CN109880915A - One kind SNP site relevant to the high character of donkey body and its application - Google Patents
One kind SNP site relevant to the high character of donkey body and its application Download PDFInfo
- Publication number
- CN109880915A CN109880915A CN201910263880.4A CN201910263880A CN109880915A CN 109880915 A CN109880915 A CN 109880915A CN 201910263880 A CN201910263880 A CN 201910263880A CN 109880915 A CN109880915 A CN 109880915A
- Authority
- CN
- China
- Prior art keywords
- donkey
- snp site
- snp
- genotype
- 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.)
- Withdrawn
Links
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses a kind of SNP sites relevant to the high character of donkey body, the SNP site is located at No. 8 chromosome of donkey, position is the 42726257th of nucleic acid, and the SNP site is base A or G, and the AA genotype individuals body height of the SNP site is significantly higher than the individual that genotype is AG and GG.The invention also discloses the primer pairs for detecting the SNP site, kit.Primer pair, kit using the SNP site and for detecting the SNP site can identify the body Gao Xingzhuan of donkey, promote breeding process.Molecular marker assisted selection of the detection of the SNP site also for the high character of donkey body provides scientific basis simultaneously.
Description
Technical field
The present invention designs genetic engineering and molecular biology field, and in particular to a kind of SNP relevant to the high character of donkey body
Site and its application.
Background technique
Family donkey (Equus asinus) belongs to Mammalia, Perissodactyla, equine, Equus, and image is like horse, but the big ear length of head, chest
Portion is slightly narrow, and trunk is shorter, body is high and body length is generally equalized.Donkey have important economic value, donkey hide be production rare Chinese medicine " Ah
The important source material of glue "；Donkey meat contains a high proportion of unsaturated fatty acid, and nutritive value is high, and Fresh ear field content is high, taste
It is delicious.There is the donkey local varieties genetic resources of very abundant in China, and Animal Genetic Resources in China will (horse donkey camel will) records me
State-owned 24 local donkey kinds.Man, China donkey is distributed in north temperate zone drying, warm region, from the east of Bohai Sea Gulf, west to Tarim Basin basin
Around ground, North gets the west of Liaoning, Ji Bei, Yan Bei, the river bend, south to the southern regions of the Yunnan Province.Large-scale donkey kind is mainly distributed on Middle-lower Reaches of Yellow River, day
Mountain southern foot and southern Tarim Basin also have the concentration place of production.Small-sized donkey north and south various regions in distributed area.Medium-sized donkey producing region all exists
Large-scale donkey producing region is neighbouring.This Distribution Pattern is limited by various regions ecological condition and economy, life background.
China's donkey variety source is abundant, but China's donkey legacy improved plan not system, each kind do not carry out systematization,
The cultivation of specialization produces meat, production skin performance does not protrude.Cultivating and producing the superior donkey new varieties of skin meat production is to ensure China donkey
The important measure that industry develops in a healthy way.Research shows that the body height of donkey and the correlation of bust, body length are very high, related coefficient is reachable
0.9 or more, bust and body are long (donkey hide area ≈ bust × body is long) directly proportional to donkey hide area；The phase relation of donkey body height and weight
Number is also 0.9 or more.It is big to cultivate physique, it is nervous can to alleviate donkey hide supply for the high specialized donkey kind of skin rate, dressing percentage out
Status, and can increase donkey meat yield, cultivate the important directions that large-scale donkey is China's donkey industry development.
Domestic scholars have also carried out various researchs to the genetic breeding of horse, and identify the time for influencing Ma Tigao character
Select gene, such as Shox, Hmga2, Tbx3, Lcorl.The high phenotype of Debao pony body especially can be explained in Hmga2 and TBX3 to become
Different 83.3%.But the research about the high trait molecular marker of donkey body is rarely reported.
Traditional selection and use is the effective means for carrying out genetic improvement to animals and plants, cultivating new varieties.In recent years, with section
The development of technology, molecular mark are gradually applied on the genetic improvement of animals and plants.Molecular mark
Have many advantages, such as that the period is short, selection traits stablize heredity.The Successful Practice of molecular mark is attributed to the fact that quantitative character number
Learn model, molecular markers for identification technology, determined dna sequence technology, micro-array chip manufacturing technology and biological information theory
Form the fast development with cross disciplines such as software developments.The precondition of molecular mark practice is positioning and character
Associated molecular labeling and the genetic mechanism for parsing character.The association analysis of candidate genes polymorphism site, quantitative trait locus
(quantitative trait loci, QTL) positioning and whole-genome association (genome-wide association
Scan, GWAS) it is widely used in the genetic mechanism of interpretive model species and non-mode species qualitative character and quantitative character, and
Achieve quite plentiful and substantial research achievement.
Summary of the invention
It is an object of the present invention to provide a kind of SNP site relevant to the high character of donkey body and its applications.
It is another object of the present invention to provide the primers for detecting the SNP site relevant to the high character of donkey body
Pair and the detection kit containing the primer pair.
To achieve the goals above, the technical solution of the present invention is as follows:
The relevant SNP site of the one high character of breeding ass body, which is characterized in that the SNP site is base A or G, the SNP
Site is located at No. 8 chromosome of donkey, and position is the 42726257th of nucleic acid；The AA genotype individuals body of the SNP site is high
It is significantly higher than the individual that genotype is AG and GG.
The present invention also provides the primer pair for detecting the SNP site relevant to the high character of donkey body, the primer packets
It includes:
Forward primer F:5 '-CGGGTAAGGCAGATCCTGAA-3 '；
Reverse primer R:5 '-TACATGGAAGGGGGAAGCAC-3 '.
The present invention also provides the detection kits for containing above-mentioned primer pair.
The present invention also provides above-mentioned SNP site, above-mentioned primer pair or detection kits containing above-mentioned primer pair to reflect
Application in the high character of donkey body.The application includes the following steps:
(1) genomic DNA of donkey to be measured is extracted；
(2) using the genomic DNA of donkey to be measured as template, using the primer pair F and R or contain the primer pair F's and R
Detection kit amplifies the chromosome 42726105 of donkey No. 8 to 214bp piece between 42726318 bit bases by PCR reaction
Section；
(3) pcr amplification product is detected, if the genotype in amplified production sequence at 153bp is AA, donkey to be measured is body
High advantage individual.
Wherein, the reaction system of PCR amplification is 20 μ L systems in the step (2)；DNA profiling 1.0 μ L, PCR
DdH is added in 10 μ L, F primer of Mixture, each 0.5 μ L of R primer2O to total volume be 20 μ L.
The reaction condition of PCR amplification in the step (2) are as follows: 95 DEG C of 5min；95 DEG C of 30s, 59 DEG C of 30s, 72 DEG C of 1min, 35
A circulation；72℃5min；4 DEG C of preservations.
Detection pcr amplification product is detected using Sanger PCR sequencing PCR in the step (3), determines to be located at donkey No. 8 dyeing
It is AG and the body of the donkey of GG height that the body height for the donkey that the 42726257th genotype is AA on body, which is significantly higher than genotype,.
The present invention further provides above-mentioned SNP site, above-mentioned primer pair or detection kits containing above-mentioned primer pair to exist
Application in donkey molecular mark.
The SNP site that the present invention is using No. 8 chromosome the 42726257th of donkey carries out Genotyping, and to the site with
The high character of donkey body is associated analysis, significance test is carried out by body height of the R lingware to different genotype individual, through aobvious
Work property check analysis discovery AA genotype individuals body height is significantly higher than AG and GG genotype individuals (P < 0.05).The SNP site is
The donkey for cultivating Gao Tigao provides important molecular labeling, will accelerate the genetic improvement process of donkey.
The invention has the benefit that the present invention provides a kind of SNP marker relevant to the high character of donkey body and its application,
SNP site relevant to the high character of donkey body is screened using polymerase chain reaction (PCR) sequencing technologies, and passes through Sanger
Sequencing technologies detect the genotype of donkey to be measured, according to genotype carry out breeding Gao Tigao donkey kind, accelerate its genetic improvement into
Journey.
Detailed description of the invention
Fig. 1 is 6 donkey individual PCR product electrophoresis detection results, wherein a1, a2 are AA genotype, and a3, a4 are AG gene
Type, a5, a6 are GG genotype；
Fig. 2 is the sequencing result of three kinds of genotype (AA, AG, GG) in PCR product.
Specific embodiment
Technical solution of the present invention is clearly and completely described below in conjunction with specific embodiment, it is clear that described
Embodiment be only a part of the embodiment of the present invention, instead of all the embodiments.Based on the embodiment of the present invention, this field
Those of ordinary skill's every other embodiment obtained, belongs to protection scope of the present invention.
If technological means used is conventional hand well known to those skilled in the art without specified otherwise in following case study on implementation
Section.
If without specified otherwise in the following example, material used and reagent etc. are commercially bought.
Embodiment 1: the Screening analysis of SNP site relevant to the high character of donkey body and its breeding side for assisting Gao Tigao donkey
Method, including the following steps:
(1) donkey extracting genome DNA；
(2) SNP site chr8:g.42726257A > G screening；
(3) design of primers；
(4) SNP site chr8:g.42726257A > G parting；
(5) correlation analysis of SNP site chr8:g.42726257A > G genotype and the high character of body；
(6) SNP site chr8:g.42726257A > G assists the selection of Gao Tigao donkey.
Concrete operations are as follows:
(1) extraction of donkey genomic DNA
(a) the musculature about 0.1g of donkey is taken, 500 μ l STE lysis buffers are added, shreds, sequentially adds 50 μ l
10%SDS, 5 μ l Proteinase Ks (20mg/ml), 56 DEG C of cracking about 3h, until lysate is clarified；
(b) same volume saturated phenol (250 μ l) being added, chloroform/isoamyl alcohol (24:1) (250 μ l) shakes gently 20min,
12000rpm is centrifuged 10min.Supernatant is taken, is repeated the above steps, until without protein layer between water phase and organic phase；
(c) supernatant is taken, same volume chloroform/isoamyl alcohol is added, gently shakes 20min, 12000rpm is centrifuged 10min；
(d) take supernatant, 1/10 volume 3M NaAc (pH5.2) and 2 times of cold dehydrated alcohols of volume be added, shake up afterwards -20 DEG C it is quiet
20min is set, 12500rpm is centrifuged 20min；
(e) nucleic acid is deposited in tube bottom, abandons supernatant, precipitated with 70% ethanol washing；
(f) precipitating is collected, air drying to ethyl alcohol all volatilizees；
(g) 20 μ l TE (A containing RNase) dissolving DNAs are added, after 37 DEG C of standing about 30min, 4 DEG C are saved；
(h) DNA sample is detected with 1% agarose gel electrophoresis, with UV spectrophotometer measuring concentration and purity.
(2) SNP site screening
The chromosome 42726105 of donkey No. 8 to 42726318 bit sequences and donkey are resurveyed sequence snp database to carry out
Blastn is compared, and screening obtains candidate SNP locus, one of them is site chr8:g.42726257A > G.
(3) design of site chr8:g.42726257A > G parting primer
According to the gene order near SNP site chr8:g.42726257A > G, serotype specific primer is designed.Standard is as follows: drawing
Object length 19-26bp；Candidate SNP locus and deletion segment are not included in primer sequence；Amplified production length 70-250bp；GC contains
Measure 30%-70%.Serotype specific primer are as follows:
Forward primer F:5 '-TACTGTAGATGGGTTCACCGATCCC-3 '；
Reverse primer R:5 '-TACATGGAAGGGGGAAGCAC-3 '.
(4) SNP site chr8:g.42726257A > G parting
With large-scale donkey 26, the genomic DNA of small-sized donkey 23 is template, utilizes the primers F and R designed in step (3)
PCR amplification is carried out, reaction system is as follows:
Add H2O complements to 20 μ l.
Amplified reaction is completed in Applied Biosystem PCR system, and reaction condition is as follows: 95 DEG C of 5min；95℃
30s, 59 DEG C of 30s, 72 DEG C of 1min, 35 circulations；72℃ 5min.
The genotype of measurement pcr amplification product is sequenced by Sanger.
(5) correlation analysis of SNP site chr8:g.42726257A > G genotype and the high character of body
25 large-scale donkeys and 25 small-sized donkeys are selected, are examined using One-way ANOVA, site of analysis chr8:
G.42726257A > G different genotype mesosome High Defferential examines chr8:g.42726257A > G related to the high character of donkey body
Property, final to determine in the individual that site chr8:g.42726257A > G genotype is AA, the high character value of body is all remarkably higher than AG
(1) P < 0.05, is shown in Table for type and GG type individual.
The body high level of the donkey of the site table 1.chr8:g.42726257A > G different genotype
Note: body height is indicated with its mean+SD in table, when shoulder is designated as different lowercases, indicates significant difference (P
< 0.05).
(6) chr8:g.42726257A > G assists the selection of Gao Tigao donkey
During the selection and use of Gao Tigao donkey, chr8:g.42726257A > in site is carried out to donkey breeding candidate population
G parting, it is preferential to select site chr8:g.42726257A > G equal in conjunction with the other and high character related locus of body parting information
It avoids selecting the site being the individual of AG type and GG type as parent as the parent of Gao Tigao donkey breeding for the individual of AA type.
SNP primer of the invention has been used for the breeding of Gao Tigao donkey.
The foregoing is merely illustrative of the preferred embodiments of the present invention, is not intended to limit the invention, all in essence of the invention
Within mind and principle, all any modification, equivalent substitution, improvement and etc. be should all be included in the protection scope of the present invention.
Claims (6)
1. the relevant SNP site of a high character of breeding ass body, which is characterized in that the SNP site be base A or G, it is SNP described
Point is located at No. 8 chromosome of donkey, and position is the 42726257th of nucleic acid.
2. SNP site as described in claim 1, which is characterized in that the AA genotype individuals body height of the SNP site is significantly high
In the individual that genotype is AG and GG.
3. a kind of primer pair for SNP site described in detecting as claimed in claim 1 or 22, which is characterized in that the primer includes:
Forward primer F:5 '-CGGGTAAGGCAGATCCTGAA-3 '；
Reverse primer R:5 '-TACATGGAAGGGGGAAGCAC-3 '.
4. a kind of kit for SNP site described in detecting as claimed in claim 1 or 22, which is characterized in that the kit includes
Primer pair as claimed in claim 3.
5. SNP site of any of claims 1 or 2, primer pair as claimed in claim 3 or kit as claimed in claim 4,
Application in the identification high character of donkey body.
6. SNP site of any of claims 1 or 2, primer pair as claimed in claim 3 or kit as claimed in claim 4,
Application in donkey molecular mark.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910263880.4A CN109880915A (en) | 2019-04-01 | 2019-04-01 | One kind SNP site relevant to the high character of donkey body and its application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910263880.4A CN109880915A (en) | 2019-04-01 | 2019-04-01 | One kind SNP site relevant to the high character of donkey body and its application |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN109880915A true CN109880915A (en) | 2019-06-14 |
Family
ID=66935902
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910263880.4A Withdrawn CN109880915A (en) | 2019-04-01 | 2019-04-01 | One kind SNP site relevant to the high character of donkey body and its application |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109880915A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112284501A (en) * | 2020-09-25 | 2021-01-29 | 聊城大学 | System and method for predicting body weight of donkey |
| CN114934122A (en) * | 2022-06-23 | 2022-08-23 | 新疆农业大学 | Kit containing ADCY8 gene for screening lactation traits of Xinjiang donkey and application of kit |
-
2019
- 2019-04-01 CN CN201910263880.4A patent/CN109880915A/en not_active Withdrawn
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112284501A (en) * | 2020-09-25 | 2021-01-29 | 聊城大学 | System and method for predicting body weight of donkey |
| CN114934122A (en) * | 2022-06-23 | 2022-08-23 | 新疆农业大学 | Kit containing ADCY8 gene for screening lactation traits of Xinjiang donkey and application of kit |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107326077A (en) | A kind of molecular labeling for differentiating spotted maigre genetic sex and its application | |
| CN107227373B (en) | SNP functional molecular marker of japonica rice lodging-resistant gene and application | |
| CN105802960B (en) | Molecular labeling and its application | |
| CN112609009B (en) | SNP molecular marker combination related to weight and body size of Guangxi three-yellow chicken based on whole genome sequencing screening and application | |
| CN104561355B (en) | Multiplex-PCR method for parentage assignment of scapharca broughtonii | |
| CN105821054B (en) | sinkiang bream DNA bar code standard detection sequence and application thereof | |
| CN109554486A (en) | SNP marker relevant to grass carp character and its application | |
| CN114182032A (en) | SNP molecular marker for detecting seed coat color of muskmelon and application thereof | |
| CN105802962B (en) | Molecular labeling and its application | |
| CN106498048B (en) | One kind QTL relevant to soybean nodulation number, SNP marker and application | |
| CN104651356B (en) | A kind of SNP marker related to Erhualian sow litter trait and its detection method and application | |
| CN109880915A (en) | One kind SNP site relevant to the high character of donkey body and its application | |
| CN104672315B (en) | Control gene and to Cucumber Roll palpus character related SNP marker of the cucumber without tendril character | |
| CN115011717A (en) | Development and detection method and breeding application of watermelon whole-genome SNP-Panel | |
| CN110747282A (en) | Low-salt-resistant molecular marker C22 of portunus trituberculatus and application thereof | |
| CN104846082B (en) | A kind of Dermestidae insect genes barcode detection kit | |
| CN105802961B (en) | Molecular labeling and its application | |
| CN116200528B (en) | SNP molecular marker linked with wheat stripe rust resistance gene QYr.sicau. -2BL and application thereof | |
| CN116769891A (en) | SNP (Single nucleotide polymorphism) marker for rapidly identifying genetic sex of Chinese soft-shelled turtles as well as primer and application thereof | |
| CN106834521A (en) | A kind of SNP marker of Odontobulis mpotamophila growth traits related gene and its amplimer and application | |
| 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 | |
| CN103695416B (en) | A kind of method and its application for the SNP for detecting Qinchuan Cattle CFL2 genes | |
| CN104694651B (en) | A kind of SNP marker related to Erhualian sow litter trait, detection method and application | |
| CN112210607B (en) | Molecular marker related to buffalo white hair phenotype and application thereof | |
| CN108546778B (en) | SNP molecular marker for detecting powdery mildew resistance of cucumber and application thereof |
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 | ||
| WW01 | Invention patent application withdrawn after publication |
Application publication date: 20190614 |
|
| WW01 | Invention patent application withdrawn after publication |