CN105603092B - SNP marker relevant to the polygonal character of sheep and its application - Google Patents
SNP marker relevant to the polygonal character of sheep and its application Download PDFInfo
- Publication number
- CN105603092B CN105603092B CN201610082724.4A CN201610082724A CN105603092B CN 105603092 B CN105603092 B CN 105603092B CN 201610082724 A CN201610082724 A CN 201610082724A CN 105603092 B CN105603092 B CN 105603092B
- Authority
- CN
- China
- Prior art keywords
- sheep
- polygonal
- snp marker
- character
- primer
- 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
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
- 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
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Analytical Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Immunology (AREA)
- Molecular Biology (AREA)
- Biotechnology (AREA)
- Biophysics (AREA)
- Physics & Mathematics (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The present invention relates to SNP marker relevant to the polygonal character of sheep, the SNP marker is located at sheep MTX2 downstream of gene 53449bp, at the 344bp in sequence shown in SEQ ID NO.1, is mutated for G/A.The present invention also provides application of the SNP marker in the identification polygonal character of sheep, molecular genetic marker provided by the invention is not limited by the age of sheep, gender etc., early screening can be carried out to sheep to be measured, the problem of excellent polygonal kind of sheep selection time length in actual production is effectively relieved, lower breeding to spend, selecting and remain for the polygonal sheep individual in actual production is effectively increased, is increased economic efficiency.
Description
Technical field
The invention belongs to genetic biology fields, and in particular to SNP marker relevant to the polygonal character of sheep and its application.
Background technique
China has sheep variety resource abundant, and polygonal sheep is the sheep genetic resources of one type preciousness, research
Sheep with two or more angles on head is defined as polygonal sheep by person, wherein it is in the majority with the phenotype at four angles, also make quadrangle continuous
Sheep, it is existed only in a small number of ancient sheep varieties, and many polygonal sheep varieties have been in Critical Condition, and each state all carries out
Relevant conservation and breeding work establish polygonal sheep protection association such as to promote conservation, breeding and the business of polygonal sheep
Exchange.At present Europe, America polygonal sheep variety mainly include Icelandic sheep, Manx Logthan sheep and
Hebridean sheep etc..Also there are four polygonal sheep varieties, including Sishui fur sheep, Altai Sheep, Mongolian sheep, Ba Shibai in China
Sheep.Due to the inconvenience in production management, so that ratio of the polygonal sheep individual in these kinds is very low, some is on the verge of to go out
Absolutely.On the other hand, due to these polygonal sheep ornamental values with higher, in recent years, as non-traditional sheep variety, it is more
Angle sheep variety is cultivated and basic research is increasingly valued by people.
The polygonal character of sheep is a kind of inhereditary feature of preciousness, and the phenotype of sheep horn will be grown to half years old to 1 years old to it
Just it can determine whether.How quick and precisely to identify that the polygonal phenotype of sheep is to carry out the pass of polygonal Sheep Breeding using marker assisted selection
Key.Alderson etc. thinks that the heredity at angle is regulated and controled by two sites, and when two sites are all recessive homozygous, then sheep is more
Angle phenotype.Icelandic sheep studies have shown that in polygonal character and two corners character in this pair of of phenotype, polygonal is dominant something lost
It passes.To two polygonal sheep varieties of foreign countries of Jacob sheep and Navajo-Churro sheep studies have shown that being contaminated at sheep No. 2
There are the association sites of polygonal phenotype at colour solid 132Mb.Full genome is carried out to China's Mongolian sheep, Altai Sheep and Sishui fur sheep
Association analysis discovery, on No. 2 chromosomes of sheep at 132.6Mb, exists and the significant relevant SNP marker of the polygonal character of sheep.
Molecular breeding, i.e. molecular marker assisted selection breeding are referred to and are selected using DNA molecular marker breeding material
It selects, comprehensive improvement livestock and poultry important economical trait, is traditional genetic breeding and the breeding method that modern molecular biology organically combines.
With the fast development of molecular biology, molecular labeling in cattle breeding using increasingly extensive, importance is also increasingly convex
It is existing.Polygonal Sheep Breeding mostly uses traditional breeding way, and the determination of diagonal phenotype, the quantity at long angle to angle determines since lamb
Wait 6 months -12 months, breeding process is slow, and breeder wishes the DNA marker of selection with polygonal character close linkage, with reality
Now early stage chooses seeds and improves the target of breeding accuracy, to accelerate genetic breeding process.
Summary of the invention
The first purpose of the invention is to provide the SNP markers and its application of a kind of auxiliary identification polygonal character of sheep.
It is a further object to provide the primer for detecting SNP marker relevant to the polygonal character of sheep and contain
There is the kit of the primer.
In order to realize the first purpose of this invention, inventor has carried out greatly the sheep individual from different sheep varieties
The genotype of amount and polygonal phenotype correlation research, are found surprisingly that, in all polygonal sheep individuals, there is up to 96.9%
Individual the place MTX2 downstream of gene 53449bp for AA AG genotype.In the biangulate sheep individual of institute, have up to
It is GG genotype at the MTX2 downstream of gene 53449bp of 86.3% individual.Based on the above discovery, the present invention provides it is a kind of with
The relevant SNP marker of the polygonal character of sheep, the SNP marker are located at sheep MTX2 downstream of gene 53449bp, correspond to SEQ
At 344bp in sequence shown in ID NO.1, it is mutated for G/A.
The present invention also provides the specific primers for detecting SNP marker of the present invention, comprising:
Forward primer: 5 '-CCATTGCTGCCTGTTGTTTCT-3 '；
Reverse primer: 5 '-AGAATGATTTAGGAGGTGCGCT-3 '.
Application of the specific primer in the identification polygonal character of sheep also belongs to protection scope of the present invention.
It is the nucleotide as shown in SEQ ID NO.1 using the amplified production segment that the primer carries out PCR amplification acquisition
Sequence.
The present invention also provides application of the SNP marker in the identification polygonal character of sheep, comprising the following steps:
1) genomic DNA of sheep to be measured is extracted；
2) using the genomic DNA of sheep to be measured as template, pcr amplification reaction is carried out using specific primer, is expanded
Product segment；
3) pcr amplification product segment is detected, if the base at the 344bp of amplified production segment is A, sheep to be measured
For polygonal phenotypic advantage individual.
Wherein, the specific primer includes:
Forward primer: 5 '-CCATTGCTGCCTGTTGTTTCT-3 '；
Reverse primer: 5 '-AGAATGATTTAGGAGGTGCGCT-3 '.
Wherein, what the polygonal phenotypic advantage individual referred to is that sheep to be measured has up to 96.9% a possibility that for more
The sheep individual of angle character.
If at the 344bp of amplified production segment being GG genotype, it is two that sheep to be measured, which has 86.3% a possibility that,
The sheep at angle.
Wherein, the nucleotide sequence of the amplified production segment is as shown in SEQ ID NO.1.
The present invention is not particularly limited the method for detecting pcr amplification product segment, can use this field routine
Detection method carry out, for example, can be detected by gene order surveying method.
Wherein, the amplification system that PCR reaction uses in step 2) is calculated as with 40 μ l, 2 × Taq PCR MasterMix20 μ
L, 10pmol/ μ l forward primer and reverse primer each 0.5 μ l, 50ng/ μ l template DNA 1 μ l, ddH2O 18μl。
Wherein, in step 2) PCR react condition are as follows: 95 DEG C initial denaturation 5 minutes；95 DEG C are denaturalized 30 seconds, 60 DEG C of annealing 30
Second, 72 DEG C extend 45 seconds, and totally 35 recycle；Extend 5 minutes after 72 DEG C.
The present invention also provides the kits of the auxiliary identification polygonal character of sheep containing the specific primer.
Preferably, the kit further includes dNTPs, Taq archaeal dna polymerase, Mg2+, one of PCR reaction buffer
Or it is a variety of.
Preferably, the kit further includes standard positive template.
Specific primer of the present invention can also be combined with other for the specific primer of sheep Phenotypic examination
For sheep classification and breeding research.
Application of the SNP marker relevant to the polygonal character of sheep in polygonal sheep molecular mark also belongs to this
The protection scope of invention.
The present invention can carry out Genotyping to sheep using the SNP marker at sheep MTX2 downstream of gene 53449bp, and
The SNP marker and the polygonal phenotype of sheep are associated, the detection in the site, new material is not only provided for molecular breeding, but also
Marker assisted selection for the polygonal character of sheep provides scientific basis.
The present invention examines the genotype of sheep to be measured using polymerase chain reaction (PCR) and sequencing technologies, and advantage exists
In: molecular genetic marker provided by the invention is not limited by the age of sheep, gender etc., can carry out early stage sieve to sheep to be measured
Choosing is effectively relieved the problem of excellent polygonal kind of sheep selection time length in actual production, lowers breeding and spend, effectively increase practical life
In production polygonal sheep individual select and remain, increase economic efficiency, will polygonal Sheep Breeding work in play a great role.This hair
Bright identification method is easy to operate, expense is not high, accuracy is high, it can be achieved that automatic detection, thus has very high breeding real
Trample application value.The present invention can make have the accuracy identified in angle sheep individual up to 96.9%, to obtain considerable economy
Benefit.
Detailed description of the invention
Fig. 1 is the sequencing peak figure of three kinds of genotype of sheep of the present invention.
Specific embodiment
The present invention is described in detail With reference to embodiment.
The following examples are used to illustrate the present invention, but are not intended to limit the scope of the present invention..Unless otherwise specified, embodiment
In the conventional means that are well known to those skilled in the art of technological means used, raw materials used is commercial goods.
Embodiment 1
1, the acquisition of SNP marker relevant to the polygonal character of sheep
According to the information of sheep genomic DNA sequence, it is as follows to design pair of primers: forward primer F 5 '-
5 '-AGAATGATTTAGGAGGTGCGCT-3 ' of CCATTGCTGCCTGTTGTTTCT-3 ' and reverse primer R.It amplifies to be measured
Nucleotide fragments where SNP, as shown in SEQ ID NO.1.The SNP marker is located at the 344bp of the pcr amplified fragment, should
Locating base can be A or G.
Wherein PCR reaction system is calculated as with 40 μ l, 2 × Taq PCR MasterMix (comprising Taq archaeal dna polymerase,
dNTPs、MgCl2, reaction buffer, PCR reaction reinforcing agent and optimization agent and stabilizer) 20 μ l, 10pmol/ μ l primers Fs
With R each 0.5 μ l, 50ng/ μ l template DNA 1 μ l, ddH2O 18μl。
In aforementioned applications PCR react condition are as follows: 95 DEG C initial denaturation 5 minutes；95 DEG C are denaturalized 30 seconds, and 60 DEG C are annealed 30 seconds,
72 DEG C extend 45 seconds, 35 circulations；Extend 5 minutes after 72 DEG C.
2, PCR product sequencing and genotype determine
The agarose gel electrophoresis that pcr amplification product concentration is 1.5%, GoldView are dyed, in gel imaging system
PCR amplification effect is observed, forward primer 5 '-CCATTGCTGCCTGTTGTTTCT-3 ' and reverse primer 5 '-are utilized
AGAATGATTTAGGAGGTGCGCT-3 ' carries out sequencing (the limited public affairs of Aomei De-Nol (Beijing) Gene science to target fragment
Department), sequencing result shows that there are three kinds of genotype, i.e. AA, AG and GG (as shown in Figure 1) at the 344bp of target fragment.
The association analysis of embodiment 2 sheep type and polygonal phenotype
Using the method established in embodiment 1, PCR amplification and sequencing are carried out to the genomic DNA of 6 sheep varieties, surveyed
Determine Mongolian sheep, Altai Sheep, Sishui fur sheep, tibetan sheep, Suffolk and Sumatera sheep and amounts to 236 individuals, each kind
Sample number and angle phenotype are as shown in table 1, the genotype and phenotypic analysis result in the site pcr amplification product segment 344bp such as 2 institute of table
Show.
The sample number of 16 kind sheep of table and the phenotype at angle
Distribution of the genotype of SNP in 236 sheep individuals at 2 MTX2 downstream of gene 53449bp of table
The result shows that: in all polygonal individuals, 96.9% individual is AA AG genotype, i.e., in Sheep Populations
Judge that the accuracy rate of polygonal phenotype reaches 96.9% using the SNP；In the biangulate sheep individual of institute, 86.3% individual is
GG phenotype may determine that the accuracy rate of two corners individual reaches 86.3% using the site.
Although above the present invention is described in detail with a general description of the specific embodiments,
On the basis of the present invention, it can be made some modifications or improvements, this will be apparent to those skilled in the art.Cause
This, these modifications or improvements, fall within the scope of the claimed invention without departing from theon the basis of the spirit of the present invention.
Claims (9)
1. SNP marker relevant to the polygonal character of sheep, which is characterized in that the SNP marker is located at sheep MTX2 downstream of gene
At 53499bp, at the 344bp in sequence shown in SEQ ID NO.1, it is mutated for G/A.
2. the specific primer for detecting SNP marker described in claim 1 characterized by comprising
Forward primer: 5 '-CCATTGCTGCCTGTTGTTTCT-3 '；
Reverse primer: 5 '-AGAATGATTTAGGAGGTGCGCT-3 '.
3. application of the SNP marker described in claim 1 in the identification polygonal character of sheep, comprising the following steps:
1) genomic DNA of sheep to be measured is extracted；
2) using the genomic DNA of sheep to be measured as template, pcr amplification reaction is carried out using specific primer, obtains amplified production
Segment；
3) pcr amplification product segment is detected, if the base at the 344bp of amplified production segment is A, sheep to be measured is more
Angle phenotypic advantage individual；
Wherein, the specific primer includes:
Forward primer: 5 '-CCATTGCTGCCTGTTGTTTCT-3 '；
Reverse primer: 5 '-AGAATGATTTAGGAGGTGCGCT-3 '.
4. application according to claim 3, which is characterized in that the amplification system that PCR reaction uses in step 2) is with 40 μ l
It is calculated as, 2 × Taq PCR MasterMix20 μ l, 10pmol/ μ l forward primer and each 0.5 μ l, 50ng/ μ l template of reverse primer
DNA1 μ l, ddH2O18μl。
5. application according to claim 4, which is characterized in that the condition that PCR reacts in step 2) are as follows: 95 DEG C of initial denaturations 5
Minute；95 DEG C are denaturalized 30 seconds, and 60 DEG C are annealed 30 seconds, and 72 DEG C extend 45 seconds, and totally 35 recycle；Extend 5 minutes after 72 DEG C.
6. the kit of the auxiliary identification polygonal character of sheep containing specific primer described in claim 2.
7. kit according to claim 6, which is characterized in that the kit further includes dNTPs, Taq DNA polymerization
Enzyme, Mg2+, one of PCR reaction buffer or a variety of.
8. kit according to claim 6 or 7, which is characterized in that the kit further includes standard positive template.
9. SNP marker relevant to the polygonal character of sheep is in polygonal sheep molecular mark described in claim 1
Using.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610082724.4A CN105603092B (en) | 2016-02-05 | 2016-02-05 | SNP marker relevant to the polygonal character of sheep and its application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610082724.4A CN105603092B (en) | 2016-02-05 | 2016-02-05 | SNP marker relevant to the polygonal character of sheep and its application |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105603092A CN105603092A (en) | 2016-05-25 |
| CN105603092B true CN105603092B (en) | 2019-03-26 |
Family
ID=55983427
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610082724.4A Active CN105603092B (en) | 2016-02-05 | 2016-02-05 | SNP marker relevant to the polygonal character of sheep and its application |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105603092B (en) |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105950742B (en) * | 2016-05-31 | 2019-07-19 | 中国农业科学院北京畜牧兽医研究所 | A method of identification ovine growth character is assisted based on rs417014745 SNP site |
| CN113278714B (en) * | 2021-07-23 | 2021-11-09 | 中国农业大学 | Gene chip for analyzing whether sheep has horns or not, molecular probe combination, kit and application |
| CN117051131B (en) * | 2023-10-11 | 2024-01-30 | 中国农业科学院北京畜牧兽医研究所 | SNP molecular marker related to sheep brucellosis resistance character, detection primer and application thereof |
| CN118374609B (en) * | 2024-06-21 | 2024-08-30 | 中国农业科学院北京畜牧兽医研究所 | SNP molecular marker related to sheep growth traits and application thereof |
-
2016
- 2016-02-05 CN CN201610082724.4A patent/CN105603092B/en active Active
Non-Patent Citations (3)
| Title |
|---|
| 9个绵羊群体遗传多样性的微卫星分析;常爽;《河南农业大学硕士学位论文》;20101231;全文 * |
| Genome-wide association reveals the locus responsible for four-horned ruminant;James W. Kijas et al;《Animal Genetics》;20160114;第47卷(第2期);摘要 * |
| 基于cytb 基因探讨家绵羊和多角绵羊的系统发育;赵倩君等;《中国农业科学》;20101231;第43卷(第14期);第3005-3011页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN105603092A (en) | 2016-05-25 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105603092B (en) | SNP marker relevant to the polygonal character of sheep and its application | |
| CN105886642B (en) | SNP marker relevant to Chinese Sheep tail type character and its application | |
| WO2023001211A1 (en) | Gene chip, molecular probe combination and kit for analyzing traits of sheep wool, and use | |
| CN107326077A (en) | A kind of molecular labeling for differentiating spotted maigre genetic sex and its application | |
| CN113913533B (en) | SNP molecular marker related to grass carp characters and application thereof | |
| CN104561306B (en) | A kind of pig UNC13B gene SNP markers related with pig eye muscle area character | |
| CN105274094B (en) | SNP marker and its application | |
| CN104419706B (en) | Snp marker and application thereof | |
| WO2023001209A1 (en) | Gene chip, molecular probe combination, test kit and application for analyzing sheep fat tails | |
| WO2023070857A1 (en) | Molecular marker for distinguishing guizhou black goat from lezhi black goat, and detection method therefor and use thereof | |
| CN108410996A (en) | With the relevant InDel molecular labelings of goat production suede character and its application | |
| CN113502335A (en) | Molecular marker related to sheep growth traits and application thereof | |
| US11542562B2 (en) | Single nucleotide polymorphism marker related to Chinese horse short stature trait and use thereof | |
| CN111662966A (en) | Construction method and application of wheat backbone parent functional gene fingerprint spectrum | |
| CN110129456A (en) | A kind of anti-vibrios molecular labeling combination of prawn and its application in breeding | |
| KR101979218B1 (en) | Composition for determining bud mutation cultivar of Fuji apple | |
| CN114150070A (en) | SNP molecular marker related to chicken growth and slaughter traits, detection primer, kit and breeding method | |
| CN104672315B (en) | Control gene and to Cucumber Roll palpus character related SNP marker of the cucumber without tendril character | |
| CN109706231B (en) | High-throughput SNP (single nucleotide polymorphism) typing method for molecular breeding of litopenaeus vannamei | |
| CN107881256A (en) | For identifying single nucleotide polymorphism site, primer pair, kit and the application of Peach fruits hardship benevolence/sweet tea benevolence character | |
| KR20190045957A (en) | Single nucleotide polymorphism markers associated with daily weight gain trait in pig and use thereof | |
| CN107142326A (en) | One kind Du pool sheep SNP marker and its screening technique and application | |
| Hassan et al. | Genetic diversity among some horse breeds in Pakistan | |
| CN105483281B (en) | It is a kind of to be used to identify glutinous No. 1 SNP marker of five firework of waxy corn Shanghai and its identification method | |
| CN108841983A (en) | A kind of SSR primer of sugarcane overall length transcript profile data large-scale development |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |