairy goat ewes in lactation stage from Qingdao Aote sheep farm, collect whole blood from the jugular vein, extract genomic DNA, use primer pair Sma1F, Sma1R1, carry out PCR amplification on dairy goat population DNA, and use SmaⅠ enzyme digestion The genotype of each individual was detected, and it was found that there were 117 CC genotype individuals (group 1), and the average daily milk production measured was 2.32±0.68kg; the AC genotype individuals were 80 (group 2), and the daily milk production The average weight was 2.17±0.53kg; there were 3 individuals with AA genotype, because the number of individuals with AA genotype was too small to be statistically significant, so statistical analysis was not performed. Statistical analysis was performed on the data of group 1 and group 2 using one-way analysis of variance, and the obtained P value was 0.029 ...

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Abstract

The invention belongs to the technical field of animal molecular marker preparation and specifically relates to a molecular marker related to the character of milk yield of milk goat and an application thereof. The molecular marker is acquired in the manner of cloning a goat H-FABP gene; the sequence table of the nucleotide sequence is shown as SEQ ID NO.1; an A/C mutation is located at the No.124 locus basic group shown as the sequence table SEQ ID NO.1; Sma I-PFLP polymorphism is caused by the mutation. The invention also discloses a preparation method for the molecular marker and an application of a polymorphism detection method. A new molecular marker is supplied for the marker auxiliary breeding of the milk goat.

Description

technical field [0001] The invention belongs to the technical field of animal molecular marker preparation, and in particular relates to the preparation and application of a molecular marker related to milk production traits of dairy goats. Background technique [0002] my country is a country with a large number of dairy goats and a wide distribution. The dairy goat breeding industry used to be the leading industry of my country's dairy industry in the 1960s and 1980s. After the 1990s, it was affected by factors such as small breeding scale and rapid expansion of the dairy industry. The impact of dairy goat production slowed down. Over the past ten years, with the continuous improvement of people's living standards and the need for improved dietary structure, people's recognition of the nutritional value and safety of goat milk has changed, especially machine milking has solved the bottleneck problem of large-scale breeding , Promoted the revival and development of the nati...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6888C12Q2600/124C12Q2600/156
Inventor 李和刚程明宋晓娜杨峰郝小静张宝珣李培培杨培培包汉勋刘开东苗刚张明郭成玉
Owner 青岛市畜牧兽医研究所
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