CN114752681B - SNP marker affecting wool length of merino sheep in alpine and application thereof - Google Patents
SNP marker affecting wool length of merino sheep in alpine and application thereof Download PDFInfo
- Publication number
- CN114752681B CN114752681B CN202210391580.6A CN202210391580A CN114752681B CN 114752681 B CN114752681 B CN 114752681B CN 202210391580 A CN202210391580 A CN 202210391580A CN 114752681 B CN114752681 B CN 114752681B
- Authority
- CN
- China
- Prior art keywords
- sheep
- merino
- alpine
- wool length
- genotype
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6844—Nucleic acid amplification reactions
- C12Q1/6858—Allele-specific amplification
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/124—Animal traits, i.e. production traits, including athletic performance or the like
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/156—Polymorphic or mutational markers
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/80—Food processing, e.g. use of renewable energies or variable speed drives in handling, conveying or stacking
- Y02P60/87—Re-use of by-products of food processing for fodder production
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Analytical Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biophysics (AREA)
- Immunology (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Biotechnology (AREA)
- Physics & Mathematics (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention belongs to the technical field of molecular genetics, and particularly relates to an SNP marker related to wool length of merino sheep in alpine and application thereof, wherein the SNP molecular marker is located at 6296384 nucleotide site T/C mutation on chromosome 3 of the international sheep reference genome oar_v4.0 version. The invention also relates to a specific primer pair for detecting the SNP molecular marker by utilizing the PCR technology, a kit containing the primer pair and a nucleotide polymorphism detection method. SNP locus detection is used for developing the early selection of the wool length of the merino sheep in the alpine, shortening the cultivation period and accelerating the cultivation process, and a technique for the early selection of the wool length of the merino sheep in the alpine is established, so that the breeding time of the merino sheep in the alpine with excellent properties is reduced, the breeding cost is reduced, and the merino sheep has high application value.
Description
Technical Field
The invention belongs to the technical field of molecular genetics, and particularly relates to SNP markers related to wool length of merino sheep in high mountain and application thereof.
Background
Wool is a layer of textile-value fiber which is covered on the surface of sheep body and is a derivative of skin. Wool is a main raw material of the wool spinning industry, and accounts for about 97% of the wool spinning raw material, and is mainly used for processing products such as clothing fabrics, knitting wool, blanket and the like. In wool production and breeding practice, wool characteristics are important economic characteristics, and mainly comprise a plurality of indexes such as wool length, wool yield, fiber diameter, bending degree, breaking strength, elongation, wool yield, wool shearing amount and the like, and are closely related to weaving products and economic benefits. As the population grows, the demand for wool has exceeded supply. With the change of market demands, the production of high-quality wool has become a main goal of goat breeding.
The alpine merino sheep is a new merino sheep variety which takes Gansu alpine merino sheep as a female parent and Australian merino sheep as a male parent, comprehensively utilizes modern advanced biotechnology and breeding technology, and is suitable for the alpine arid ecological region with the altitude of 2400-4070 m in the first example of the world after 20 years of breeding. Wool length is also an important index for determining economic value, and objective inspection of wool quality is increasingly emphasized in wool sales, and is more and more tightly combined with spinning performance, breeding and production.
Studies show that wool traits are affected by genetic factors and non-genetic factors, wherein functional genes play an important role in hair follicle development, wool growth and physical and chemical properties of wool. Compared with the traditional breeding method, the molecular marker assisted selection method has a plurality of advantages, such as obviously shortening the generation interval, improving the selection accuracy, advancing the selection time, and simultaneously having good selection effect on the characteristics of low genetic strength, the characteristics which are not represented in early stage, the characteristics which are difficult to measure in living bodies or have larger measurement difficulty and higher cost. However, if the molecular marker assisted selection method is to be successfully applied to the production practice of wool length traits, a key gene for regulating wool production traits or a molecular genetic marker linked with the key gene needs to be found. In recent years, genetic breeding experts of fine wool sheep at home and abroad are constantly dedicated to research on excavating candidate genes or molecular markers for controlling wool traits, but a molecular marker system for the fine wool traits is not established so far, and a large number of important gene resources for regulating wool traits are not effectively excavated and utilized yet.
In order to accelerate the development of wool industry, the selection of candidate genes related to wool length or molecular markers linked to QTL at the molecular level has become a prime condition for breeding workers to realize auxiliary selection. Wherein SNP is a molecular genetic marker proposed by the human genome research center scholarer of the American Massachusetts institute of technology, lander, in 1996, mainly refers to DNA sequence polymorphism caused by variation of single nucleotide at genome level. SNPs exhibit polymorphisms that involve only single base variation, in terms of transitions, transversions, insertions, deletions, and the like. The SNP molecular marker has the advantages of stable heredity, low mutation rate, convenient automatic detection and the like. Therefore, the molecular auxiliary marker genes closely linked with the wool length are searched, and the functional genes for regulating and controlling the wool length characters are screened, so that the method is a beneficial means for realizing the organic combination of the modern molecular breeding technology and the conventional breeding technology and improving the breeding economic efficiency.
The invention discovers an SNP molecular marker affecting wool length of mountain merino sheep, which is positioned at 6296384 base on chromosome 3 of the international sheep genome oar_v4.0 version, and the mutant base is T or C. When the SNP molecular marker base is T, the genotype is TT; when the SNP molecular marker base is C, the genotype is TC or CC; the wool length of mountain merino sheep of the genotype TT or TC is significantly greater than genotype CC (p < 0.05); the causative TT or TC showed no significant differences among individuals (p > 0.05). The method for detecting the nucleotide polymorphism related to the wool length of the merino sheep in the alpine region by using the PCR technology has the advantages of high accuracy, high detection speed, low cost and easier interpretation of the result. The method can realize automatic detection of SNP locus polymorphism related to wool length, can be used for breeding early selection and retention by detecting SNP loci of the wool length of the merino alpine, improves the breeding accuracy of the merino alpine, and has potential application value in large-scale molecular precise breeding of the merino alpine.
Disclosure of Invention
The invention provides an SNP molecular marker affecting the wool length of mountain merino sheep, and the genotyping of the wool length of mountain merino sheep is realized by detecting the base type of the SNP molecular marker, wherein the SNP molecular marker is positioned at the 6296384 base on chromosome 3 of the version of Oar_v4.0 of the international sheep genome, and the mutant base is T or C. When the SNP molecular marker base is T, the genotype is TT; when the SNP molecular marker base is C, the genotype is TC or CC; the wool length of mountain merino sheep of the genotype TT or TC is significantly greater than genotype CC (p < 0.05); genotype TT or TC individuals did not show significant differences (p > 0.05); and analyzing the wool length of the merino sheep in the mountain through genotyping, and carrying out breeding. The method specifically comprises the following steps:
in a first aspect, the invention provides an application of a reagent for detecting SNP molecular markers related to the wool length of high mountain merino sheep, wherein the SNP molecular markers are positioned at the base of the 6296384 site on chromosome 3 of the International sheep genome oar_v4.0 version 3; the mutant base is T or C.
Preferably, when the SNP molecular marker base is T, the genotype is TT; when the SNP molecular marker base is C, the genotype is TC or CC; the wool length of the mountain merino sheep of the genotype TT or TC is larger than that of the genotype CC.
Preferably, the reagents comprise a primer pair for amplifying a nucleotide sequence containing the SNP molecular marker.
Preferably, the nucleotide sequence of the SNP molecular marker is shown as SEQ ID NO.1, and the SNP molecular marker is positioned at the 250 th position.
In a second aspect, the invention provides an application of a reagent for detecting SNP molecular markers related to merino sheep wool length in the breeding of merino sheep in alpine, wherein the SNP molecular markers are positioned at the base of 6296384 locus on chromosome 3 of the international sheep genome oar_v4.0 version; the mutant base is T or C.
Preferably, when the SNP molecular marker base is T, the genotype is TT; when the SNP molecular marker base is C, the genotype is TC or CC; the wool length of the mountain merino sheep of the genotype TT or TC is larger than that of the genotype CC.
Preferably, the reagents comprise a primer pair for amplifying a nucleotide sequence containing the SNP molecular marker.
Preferably, the nucleotide sequence of the SNP molecular marker is shown as SEQ ID NO.1, and the SNP molecular marker is positioned at the 250 th position.
In a third aspect, the present invention provides a specific primer pair for amplifying the nucleotide sequence containing the SNP molecular marker according to the first or second aspect, wherein the primer pair has the sequence:
F：5'-GCCTGACTCGTCTCCTCTA-3'；
R：5'-CGTCCTCCTAATACTTTGCTC-3'。
in a fourth aspect, the invention provides the use of a specific primer pair as described in the third aspect for detecting the wool length of mountain merino sheep or for breeding mountain merino sheep.
Preferably, the method for realizing the detection of the wool length of the mountain merino sheep or the breeding of the mountain merino sheep comprises the following steps:
(1) Extracting blood genome DNA of the merino alpine sheep as a template DNA;
(2) Carrying out PCR amplification on the genomic DNA of the blood of the merino sheep to be detected obtained in the step (1) by using a specific primer pair to obtain a PCR amplification product;
(3) Purifying the PCR amplification product obtained in the step (2), and carrying out genotyping detection, wherein when the SNP molecular marker base is T, the genotype is TT; when the SNP molecular marker base is C, the genotype is TC or CC; the wool length of the mountain merino sheep of the genotype TT or TC is larger than that of the genotype CC.
Preferably, the specific primer pair sequences are:
F：5'-GCCTGACTCGTCTCCTCTA-3'；
R：5'-CGTCCTCCTAATACTTTGCTC-3'。
preferably, the PCR amplification system is 25 μl: 22. Mu.L of the premix, 1. Mu.L of each of the upstream and downstream primers, and 1. Mu.L of the template DNA.
Preferably, the PCR amplification procedure: 98 ℃ for 2min;98 ℃ for 10s,51 ℃ for 10s and 72 ℃ for 10s, and 35 cycles are total; extending at 72℃for 2min.
The beneficial effects of the invention are as follows: (1) the invention provides an SNP molecular marker affecting wool length of merino sheep in alpine, which is positioned at a base of 6296384 locus on chromosome 3 of the international sheep genome oar_v4.0 version 3; the mutation base is T or C; when the SNP molecular marker base is T, the genotype is TT; when the SNP molecular marker base is C, the genotype is TC or CC; the wool length of mountain merino sheep of the genotype TT or TC is significantly greater than genotype CC (p < 0.05); genotype TT or TC individuals did not show significant differences (p > 0.05); (2) the invention provides a method for detecting nucleotide polymorphism related to the wool length of mountain merino sheep by using a PCR technology, which has the advantages of high accuracy, high detection speed, low cost and easier interpretation of results. The method can realize automatic detection of SNP locus polymorphism of the merino sheep wool length in the alpine, can be used for early selection and retention of breeding and improves the breeding accuracy of the merino sheep in the alpine by detecting the SNP locus of the merino sheep wool length in the alpine, and has potential application value in large-scale molecular precise breeding of the merino sheep in the alpine.
Drawings
FIG. 1PCR amplification results;
the PCR product of FIG. 2 was purified and sequenced to obtain the genotype analysis result, wherein TT is TT genotype, TC is TC genotype, and CC is CC genotype.
Detailed Description
The following describes the technical scheme of the present invention in detail by referring to examples. It should be noted that the following examples are given for illustrative purposes only and are not intended to limit the scope of the present invention. Various modifications and alterations of this invention may be made by those skilled in the art without departing from the spirit and scope of this invention.
All the experimental methods of the experiments in the following examples are conventional methods unless otherwise specified.
The experimental conditions for all experiments in the examples described below are conventional, unless otherwise specified, such as the molecular cloning laboratory Manual of Sambrook et al (Sambrook J & Russell DW, molecular cloning: a laboratory manual, 2001), or as recommended by the manufacturer's instructions.
The SNP is short for single nucleotide polymorphism, and refers to DNA sequence polymorphism caused by single nucleotide variation at genome level.
Example 1 correlation between different genotypes of mountain merino sheep and wool length
1. Sample collection
The sample comes from a popularization station of the sheep breeding technology in Gansu province, 116 merino sheep blood samples with production records are collected, 5mL of each sheep vein blood is collected in a blood collection tube added with EDTA-K2 anticoagulant, the blood samples are quickly and uniformly mixed after being collected, the blood samples are put into a sampling box containing an ice bag for temporary storage, and the blood samples are transported back to a laboratory and are frozen and stored in a refrigerator at the temperature of minus 20 ℃ for DNA extraction. Each sheep wool length record is provided by a Gansu province sheep breeding technology popularization station.
2. Main reagent and instrument
EDTA-K2 vacuum blood collection tubes were purchased from Jiangsu Yuli medical instruments Co., ltd; blood genome extraction kit was purchased from tiangen biochemical technology (beijing) limited; nanoDrop2000 spectrophotometer us Thermo Fisher Scientific company; DL2000 Marker, agarose, nucleic acid dye were all purchased from Beijing Soy Bao technology Co., ltd; gold medal Mix (green) was purchased from beijing engine biotechnology limited; the electrophoresis apparatus is purchased from Beijing Liuyi instrument factory; PCR instrument was purchased from BioRad corporation.
3. Method of
3.1 extraction of genomic DNA from blood
Extracting genome DNA from blood sample by using blood genome extraction kit of Tiangen biochemical technology (Beijing) limited company, and detecting concentration and purity of the extracted DNA under ultraviolet spectrophotometer with concentration > 20 ng/. Mu. L, OD 260 /OD 280 The experimental requirements are met when the temperature is between 1.7 and 1.9, and the mixture is stored at the temperature of minus 20 ℃ for standby.
3.2 primer design
Referring to the international sheep genome oar_v4.0 version No. 3 chromosomal gene sequence (GenBank accession number: nc_ 019460.2), a pair of specific primers was designed using primer premier5.0 software, containing g6296384T > C SNP sites.
Primer sequence:
F：5'-GCCTGACTCGTCTCCTCTA-3'；
R：5'-CGTCCTCCTAATACTTTGCTC-3'。
the length of the amplified fragment is 371bp, and the primer is synthesized by Beijing qing biological science and technology Co.
3.3PCR amplification and sequencing
PCR amplification System 25. Mu.L: gold medal Mix (green) 22. Mu.L, 1. Mu.L each for the upstream and downstream primers, and 1. Mu.L for the template.
PCR amplification procedure: 98 ℃ for 2min;98 ℃ for 10s,51 ℃ for 10s and 72 ℃ for 10s, and 35 cycles are total; extending at 72℃for 2min.
And detecting the PCR product by using 1.5% agarose gel electrophoresis, and after the PCR product is qualified by using the agarose gel electrophoresis detection, sequencing by using a direct sequencing method, and completing sequencing by Beijing qingke biotechnology Co. The amplified nucleotide sequence is shown as SEQ ID No.1, and the SNP marker is positioned at 250 positions of the nucleotide sequence shown as SEQ ID No. 1.
And comparing the sequencing results of the PCR products by using biological analysis software Vector NTI advance 11.5.11.5, and analyzing a sequencing peak diagram to finish typing.
4. Statistical analysis
And counting the number of individuals with different genotypes at each site according to the genotyping result. The gene frequency, genotype frequency, effective allele factor (Ne), site heterozygosity (He) and Hardy-Weinberg equilibrium test of the SNP locus are calculated by using Popgen32 software, and the polymorphism information content is calculated by using PIC calculation software. The correlation of different genotypes of alpine merino sheep and wool length was analyzed using a general linear model in IBM SPSS Statistics software, and the results were expressed as "mean ± standard error".
5. Results
5.1PCR amplification and sequencing results
The SNP locus amplification product of the chromosome 3 of the merino sheep at mountain is detected by using 1.5% agarose gel, the result is shown in figure 1, the amplification product has clear band and good specificity, the size of the PCR product fragment is 371bp and accords with the expected size, and the next experiment can be carried out.
The peak pattern and sequence obtained after the PCR product is purified and sequenced are shown in FIG. 2. As can be seen from FIG. 2, the SNP site has T.fwdarw.C mutation, and three genotypes TT, TC and CC exist.
5.2 statistical analysis results
Genotype and allele frequency of the SNP locus of chromosome 3 of the merino alpine sheep were analyzed from the perspective of population genetics. As can be seen from Table 1, the TC genotype was most frequently represented as the dominant genotype, the C allele was 53% frequently represented as the dominant allele at the SNP site. The SNP site was shown to be in Hardy-Weinberg equilibrium (P > 0.05) by X2 fitness test (Table 1). The expected heterozygosity of the locus is 0.498, the polymorphism information content (polymorphism information content, PIC for short) is 0.374,0.25 < PIC < 0.50, and the locus belongs to moderate polymorphism.
TABLE 1 SNP site polymorphism of chromosome 3 g6296384T > C of mountain merino sheep
5.3 analysis of the association of different genotypes and lengths of Carnis Caprae Seu Ovis
The association of different genotypes and the hair length of the alpine merino sheep was analyzed by using a general linear model in IBM SPSS Statistics software, the hair length of the alpine merino sheep individuals with TT and TC genotypes was significantly longer than that of the alpine merino sheep individuals with CC genotypes (p < 0.05), and no significant difference was shown between the individuals with TT and TC genotypes (p > 0.05). The hair length of the alpine merino sheep can be judged by detecting the base of the SNP locus of the 3 rd chromosome of the alpine merino sheep. The results are shown in Table 2.
TABLE 2 correlation analysis between different genotypes and wool lengths of mountain merino sheep
Note that: the same row of data is marked with different lower case letters to indicate that the difference is significant (P < 0.05).
In summary, the SNP molecular marker is positioned at 6296384 bases on chromosome 3 of the international sheep reference genome oar_v4.0 version 3; the variation type is T/C, three genotypes exist, and when the 6296384 base on the 3 rd chromosome is T, the genotype is TT; when 6296384 bases on the chromosome 3 are C, the genotype is TC or CC; through correlation analysis of different genotypes and hair lengths, the hair lengths of the alpine merino sheep individuals with the TT and TC genotypes are found to be significantly longer than those of individuals with the CC genotype (p < 0.05), and no significant difference is shown between the individuals with the TT and TC genotypes (p > 0.05). The method can judge the wool length of the merino alpine through detecting the base of the 6296384 nucleotide locus on the chromosome 3 of the merino alpine, provides a basis for molecular marker assisted breeding of the merino alpine, can strengthen early selection of the merino alpine wool type, improve the seed selection accuracy, shorten the cultivation period and accelerate the cultivation process. Through the specific primer pair, a high-efficiency and accurate molecular marker assisted breeding technology can be established, the genetic progress of the merino wool length of the alpine merino can be increased by optimizing the dominant allele of the SNP molecular marker, and when the merino wool length fine character screening is carried out by adopting the molecular marker related to the merino wool length character, the merino sheep breeding method has the advantage of simple operation, can assist in screening the merino wool length, improves the precision of variety screening, reduces the breeding time of the merino wool length fine character of the alpine merino, reduces the breeding cost and increases the core competition.
Sequence listing
<110> China agricultural sciences Lanzhou livestock and veterinary drug institute
<120> SNP marker affecting wool length of merino sheep in mountain and application thereof
<160> 3
<170> SIPOSequenceListing 1.0
<210> 1
<211> 371
<212> DNA
<213> Artificial sequence (Artificial Sequence)
<400> 1
gcctgactcg tctcctctac gaacagggac agctgtcaca tccgtcatgc agggactgtg 60
cgagatgctg acagcatcct tggcacaggg ctggacataa aaaataatcg atggcaggac 120
ctttagtatc atgacctcag tgctctttta aatcagttca ccagctggct cctgcatgga 180
gcttggcaac tccagccagg acataccttc ctctgagaag gattcccagc acaaatgggg 240
gcagagagat ggatcagctt ttatcggcta ccagctgtta cctccgctca gccactgtct 300
gctgggagct ttgcaaacct gattctcttg ggacacccaa acaggcccat gagcaaagta 360
ttaggaggac g 371
<210> 2
<211> 19
<212> DNA
<213> Artificial sequence (Artificial Sequence)
<400> 2
gcctgactcg tctcctcta 19
<210> 3
<211> 21
<212> DNA
<213> Artificial sequence (Artificial Sequence)
<400> 3
cgtcctccta atactttgct c 21
Claims (7)
1. The application of a reagent for detecting SNP molecular markers related to the wool length of alpine merino sheep in detecting the wool length of alpine merino sheep is characterized in that the SNP molecular markers are positioned at the base of the 6296384 site on chromosome 3 of the international sheep genome oar_v4.0 version; the mutation base is T or C; the wool length of the mountain merino sheep of the locus genotype TT or TC is obviously larger than that of genotype CC.
2. The application of a reagent for detecting SNP molecular markers related to the wool length of the merino alpine sheep in the wool length trait breeding of the merino alpine sheep is characterized in that the SNP molecular markers are positioned at the base of the 6296384 locus on chromosome 3 of the international sheep genome oar_v4.0 version; the mutation base is T or C; the wool length of the mountain merino sheep of the locus genotype TT or TC is obviously larger than that of genotype CC; and selecting the high mountain merino sheep with genotype TT or TC for breeding.
3. The use according to claim 1 or 2, wherein the reagents comprise a primer pair for amplifying a nucleotide sequence containing the SNP molecular markers.
4. The use according to claim 3, wherein the nucleotide sequence containing the SNP molecular marker is shown as SEQ ID NO.1, the SNP molecular marker is positioned at 250 th position of the sequence shown as SEQ ID NO.1, and the primer pair has the sequence as follows:
F：5'-GCCTGACTCGTCTCCTCTA-3'；
R：5'-CGTCCTCCTAATACTTTGCTC-3'。
5. the application of a specific primer for amplifying SNP molecular markers related to the wool length of high mountain merino sheep in the wool length of high mountain merino sheep or the wool length trait breeding of high mountain merino sheep, wherein the SNP molecular markers are positioned at the base of 6296384 site on chromosome 3 of the international sheep genome oar_v4.0 version; the specific primer pair sequences are as follows:
F：5'-GCCTGACTCGTCTCCTCTA-3'；
R：5'-CGTCCTCCTAATACTTTGCTC-3'；
the method for detecting the wool length of the mountain merino sheep or the wool length character breeding of the mountain merino sheep comprises the following steps:
(1) Extracting blood genome DNA of the merino alpine sheep as a template DNA;
(2) Carrying out PCR amplification on the genomic DNA of the blood of the merino sheep to be detected obtained in the step (1) by using a specific primer pair to obtain a PCR amplification product; the specific primer pair sequences are as follows:
(3) Purifying the PCR amplified product obtained in the step (2), and carrying out genotyping detection, wherein the wool length of the alpine merino sheep with genotype TT or TC is obviously larger than that of the alpine merino sheep with genotype CC; and selecting the high mountain merino sheep with genotype TT or TC for breeding.
6. The use according to claim 5, wherein the PCR amplification system is 25 μl: 22. Mu.L of the premix, 1. Mu.L of each of the upstream and downstream primers, and 1. Mu.L of the template DNA.
7. The use of claim 5, wherein the PCR amplification procedure: 98 ℃ for 2min;98 ℃ for 10s,51 ℃ for 10s and 72 ℃ for 10s, and 35 cycles are total; extending at 72℃for 2min.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210391580.6A CN114752681B (en) | 2022-04-14 | 2022-04-14 | SNP marker affecting wool length of merino sheep in alpine and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210391580.6A CN114752681B (en) | 2022-04-14 | 2022-04-14 | SNP marker affecting wool length of merino sheep in alpine and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114752681A CN114752681A (en) | 2022-07-15 |
| CN114752681B true CN114752681B (en) | 2023-04-25 |
Family
ID=82331183
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210391580.6A Active CN114752681B (en) | 2022-04-14 | 2022-04-14 | SNP marker affecting wool length of merino sheep in alpine and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114752681B (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115896310B (en) * | 2022-12-09 | 2023-09-19 | 中国农业科学院兰州畜牧与兽药研究所 | Method for detecting CACNA1S gene CNV mark of mountain merino sheep and application thereof |
| CN115710603B (en) * | 2022-12-09 | 2023-08-22 | 中国农业科学院兰州畜牧与兽药研究所 | Method for detecting CNV (complementary factor v) mark of INPP5E gene of merino sheep in high mountain and application |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105886615A (en) * | 2016-04-20 | 2016-08-24 | 吉林省农业科学院 | Screening of SNP (Single Nucleotide Polymorphism) related to sheep wool traits and application |
| CN109055579A (en) * | 2018-10-24 | 2018-12-21 | 浙江省农业科学院 | Molecular labeling including SNP10-3 and its application in sheep assistant breeding |
| CN112921102A (en) * | 2021-03-11 | 2021-06-08 | 中国农业科学院北京畜牧兽医研究所 | SNP (Single nucleotide polymorphism) marker related to fine wool sheep wool character and detection primer group, kit, detection method and application thereof |
| CN114214428A (en) * | 2021-12-16 | 2022-03-22 | 中国农业科学院兰州畜牧与兽药研究所 | SNP molecular marker influencing mohair shearing amount of alpine merino and application thereof |
| CN114214426A (en) * | 2021-12-16 | 2022-03-22 | 中国农业科学院兰州畜牧与兽药研究所 | SNP molecular marker influencing alpine merino wool length character and application thereof |
-
2022
- 2022-04-14 CN CN202210391580.6A patent/CN114752681B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105886615A (en) * | 2016-04-20 | 2016-08-24 | 吉林省农业科学院 | Screening of SNP (Single Nucleotide Polymorphism) related to sheep wool traits and application |
| CN109055579A (en) * | 2018-10-24 | 2018-12-21 | 浙江省农业科学院 | Molecular labeling including SNP10-3 and its application in sheep assistant breeding |
| CN112921102A (en) * | 2021-03-11 | 2021-06-08 | 中国农业科学院北京畜牧兽医研究所 | SNP (Single nucleotide polymorphism) marker related to fine wool sheep wool character and detection primer group, kit, detection method and application thereof |
| CN114214428A (en) * | 2021-12-16 | 2022-03-22 | 中国农业科学院兰州畜牧与兽药研究所 | SNP molecular marker influencing mohair shearing amount of alpine merino and application thereof |
| CN114214426A (en) * | 2021-12-16 | 2022-03-22 | 中国农业科学院兰州畜牧与兽药研究所 | SNP molecular marker influencing alpine merino wool length character and application thereof |
Non-Patent Citations (5)
| Title |
|---|
| Hongchang Zhao等.Whole-genome re-sequencing association study on yearling wool traits in Chinese fine-wool sheep.Journal of Animal Science.2021,第99卷(第9期),第1-11页. * |
| P K Chitneedi等.Estimations of linkage disequilibrium, effective population size and ROH-based inbreeding coefficients in Spanish Churra sheep using imputed high-density SNP genotypes.Anim Genet .2017,第48卷(第4期),436-446. * |
| 乔国艳 等.不同数据结构和动物模型对高山美利奴羊经济性状遗传参数估计的比较.中国畜牧兽医.2020,(第02期),第531-543页. * |
| 张剑搏 等.不同动物模型对高山美利奴羊早期生长性状遗传参数估计的比较.中国农业科学.2018,(第06期),第1202-1212页. * |
| 陈来运 等.4个绵羊品种FSHR基因多态性与生物信息学分析.江苏农业科学.2019,(第22期),第47-51页. * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN114752681A (en) | 2022-07-15 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN113416790B (en) | SNP molecular marker influencing clean wool rate of alpine merino sheep and application thereof | |
| CN114214426B (en) | SNP molecular marker influencing alpine merino wool length traits and application thereof | |
| CN114752680B (en) | SNP marker affecting diameter of wool fibers of merino sheep in alpine and application thereof | |
| CN114214428B (en) | SNP molecular marker influencing mohair shearing amount of alpine merino and application thereof | |
| CN114752681B (en) | SNP marker affecting wool length of merino sheep in alpine and application thereof | |
| CN112048562B (en) | SNP molecular marker influencing diameter of alpine merino sheep wool fiber and application thereof | |
| CN113584183B (en) | SNP molecular marker influencing weaning weight traits of alpine merino sheep and application thereof | |
| CN114657266B (en) | SNP molecular marker for identifying shearing quantity of merino sheep in the whole year and application thereof | |
| CN112430669B (en) | SNP molecular marker influencing dispersion of diameter of alpine merino sheep wool and application thereof | |
| CN116083592A (en) | Molecular marker related to sheep growth traits and application thereof | |
| CN114657265B (en) | SNP marker for identifying weaning weight of alpine merino sheep and application thereof | |
| CN115029445B (en) | SNP marker related to weaning weight of merino sheep in alpine and application thereof | |
| CN113265473B (en) | SNP molecular marker influencing birth weight of alpine merino sheep and application thereof | |
| CN114250309A (en) | Molecular marker influencing diameter property of cashmere fiber of Xinjiang nan cashmere goat and specific primer pair and application thereof | |
| CN117721216A (en) | SNP molecular marker for identifying lambing number character of merino sheep for meat and application thereof | |
| CN116875706A (en) | SNP locus related to fine wool sheep net wool rate and application thereof | |
| CN117025787A (en) | SNP locus related to fine wool sheep net wool rate and application thereof | |
| CN116855616A (en) | SNP locus related to fine wool sheep net wool rate and application thereof | |
| CN116411127B (en) | Molecular marker primer combination for rapidly identifying mature-period characters of peach fruits and application thereof | |
| CN115927668B (en) | Method for detecting wool characters of merino sheep in mountain by using CCND1 gene CNV marker in auxiliary mode and application of method | |
| CN116121400B (en) | Wool character molecular marker assisted selection method based on alpine merino DLL1 gene CNV marker | |
| CN115851985B (en) | Method for auxiliary detection of wool characters by AMH gene CNV (complementary deoxyribonucleic acid) markers of merino sheep at high mountain and application | |
| CN117987569B (en) | Molecular marker related to immune traits and used as auxiliary selection of Tibetan sheep marker and application thereof | |
| CN115896310B (en) | Method for detecting CACNA1S gene CNV mark of mountain merino sheep and application thereof | |
| CN115747346B (en) | Method for detecting LMNA gene CNV mark of mountain merino sheep and application |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |