CN104480108B - Method for identifying pig back fat thickness and special primer pair thereof - Google Patents
Method for identifying pig back fat thickness and special primer pair thereof Download PDFInfo
- Publication number
- CN104480108B CN104480108B CN201410769353.8A CN201410769353A CN104480108B CN 104480108 B CN104480108 B CN 104480108B CN 201410769353 A CN201410769353 A CN 201410769353A CN 104480108 B CN104480108 B CN 104480108B
- Authority
- CN
- China
- Prior art keywords
- pig
- genotype
- homozygosis
- thickness
- base
- 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.)
- Expired - Fee Related
Links
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses a method for identifying the pig back fat thickness and a special primer pair thereof. The invention discloses a primer pair for amplifying a DNA fragment with g.13739C>T polymorphic site, wherein the g.13739C>T polymorphic site is the 13739th site from the 5' end of a sequence with GenBankAccession Number NC_010449.4; and the GenBankAccession Number is the update time (September 29, 2013) of the sequence NC_010449.4. By adopting the method disclosed by the invention to breed pigs, the candidate pigs can be screened early, so that the breeding cost is reduced, and the back fat thickness of the pigs in actual production can be effectively reduced or increased. The method disclosed by the invention is simple to operate, requires low cost, has high accuracy, can realize automatic direct detection, and plays a great role in pig breeding.
Description
Technical field
The present invention relates to a kind of method of identification fat thickness at back of pig size and its special primer pair, belong to biological technical field.
Background technology
The thickness of backfat of pig is always the important component part of pig breeding target, and the thickness of back fat is not only relevant with lean meat percentage,
Impact for the speed of growth is also very huge, and therefore its Breeding value is very high.But, because the indexs such as the thickness of backfat can only be in animal
Could measure after growing up to, on the one hand increase breeding cost, on the other hand elongate the generation inteval, genetic progress is slow.Point
The structure developing rapidly with pig genetic linkage mapses of sub- biotechnology, be the quantitative characters such as the thickness of backfat carry out breeding more accurately,
Faster molecular breeding provides Research foundation to progress.On the other hand, because pig is a kind of important pattern of human research
Animal, therefore, finding pig gene influential on back fat and being possible to the research of the obesity to the mankind provides certain reference.
Content of the invention
It is an object of the invention to provide a kind of method of identification fat thickness at back of pig size and its special primer pair.
The present invention provides a kind of amplification containing g.13739c > primer pair of the dna fragment of t pleomorphism site；
Described g.13739c > t pleomorphism site be sequence 3 or 4 in the 214th from 5 ' ends.
In above-mentioned primer pair, shown in dna molecule shown in seq id no.1 for the described primer pair and seq id no.2
Dna molecular composition.
A kind of kit of the thickness of backfat size of identification or auxiliary identification pig falls within protection scope of the present invention, this reagent
Box comprises any of the above-described described primer pair.
A kind of method of the thickness of backfat size of identification or auxiliary identification pig falls within protection scope of the present invention, the method
For: include detecting the genotype of pig to be measured, determine the back fat thickness of pig, homozygosis tt gene according to the genotype of described pig to be measured
The thickness of backfat of the pig of type is more than the thickness of backfat of the pig of heterozygosis ct genotype, and the thickness of backfat of the pig of heterozygosis ct genotype is more than homozygosis cc
The thickness of backfat of the pig of genotype；
G.13739c the pig of described homozygosis cc genotype is > base of t pleomorphism site is the pig of c；
G.13739c the pig of described heterozygosis ct genotype is > base of t pleomorphism site is the pig of t and c；
G.13739c the pig of described homozygosis tt genotype is > base of t pleomorphism site is the pig of t；
Described g.13739c > t pleomorphism site be sequence 3 or 4 in the 214th from 5 ' ends.
In said method, the decision method of described homozygosis cc genotype, heterozygosis ct genotype or homozygosis tt genotype is as follows:
With the genome dna of pig as template, pcr amplification is carried out for primer with above-mentioned primer, obtain pcr amplified production, if pcr amplification
Product base of the 214th from 5 ' ends is c, then the genotype of described pig is homozygosis cc genotype, if pcr amplification is produced
Thing base of the 214th from 5 ' ends is t and c, then the genotype of described pig is heterozygosis ct genotype, if pcr amplification is produced
Thing base of the 214th from 5 ' ends is t, then the genotype of described pig is homozygosis tt genotype.
A kind of method of the little pig of seed selection thickness of backfat falls within protection scope of the present invention, is to select homozygosis cc genotype
Pig carries out seed selection；
G.13739c the pig of described homozygosis cc genotype is > base of t pleomorphism site is the pig of c；
Described g.13739c > t pleomorphism site be sequence 3 or 4 in the 214th from 5 ' ends.
In said method, the decision method of described homozygosis cc genotype is as follows: with the genome dna of pig as template, above
State primer and carry out pcr amplification for primer, obtain pcr amplified production, if pcr amplified production alkali of the 214th from 5 ' ends
Base is c, then the genotype of described pig is homozygosis cc genotype.
A kind of method of the big pig of seed selection thickness of backfat falls within protection scope of the present invention, is to select homozygosis tt genotype
Pig carries out seed selection；
G.13739c the pig of described homozygosis tt genotype is > base of t pleomorphism site is the pig of t；
Described g.13739c > t pleomorphism site be sequence 3 or 4 in the 214th from 5 ' ends.
In said method, the decision method of described homozygosis tt genotype is as follows: with the genome dna of pig as template, to weigh
Profit requires the primer described in 2 to carry out pcr amplification for primer, obtains pcr amplified production, if pcr amplified production is from 5 ' ends
The base of the 214th is t, then the genotype of described pig is homozygosis tt genotype.
Any of the above-described described primer pair, mentioned reagent box and/or any of the above-described described method are in the seed selection of pig
Application falls within protection scope of the present invention.
The experiment proves that, the present invention uses gene order surveying method to detect g.13739c > t, only need to carry out pcr anti-
Should, sequencing can differentiate the genotype of individuality, and genotype sentences that type is very accurate, and testing cost is not high, has very high breeding real
Trample using value.With the method for the present invention, fat thickness at back of pig proterties is selected, the thick thinning about 6.07mm of pig average backfat can be made,
Thus obtaining the higher pig of production performance.
The method that the application present invention provides carries out breeding to pig, can carry out early screening to pig to be selected, effectively alleviates real
The problem selecting excellent boar time length in the production of border, reduces breeding and spends, the pig in effective reduction or raising actual production
Back fat trait Advances in Breeding.The detection method of the present invention is simple to operate, expense is not high, the degree of accuracy is high, and can achieve automation
Direct detection, plays a great role in the breeding work of pig.
Brief description
Fig. 1 be cc genotype individuals and tt genotype individuals nudt3 gene g.13739c > t pleomorphism site sequence nearby
Sequencing result.
Specific embodiment
Experimental technique used in following embodiments if no special instructions, is conventional method.
Material used, reagent etc. in following embodiments, if no special instructions, all commercially obtain.
Big people's hybridized pig (sus scrofa), is obtained by Large White and the hybridization of people pig, this pig is purchased from the Chinese Academy of Agricultural Sciences
Beijing animal and veterinary research institute Changping livestock and poultry Experimental Base.
Genbank accession number in following embodiments is that the sequence of nc_010449.4 each means renewal day
It is the sequence of nc_010449.4 in September, the 2013 genbank accession number of 29 days.
Embodiment 1, the identification of fat thickness at back of pig size
First, pig nudt3 gene is g.13739c > determination of t pleomorphism site
(1) with two big people hybridized pig as experiment material, extract the genome dna of its ear-edge tissue respectively.
(2) design of primer and synthesis
Believed according to pig nudt3 gene group dna sequence (genbank accession number nc_010449.4)
Breath, designs and synthesizes following primer:
U (upstream primer): 5 '-atcacttccactccactca-3 ' (seq id no.1)；
D (downstream primer): 5 '-gctctcggtctcagttct-3 ' (seq id no.2).
(3) pcr amplification
The genome dna of the big people's hybridized pig being obtained with step () respectively, as template, carries out pcr expansion with u and d for primer
Increase, obtain pcr amplified production, be respectively designated as product 1 and product 2.
Pcr amplification system: genome dna 200ng, 10 × pcr amplification buffer 5 μ l, the final concentration of 10mm of dntps,
The each 50ng of upstream and downstream primer, taq dna polymerase 0.75u, mg2+2.5mmol/l, uses ddh2O supplies system to 50 μ l.
Pcr amplification program: 95 DEG C of denaturation 5min；Then 95 DEG C of denaturation 20s, 59 DEG C of annealing 30s, 72 DEG C of extension 30s, totally 35
Individual circulation；Last 72 DEG C of extension 10min.
(4) sequencing and sequence analysis
Product 1 and product 2 are sequenced, are obtained the sequence (as shown in seq id no.3) of product 1 and the sequence of product 2
Row (as shown in seq id no.4).Seq id no.3 and seq id no.4 only exists the difference of a base, is seq id
In no.3 and seq id no.4 the 214th from 5 ' ends, the base at this is c or t, sequence such as Fig. 1 institute of this location proximate
Show (wherein arrow show this pleomorphism site).
The pcr amplified production obtaining in step (three) base of the 214th from 5 ' ends is the individuality of c, this individuality
For homozygous individuality, this individual genotype is named as homozygosis cc genotype, the pcr amplified production obtaining in g step (three)
From 5 ' ends, the base of the 214th is the individuality of t, and this individuality is homozygous individuality, and this individual genotype is named as
Homozygosis tt genotype, the pcr amplified production obtaining in step (three) base of the 214th from 5 ' ends is the individuality of t and c,
This individuality is that heterozygous is individual, and this individual genotype is named as heterozygosis ct genotype.
2nd, pig nudt3 gene is g.13739c > correlation analysis of t pleomorphism site and fat thickness at back of pig proterties
For determination g.13739c > whether t pleomorphism site is related to fat thickness at back of pig proterties, with 588 big people's hybridized pigs is
Experiment material, is tested as follows:
(1) extract the genome dna of the ear-edge tissue of every pig, carry out pcr according to the method for (three) in step one respectively
Amplification, obtains each pcr amplified production, the method according to (four) in step one determines that the genotype of every pig is homozygosis cc, heterozygosis
Ct or homozygosis tt.
(2) measure and record the thickness of backfat size of every pig.
(3) genotype according to pig and fat thickness at back of pig size obtain as drawn a conclusion: the back fat of the pig of homozygosis tt genotype
The thickness of backfat of the thick pig more than heterozygosis ct genotype, the thickness of backfat of the pig of heterozygosis ct genotype is more than the pig of homozygosis cc genotype
The thickness of backfat.
(4) to g.13739c > t site and fat thickness at back of pig proterties carry out association analysis with least square method.Model used is such as
Under:
Y=s+p+b+g+e
Wherein y is property determination value, and s is sex-effects, and p is parity effect, and b is to butcher batch effect, and g is imitated for genotype
Should, e is residual error effect.
Result is as shown in table 1.
The genotype of table 1 pig and the correlation analysis of fat thickness at back of pig
Note: same column subscript difference letter representation significant difference (p < 0.05)
Table 1 shows, in fat thickness at back of pig proterties, the fat thickness at back of pig of the pig of homozygosis tt genotype is more than heterozygosis ct genotype
The fat thickness at back of pig of pig, the fat thickness at back of pig of the pig of heterozygosis ct genotype is more than the fat thickness at back of pig of the pig of homozygosis cc genotype, homozygosis tt
The pig of genotype than homozygosis cc genotype pig in terms of fat thickness at back of pig about high about 6.07mm (p < 0.05).Illustrate that the present invention uses
Nudt3 gene (shown in sequence 3 or 4) SNP of the 214th from 5 ' ends identifies fat thickness at back of pig side
Method is consistent with the practical measurement result of fat thickness at back of pig.
In sum, in actual pig breeding, for obtaining the pig of thinner back fat thickness, it is preferably selected cc genotype
Pig carries out breeding.
Claims (9)
1. a kind of amplification is containing g.13739c > primer pair of the dna fragment of t pleomorphism site；
Described g.13739c > t pleomorphism site be sequence 3 or 4 in the 214th from 5 ' ends；
Dna molecule shown in seq id no.1 for the described primer pair and the dna molecular composition shown in seq id no.2.
2. the kit of the thickness of backfat size of a kind of identification or auxiliary identification pig, this kit comprises drawing described in claim 1
Thing pair.
3. the method for the thickness of backfat size of a kind of identification or auxiliary identification pig, the method is: includes detecting the genotype of pig to be measured,
Determine the back fat thickness of pig according to the genotype of described pig to be measured, the thickness of backfat of the pig of homozygosis tt genotype is more than heterozygosis ct base
Because of the thickness of backfat of the pig of type, the thickness of backfat of the pig of heterozygosis ct genotype is more than the thickness of backfat of the pig of homozygosis cc genotype；
G.13739c the pig of described homozygosis cc genotype is > base of t pleomorphism site is the pig of c；
G.13739c the pig of described heterozygosis ct genotype is > base of t pleomorphism site is the pig of t and c；
G.13739c the pig of described homozygosis tt genotype is > base of t pleomorphism site is the pig of t；
Described g.13739c > t pleomorphism site be listing sequence 3 or 4 in the 214th from 5 ' ends.
4. method according to claim 3 it is characterised in that: described homozygosis cc genotype, heterozygosis ct genotype or homozygosis
The decision method of tt genotype is as follows: with the genome dna of pig as template, is carried out with the primer described in claim 1 for primer
Pcr expands, and obtains pcr amplified production, if pcr amplified production base of the 214th from 5 ' ends is c, described pig
Genotype be homozygosis cc genotype, if pcr amplified production from 5 ' ends the 214th base be t and c, described pig
Genotype be heterozygosis ct genotype, if pcr amplified production base of the 214th from 5 ' ends is t, described pig
Genotype is homozygosis tt genotype.
5. the method for the little pig of a kind of seed selection thickness of backfat, is that the pig selecting homozygosis cc genotype carries out seed selection；
G.13739c the pig of described homozygosis cc genotype is > base of t pleomorphism site is the pig of c；
Described g.13739c > t pleomorphism site be listing sequence 3 or 4 in the 214th from 5 ' ends.
6. method according to claim 5 it is characterised in that: the decision method of described homozygosis cc genotype is as follows: with pig
Genome dna be template, pcr amplification is carried out for primer with the primer described in claim 1, obtains pcr amplified production, if
Pcr amplified production base of the 214th from 5 ' ends is c, then the genotype of described pig is homozygosis cc genotype.
7. the method for the big pig of a kind of seed selection thickness of backfat, is that the pig selecting homozygosis tt genotype carries out seed selection；
G.13739c the pig of described homozygosis tt genotype is > base of t pleomorphism site is the pig of t；
Described g.13739c > t pleomorphism site be listing sequence 3 or 4 in the 214th from 5 ' ends.
8. method according to claim 7 it is characterised in that: the decision method of described homozygosis tt genotype is as follows: with pig
Genome dna be template, pcr amplification is carried out for primer with the primer described in claim 1, obtains pcr amplified production, if
Pcr amplified production base of the 214th from 5 ' ends is t, then the genotype of described pig is homozygosis tt genotype.
9. the primer pair described in claim 1, the kit described in claim 2 and/or the arbitrary described side of claim 3-8
Application in the seed selection of pig for the method.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410769353.8A CN104480108B (en) | 2014-12-12 | 2014-12-12 | Method for identifying pig back fat thickness and special primer pair thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410769353.8A CN104480108B (en) | 2014-12-12 | 2014-12-12 | Method for identifying pig back fat thickness and special primer pair thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104480108A CN104480108A (en) | 2015-04-01 |
| CN104480108B true CN104480108B (en) | 2017-01-18 |
Family
ID=52754715
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410769353.8A Expired - Fee Related CN104480108B (en) | 2014-12-12 | 2014-12-12 | Method for identifying pig back fat thickness and special primer pair thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104480108B (en) |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107815499B (en) * | 2017-11-14 | 2020-05-15 | 中国农业大学 | SNP (single nucleotide polymorphism) locus related to 100kg body weight backfat thickness of pig and application thereof |
| CN107937557B (en) * | 2017-11-14 | 2020-05-05 | 中国农业大学 | SNP (single nucleotide polymorphism) site related to economic traits of pigs and application thereof |
| CN107937559B (en) * | 2017-11-14 | 2020-05-05 | 中国农业大学 | SNP (single nucleotide polymorphism) site related to economic traits of pigs and application thereof |
| CN110117667B (en) * | 2019-06-06 | 2022-04-26 | 中国农业科学院北京畜牧兽医研究所 | Method for identifying density of pig muscle fibers and primer pair used by method |
Family Cites Families (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100554420C (en) * | 2007-05-21 | 2009-10-28 | 华中农业大学 | Daily gain in pigs TEF-1 gene and the application in the pig molecule mark assisted Selection thereof |
| KR20120051232A (en) * | 2010-11-12 | 2012-05-22 | 고려대학교 산학협력단 | Dna markers for detecting decrease of androstenone within backfat in pigs |
| CN102559890B (en) * | 2011-12-28 | 2013-12-25 | 中山大学 | Method for evaluating fat deposition performance of pig |
| CN104195136B (en) * | 2014-09-04 | 2017-01-11 | 中国农业科学院北京畜牧兽医研究所 | Method or identifying length and/or height of pig body and special primer pair therefor |
-
2014
- 2014-12-12 CN CN201410769353.8A patent/CN104480108B/en not_active Expired - Fee Related
Also Published As
| Publication number | Publication date |
|---|---|
| CN104480108A (en) | 2015-04-01 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN108728552B (en) | It is a kind of influence duroc eye muscle area character molecular labeling and application | |
| CN104480108B (en) | Method for identifying pig back fat thickness and special primer pair thereof | |
| CN103146829B (en) | Method for assistant selection of goose muscle fatty acid performances by utilizing molecular markers | |
| CN102559890A (en) | Method for evaluating fat deposition performance of pig | |
| CN105087820A (en) | FSHR (follicle stimulating hormone receptor) gene based molecular marker related to porcine reproduction traits as well as detection method and application of molecular marker | |
| Wang et al. | A novel A> G polymorphism in the intron 2 of TBX3 gene is significantly associated with body size in donkeys | |
| CN108359733B (en) | Method for auxiliary detection of 100kg backfat thickness of pig | |
| CN104278027B (en) | A kind of method and its special primer pair of intramuscular fat content height of identification pig | |
| CN103421771B (en) | Genetic marker for character of litter size of pig utilizing WIF1 gene | |
| Nigussie et al. | Genetic diversity and matrilineal genetic origin of fat-rumped sheep in Ethiopia | |
| CN108588235A (en) | Highly relevant genetic marker and its application with the velour yield and pile layer of Characteristics in Ziwuling black goat | |
| CN108441566A (en) | A kind of detection method of goat ATBF1 gene insertion/deletions and its application | |
| CN110195114B (en) | SNP molecular marker influencing pig muscle fiber density and application thereof | |
| CN114921568B (en) | SNP molecular marker related to Qinchuan cattle body ruler and meat quality traits and application thereof | |
| CN104195136B (en) | Method or identifying length and/or height of pig body and special primer pair therefor | |
| CN110229914A (en) | A kind of SNP marker and its application influencing pig diameter of muscle fiber | |
| CN108220449A (en) | A kind of method and its special primer pair that detection pig is assisted to reach 100kg weight ages in days | |
| Smaragdov | Genomic selection as a possible accelerator of traditional selection | |
| CN105331696B (en) | A kind of method and primer special for identifying pig rib data/coherency shape | |
| Balteanu et al. | Characterization of the αS1-casein IRV allele provides evidence for phylogeny of the ancient Romanian Grey Steppe cattle, Moldavian strain | |
| CN110438237B (en) | SNP (single nucleotide polymorphism) site related to posttendinosus and fashion head weight on chromosome 6 of meat Simmental cattle and application | |
| CN105256045B (en) | It is a kind of identify pig kill after 24 it is small when longissimus dorsi muscle pH value size method and its special primer pair | |
| CN106434909A (en) | Molecular marker method for predicting and identifying duck growth and abdomen fat percentages and application | |
| CN114231639A (en) | Method for identifying or assisting in identifying porcine eye muscle area and application of method | |
| Dall’Olio et al. | ESR1 and ESR2 gene markers are not associated with number of piglets born alive in Italian Large White sows |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20170118 Termination date: 20181212 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |