CN106498067B - The method for cultivating capital sea yellow chicken low fat system using THRSP α gene - Google Patents
The method for cultivating capital sea yellow chicken low fat system using THRSP α gene Download PDFInfo
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Abstract
The present invention relates to field of poultry breeding, and in particular to a method of capital sea yellow chicken low fat system is cultivated using THRSP α gene.Application the invention discloses THRSP α gene extron 2 as the yellow chicken abdominal fat weight in capital sea and the molecular genetic marker of abdominal fat.And capital sea yellow chicken low fat system is cultivated using the molecular labeling; it is respectively the homozygous individual of T and G allele at 129 and 160bp of selection THRSP α gene extron 2; the genotype can be fixed in offspring; the method of the present invention significant effect; it is simple and fast; and it is free from the influence of the external environment, applied to the quality broiler chicken strain for cultivating low abdominal fat.
Description
Technical field
The invention belongs to field of poultry breeding, and in particular to a kind of to cultivate capital sea yellow chicken low fat system using THRSP α gene
Method.
Background technique
Past, most of breeding experts were primarily upon the speed of growth of chicken and the increase situation of weight, but had ignored chicken fat
The selection of character especially stomach fat character, therefore while the chicken speed of growth is constantly accelerated, also produce fat deposition
Excessive problem.Fat deposition occurs mainly in the positions such as the subcutaneous of chicken, internal organ, muscle and bone, wherein chicken subcutaneous abdomen
It is the main portions of fat deposition.Abdominal Fat in Broilers deposition excessively can not only reduce efficiency of feed utilization, but also have an effect on Tu of broiler chicken
Body character, laying hen overfertilization will affect the Age at first laying and egg production of laying hen, therefore increasingly to the genetic improvement of chicken ventral fat character
Attention by breeding scholar and breeding enterprise.Although conventional breeding methods obtain certain progress in poultry genetic improvement,
But this method breeding cycle is long, costly, and molecular marker-assisted selection method not only reduces breeding cost, can more accelerate
Breeding process provides strong tool for poultry breeding work.
Thyroid hormone response protein Spot 14 (THRSP) is that a participation regulation fat generates enzymatic activity and then influences
The transcription regulatory factor of Tissue Lipids metabolism, THRSP α are a kind of hypotypes of Spot 14, and the albumen of THRSP α gene coding is one
Kind small-molecular-weight, acid cell nuclear protein, it is just regulating and controlling or the auxiliary activating transcription factor of negative regulation, in tissue specificity lipid
Crucial effect is played in metabolic regulation.
Summary of the invention
The technical problem to be solved in the present invention is to provide a kind of sides that capital sea yellow chicken low fat system is cultivated using THRSP α gene
Method.This method utilizes the different genotype for being mutated and generating at the C129T and T160G two occurred on THRSP α gene extron 2,
Assisted Selection is marked to the abdominal fat weight and abdominal fat of chicken, this method is not affected by environment.
Molecular genetic marker the invention discloses THRSP α gene extron 2 as capital sea yellow chicken abdominal fat weight and abdominal fat
Application.The application is specifically the 129 and 160bp that THRSP α gene extron 2 is selected when cultivating capital sea Huang chicken low fat system
Place is respectively the homozygous individual of T and G allele.
The invention discloses mutational site at C129T and T160G two on THRSP α gene extron 2 as with the yellow chicken in capital sea
The application of abdominal fat weight and the significant relevant specific molecular marker of abdominal fat.
It is that selection capital sea is yellow the invention also discloses a kind of method for cultivating capital sea yellow chicken low fat system using THRSP α gene
In chicken at 129 and 160bp of THRSP α gene extron 2 be respectively T and G allele homozygous individual based on group, warp
After crossing the seed selection and breeding of multiple generations, until low ventral fat character is fixed, each generation is utilized in this Breeding Process
It is respectively that T and G is selected at 129 and 160bp of THRSP α gene extron 2.
It is as follows to the molecular mark detection method of THRSP α gene extron 2: the chicken genomic DNA of sample to be tested is extracted,
The target fragment of 221bp is obtained through sequence primer amplified as shown in SEQ ID NO:1 and SEQ ID NO:2；Purpose
Segment is developed the color after gel electrophoresis with silver staining through sscp analysis, obtains the SSCP map of DNA, screens base according to the banding pattern in map
Because of the individual that type is DD；Either direct Sequencing is respectively T and G equipotential at 129 and 160bp of THRSP α gene extron 2
The individual of gene.
The principle of the present invention is: pair of primers is devised for THRSP α gene extron 2, with the yellow chicken full-length genome in capital sea
DNA is that template is expanded, and amplified production generates 6 kinds of genotype (AA, BB, CC, DD, AB and CD) through sscp analysis, directly surveys
Sequence shows that C → T mutation has occurred in BB type compared with AA type at 129bp, and it is prominent that T → G occurs at 160bp compared with AA type for CC type
Become, DD type occurs to be mutated at C → T and T → G two respectively at 129bp and 160bp compared with AA type.By different genotype and capital
Extra large Huang chicken abdominal fat weight and abdominal fat carry out correlation analysis (since BB genotype individuals number is less than 3, being not involved in correlation analysis), analysis
The result shows that DD genotype individuals abdominal fat weight and abdominal fat are minimum, and extremely significant lower than other genotype individuals (P < 0.01).It should
The significant effect of selection, detection method is simple and fast, and free from the influence of the external environment, can be used for detecting the low abdomen of the yellow chicken in capital sea
The molecular genetic marker of rouge rate.
Detailed description of the invention
Fig. 1 is the SSCP map of THRSP α gene.
Fig. 2 is the sequencer map of THRSP α gene.
Specific embodiment
Embodiment 1
1. test material
Raising is chosen in Jiangsu Jinghai Poultry Group Co., Ltd. with a batch of yellow chicken in 379 capital sea, carries out wing venous
Blood sampling and slaughter determining.Record and arrange the Carcass Traits index of this batch of chicken.Chicken genome is extracted using phenol chloroform method
DNA, is dissolved in TE, and NANODROP1000 nucleic acid concentration analyzer measures -20 DEG C of preservations after concentration and purity.
2. design of primers and PCR amplification
The design of 2.1 SSCP primers
1 pair is designed according to 2 sequence of THRSP α gene extron (accession number: NC_006088.3) that GenBank has been delivered
Primer, amplified production clip size are 221bp.Primer sequence is as follows:
F:5 '-ACTGTGTCCGTTCTCCCAAC-3 ' (SEQ ID NO:1)
R:5 '-CTCATTTCTGCCCTGCCTAC-3 ' (SEQ ID NO:2)
1.2 PCR amplification
Pcr amplification reaction system is 20 μ L:10 × buffer (25mmol/L), 2 μ L；Mg2+(10pmol/μL)2.2μL；
0.8 μ L of dNTPs (2.5mmol/L), 1 μ L of DNA profiling, primer (being 10pmol/L) each 1 μ L, Taq DNA polymerase (5U/ μ L)
0.2 μ L, 11.8 μ L of distilled water.Amplification program: 94 DEG C of initial denaturation 5min；94 DEG C of denaturation 30s, anneal 30s, 72 DEG C of extension 30s, and 30
A circulation；Last 72 DEG C of extensions 10min, 4 DEG C of preservation products.Primer annealing temperature is 60 DEG C.Polyacrylamide gel electrophoresis inspection
Survey amplified production.
2.3 PCR-SSCP detection
2.5 μ L PCR products and 7.5 μ L sample-loading buffers are mixed, after 98C is denaturalized 10min, rapid ice bath 10min.It takes
7 μ L of PCR product after denaturation carries out native polyacrylamide gel electrophoresis, first uses 250V voltage prerunning 5min, then use
After 10~12h of electrophoresis of 110V, silver staining colour developing.
The SSCP map of primer amplification segment as shown in Figure 1,6 kinds of genotype of display, be respectively defined as AA, BB, CC, DD,
(banding pattern suggests the formation of 6 kinds of genotype to AB and CD genotype, and four kinds homozygous to be indicated with AA, BB, CC and DD respectively, and two kinds miscellaneous
Mould assembly indicates that allele of the AA type at 129 and 160bp is respectively C and T with AB and CD, and BB type is distinguished in corresponding site
For T and T, CC type is respectively C and G in corresponding site, and DD type is respectively T and G in corresponding site).Each genotype PCR is produced
Object segment is sequenced, as a result, it has been found that: BB detects that C129T is mutated compared with AA in exon 2, and CC is shown outside compared with AA
Detect that T160G is mutated in son 2, DD detects mutation (Fig. 2) at C129T and T160G two compared with AA in exon 2.
The yellow chicken abdominal fat weight of 2.4 each genotype of THRSP α gene and capital sea and abdominal fat association analysis
With following model: Yij=μ+Gj+eij, something lost of the analysis each genotype of THRSP α gene to the yellow chicken ventral fat character in capital sea
Pass effect, wherein YijFor the record value of ventral fat character；μ is community average；GiFor genotype effects, eijIt is random error.Capital
Extra large Huang chicken DD genotype is shown in Table 1 compared with other genotype abdominal fat weights and abdominal fat.
The comparison of 1 capital of table sea yellow chicken THRSP α gene different genotype abdominal fat weight and abdominal fat
Note: colleague's data shoulder, which infuses different capitalization persons, indicates that difference is extremely significant (P < 0.01), and shoulder infuses different lowercases
Person indicates significant difference (P < 0.05).
In all genotype, the abdominal fat of DD genotype weight and abdominal fat be it is minimum, the abdominal fat of DD genotype compares AB again
The low 41.89g of genotype (P < 0.01), low 3.59% (P < 0.01) of abdominal fat ratio AB genotype, DD genotype abdominal fat than 5 kinds bases again
Because type is averaged the abdominal fat weight low 4.92g of 24.54g, than 5 kinds genotypic mean abdominal fats of abdominal fat 4.52% low 2.32%.Screening
THRSP α gene DD genotype can reduce the yellow chicken abdominal fat weight in capital sea and abdominal fat, so being schemed according to the SSCP of chicken THRSP α gene
Spectrum screening DD genotype can be used as the method for cultivating capital sea yellow chicken low fat system.
SEQUENCE LISTING
<110>Yangzhou University
<120>method for cultivating capital sea yellow chicken low fat system using THRSP α gene
<130>
<160> 2
<170> PatentIn version 3.3
<210> 1
<211> 20
<212> DNA
<213>artificial sequence
<400> 1
actgtgtccg ttctcccaac 20
<210> 2
<211> 20
<212> DNA
<213>artificial sequence
<400> 2
ctcatttctg ccctgcctac 20
Claims (2)
- Application of the 1.THRSP α gene extron 2 as the yellow chicken abdominal fat weight in capital sea and the molecular genetic marker of abdominal fat, feature Be, in the application select THRSP α gene extron 2 the 129th and 160bp place respectively T and G allele homozygosis It according to accession number in GenBank is NC_ 006088.3 at individual, the THRSP α gene extron 2 the 129th and 160bp THRSP α gene order and determine.
- 2. a kind of method for cultivating the yellow chicken low fat system in capital sea using THRSP α gene, which is characterized in that in the yellow chicken in selection capital sea Be respectively at THRSP α gene extron 2 the 129th and 160bp T and G allele homozygous individual based on group, process is multiple After the seed selection and breeding of generation, until low ventral fat character is fixed, each generation utilizes THRSP α base in this Breeding Process Because being respectively that T and G is selected at exon 2 the 129th and 160bp, at the THRSP α gene extron 2 the 129th and 160bp It is determined according to the THRSP α gene order that accession number in GenBank is NC_ 006088.3.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101921770A (en) * | 2010-04-27 | 2010-12-22 | 曲湘勇 | PPAR (Peroxisome Proliferator-Activated Receptor) alpha gene and application thereof as goose fat traits genetic markers |
| CN102106296A (en) * | 2011-03-01 | 2011-06-29 | 安徽农业大学 | Method for cultivating novel Dingyuan pig breed |
| CN103146829A (en) * | 2013-03-11 | 2013-06-12 | 浙江省农业科学院 | Method for assistant selection of goose muscle fatty acid performances by utilizing molecular markers |
-
2016
- 2016-11-10 CN CN201610990134.1A patent/CN106498067B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101921770A (en) * | 2010-04-27 | 2010-12-22 | 曲湘勇 | PPAR (Peroxisome Proliferator-Activated Receptor) alpha gene and application thereof as goose fat traits genetic markers |
| CN102106296A (en) * | 2011-03-01 | 2011-06-29 | 安徽农业大学 | Method for cultivating novel Dingyuan pig breed |
| CN103146829A (en) * | 2013-03-11 | 2013-06-12 | 浙江省农业科学院 | Method for assistant selection of goose muscle fatty acid performances by utilizing molecular markers |
Non-Patent Citations (3)
| Title |
|---|
| Duplicated Spot 14 genes in the chicken- characterization and identification of polymorphisms associated with abdominal fat traits;Xiaofei Wang等;《Gene》;20040401;第332卷;79-88 * |
| 鸡、鸭甲状腺激素应答基因(THRSP)的研究进展;詹凯等;《遗传》;20090228;第31卷(第2期);131-136 * |
| 鸡THRSP α基因多态性与脂肪性状的相关性研究;闫文龙等;《西南大学学报（自然科学版）》;20070228;第29卷(第2期);57-60 * |
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