CN106011232A - Method for selecting chicken resisting performance - Google Patents
Method for selecting chicken resisting performance Download PDFInfo
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- CN106011232A CN106011232A CN201610312462.6A CN201610312462A CN106011232A CN 106011232 A CN106011232 A CN 106011232A CN 201610312462 A CN201610312462 A CN 201610312462A CN 106011232 A CN106011232 A CN 106011232A
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- 241000287828 Gallus gallus Species 0.000 title claims abstract description 59
- 238000000034 method Methods 0.000 title claims abstract description 16
- 239000008280 blood Substances 0.000 claims abstract description 6
- 210000004369 blood Anatomy 0.000 claims abstract description 6
- 238000012408 PCR amplification Methods 0.000 claims abstract description 4
- 238000001962 electrophoresis Methods 0.000 claims abstract description 4
- 239000012634 fragment Substances 0.000 claims description 31
- 108010014251 Muramidase Proteins 0.000 claims description 16
- 108010062010 N-Acetylmuramoyl-L-alanine Amidase Proteins 0.000 claims description 16
- 238000003780 insertion Methods 0.000 claims description 13
- 230000037431 insertion Effects 0.000 claims description 13
- 238000009395 breeding Methods 0.000 claims description 8
- 230000001488 breeding effect Effects 0.000 claims description 6
- 238000001514 detection method Methods 0.000 claims description 5
- 230000008030 elimination Effects 0.000 claims description 4
- 238000003379 elimination reaction Methods 0.000 claims description 4
- 230000008034 disappearance Effects 0.000 claims 1
- 238000000605 extraction Methods 0.000 claims 1
- 108090000623 proteins and genes Proteins 0.000 abstract description 17
- 230000004083 survival effect Effects 0.000 abstract description 4
- 230000002350 accommodative effect Effects 0.000 abstract 1
- 238000004458 analytical method Methods 0.000 abstract 1
- 230000012447 hatching Effects 0.000 abstract 1
- 238000009333 weeding Methods 0.000 abstract 1
- 102000016943 Muramidase Human genes 0.000 description 9
- 229960000274 lysozyme Drugs 0.000 description 9
- 235000010335 lysozyme Nutrition 0.000 description 9
- 239000004325 lysozyme Substances 0.000 description 9
- 238000012217 deletion Methods 0.000 description 5
- 230000037430 deletion Effects 0.000 description 5
- 201000010099 disease Diseases 0.000 description 5
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 5
- 101150078994 La gene Proteins 0.000 description 4
- 238000004519 manufacturing process Methods 0.000 description 4
- 235000013601 eggs Nutrition 0.000 description 3
- 238000009394 selective breeding Methods 0.000 description 3
- 241000894006 Bacteria Species 0.000 description 2
- 208000004668 avian leukosis Diseases 0.000 description 2
- 230000033228 biological regulation Effects 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 230000007850 degeneration Effects 0.000 description 2
- 230000036039 immunity Effects 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 230000005570 vertical transmission Effects 0.000 description 2
- QCVGEOXPDFCNHA-UHFFFAOYSA-N 5,5-dimethyl-2,4-dioxo-1,3-oxazolidine-3-carboxamide Chemical compound CC1(C)OC(=O)N(C(N)=O)C1=O QCVGEOXPDFCNHA-UHFFFAOYSA-N 0.000 description 1
- 238000007400 DNA extraction Methods 0.000 description 1
- 102000002322 Egg Proteins Human genes 0.000 description 1
- 108010000912 Egg Proteins Proteins 0.000 description 1
- 108700018351 Major Histocompatibility Complex Proteins 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 241000204031 Mycoplasma Species 0.000 description 1
- 241000202967 Mycoplasma iowae Species 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 238000009360 aquaculture Methods 0.000 description 1
- 244000144974 aquaculture Species 0.000 description 1
- 210000002421 cell wall Anatomy 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000006073 displacement reaction Methods 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 235000014103 egg white Nutrition 0.000 description 1
- 210000000969 egg white Anatomy 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 102000054766 genetic haplotypes Human genes 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 238000002649 immunization Methods 0.000 description 1
- 230000003053 immunization Effects 0.000 description 1
- 230000009545 invasion Effects 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 238000013507 mapping Methods 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 230000013011 mating Effects 0.000 description 1
- 108020004999 messenger RNA Proteins 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 230000001228 trophic effect Effects 0.000 description 1
- 229960005486 vaccine Drugs 0.000 description 1
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- 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
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K67/00—Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals
- A01K67/027—New or modified breeds of vertebrates
- A01K67/0275—Genetically modified vertebrates, e.g. transgenic
- A01K67/0276—Knock-out vertebrates
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/156—Polymorphic or mutational markers
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- 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
- C12Q2600/158—Expression markers
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Abstract
The invention discloses a method for selecting chicken resisting performance. The method includes the steps of extracting chicken blood DNA; conducting PCR amplification by means of two primer pairs, wherein the first primer pair is shown in SEQ ID NO:1 and SEQ ID NO:2, and the second primer pair is shown in SEQ IN NO:3 and SEQ IN NO:4; conducting electrophoresis analysis on a PCR amplification segment to determine the specific gene type; selectively weeding out the LALB gene type and keeping the LALA gene type. In the method, chicken of the LALA gene type are high in hatching rate, survival rate and environment accommodative ability.
Description
Technical field
The invention belongs to aquaculture technical field, relate to chicken breeding technology, be specifically related to a kind of chicken anti-adversity that selects
Method.
Background technology
Chicken suffer climate change, trophic factors change and infect source of disease invasion and attack time, the health level of chicken and the level of production
Can decline, disease can occur time serious, even dead.If chicken group rings border can not be adjusted in time, or takes in time
The measures such as immunity and treatment, the mortality rate of chicken group can be further up, and laying rate declines further.The anti-adversity of chicken is by chicken
Growth level and genetic control.In recent years, the survival level of chicken improves constantly, and the chicken group 72 of major disease does not occurs
Week old survival rate brings up to present 90-95% from 10 years front 85-90%.The raising of scale chicken house adult livability mainly has
The contribution of 3 aspects.One side is that the knowhow producing practitioner of scale chicken house improves, second aspect, socialization
The contribution of the division of labor.This includes many aspects, as strengthened recuperating groupuscule the free compulsory immunization at family in the Yi Kong center of Government-Leading,
Vaccine competition brings better quality, and feedstuff competition brings preferably service, and the fluctuating that the market price is big has eliminated environment relatively
The foster family of difference.The third aspect, is also topmost aspect, is owing to breeding companies has done substantial amounts of in terms of chicken mating system selective breeding
Work.The technology that chicken resistance improves in current breeding companies includes: chicken vertical transmission disease purifies, family selective breeding and labelling auxiliary
Selection-breeding.Chicken vertical transmission disease purifies and mainly purifies S. pullonum and avian leukosis, immunity chicken virus mycoplasma and chicken
M.iowae, by persevering effort in more than 10 years, improves the immune protection of chicken.Family selective breeding is the most logical
Cross the family information utilizing chicken, analyze the death of chicken and eliminate, estimating chicken individuals and the anti-adversity of family, selecting anti-adversity
Good family.Marker assisted selection mainly selects the main histocompatibility complex gene of chicken, as by superseded ev21 haplotype,
Reduce the susceptibility of avian leukosis, thus improve the resistance of chicken.
Lysozyme is the protein that animals and plants is own, the cell wall of this protein energy dissolution of bacteria, thus suppresses antibacterial
Breeding, even eliminating bacteria.Lysozyme is widely used in medical treatment and field of food.But it is degeneration-resistant to utilize lysozyme gene to carry out chicken
The selection-breeding of property it is not yet reported that.
Summary of the invention
It is an object of the invention to provide a kind of method selecting chicken anti-adversity, improve the degeneration-resistant of chicken by gene Selection
Performance.
The present inventor, when studying C type lysozyme, is found that in high-yield egg chicken strain 40 mononucleotides are dashed forward
Displacement point (SNP) and 5 lacking insertions (indel), constitute lysozyme of chicken gene polynorphisms.High-yield egg chicken strain warp
After crossing the selection of more than 10 year, define 2 kinds of genotype, the gene order of these 2 genotype and the chicken base with reference to gene mapping
Because sequence has the difference in up to more than 100 site.In chicken group's genotype after selection, 1 mrna length is than another gene length
4bp, the long named LA of genotype are short for LB.Inventor study find the incubation rate of LA and survival rate higher than LB, and LA
Dominant to LB.Detect the second, the 4th and the 5th insertion and deletion, detect lysozyme gene type.Second insertion and deletion fragment is
30bp ,-4046_-4015delGACAAGTTTATGCATTTTATTACTTCTATT (SEQ ID NO:5), the 4th insertion and deletion
Fragment is 2345_2365delATAGCACAGGGCTTATGCTGC (SEQ ID NO:6), and the 5th insertion and deletion fragment is
2378_2382delTGGA。
It is an object of the invention to be achieved through the following technical solutions:
First aspect, the present invention relates to a kind of method selecting chicken anti-adversity, and described method comprises the steps:
S1, general technical step is used to extract chicken blood DNA；
S2, with described chicken blood DNA as template, use two primers to carrying out PCR amplification；；First primer is to for SEQ
ID NO:1, SEQ ID NO:2；Second primer is to for SEQ ID NO:3, SEQ ID NO:4；
S3, electrophoresis detection pcr amplified fragment: if the first primer is 153bp to pcr amplified fragment, PCR is expanded by the second primer
Increasing fragment is 153bp, then be LALA genotype；If the such as first primer is 183bp to pcr amplified fragment, PCR is expanded by the second primer
Increasing fragment is 128bp, then be LBLB genotype；If the first primer is 153bp to pcr amplified fragment, PCR is expanded by the second primer
Fragment is 128bp, then be LALB genotype；If the such as first primer is 183bp to pcr amplified fragment, PCR is expanded by the second primer
Fragment is 153bp, then be LALB genotype；
S4, superseded LBLB genotype, selective elimination LALB genotype, retain LALA genotype.
Preferably, the first primer as shown in SEQ ID NO:1, SEQ ID NO:2 is to being used for detecting C type lysozyme gene
A lacking insertions, the sequence of this lacking insertions such as SEQ ID NO:5.
Preferably, the second primer as shown in SEQ ID NO:3, SEQ ID NO:4 is to being used for detecting C type lysozyme gene
Two lacking insertions, the sequence of said two lacking insertions such as SEQ ID NO:6 and " TGGA ".
Preferably, the sequence of LA gene is as shown in SEQ ID NO:7.
Preferably, the sequence of pcr amplified fragment fragment after gene sequencing is as shown in SEQ ID NO:8.
Preferably, in addition to above-mentioned Insert Fragment, the regulation and control region of LA gene also has the spy as shown in SEQ ID NO:9
Levy sequence.
Preferably, in step S4, selective elimination LALB genotype is to eliminate depending on group size, particularly as follows: group
In body, when LALA genotype hen group is more than 300, eliminate LALB genotype hen；When LALA genotype cock is more than 80,
Eliminate LALB genotype cock.
Second aspect, the invention still further relates to a kind of aforesaid method purposes in chicken breeding selecting chicken anti-adversity.
Preferably, when pure lines are reserved seed for planting, select LALA genotype cock to breed with LALA genotype hen, make progeny population complete
Entirely become LALA genotype.
Compared with prior art, there is advantages that
1, the present invention is by detecting the genotype of lysozyme of chicken gene, selects the gene of the strong stress resistance of lysozyme of chicken gene
Type, thus improve the anti-adversity of chicken；
2, the present invention screens the chicken with LALA genotype of acquisition, and incubation rate is high；
3, the present invention screens the adult livability height with LALA genotype of acquisition；
4, the present invention selects to obtain pure LALA genotype chicken colony is suitable for environment than LBLB genotypic population；
5, the present invention selects the pure LALA genotype hen obtained to contain more than in the egg of LBLB genotype hen
Lysozyme, i.e. egg white lysozyme content are high；
6, the production performance of the chicken with LALA genotype that the present invention screens acquisition is consistent with LBLB.
Detailed description of the invention
Below in conjunction with embodiment, the present invention is described in detail.Following example will assist in those skilled in the art
It is further appreciated by the present invention, but limits the present invention the most in any form.It should be pointed out that, to those of ordinary skill in the art
For, without departing from the inventive concept of the premise, it is also possible to make certain adjustments and improvements.These broadly fall into the guarantor of the present invention
Protect scope.
Embodiment
1, design 2 is to primer, as shown in table 1 below:
Table 1
2, use general DNA extraction technology and PCR detection technique, extract the blood DNA of chicken, use the DNA designed
Primer sequence, uses the chicken DNA of known LALA and LBLB to do reference, detection sequence 1 and fragment (the i.e. C type lysozyme base of sequence 2
3 insertion and deletion fragments in Yin).
3, the genotype of the C type lysozyme gene of individuality is determined.Particularly as follows: electrophoresis detection pcr amplified fragment, 2 genes
Fragment such as table 2 below, according to the size of fragment, determine and belong to LA or LB genotype；
Table 2
| Gene | LA | LB |
| First pair of primer PCR product | 153bp | 183bp |
| Second pair of PCR primer | 153bp | 128bp |
In the present invention, the sequence of LA gene is as shown in SEQ ID NO:7；Above-mentioned pcr amplified fragment is through base
Because the sequence of the fragment after order-checking is as shown in SEQ ID NO:8.In addition to above-mentioned Insert Fragment, LA gene
Regulation and control region also has the characteristic sequence as shown in SEQ ID NO:9.
The production performance such as table 3 below of LALA, LALB and LBLB genotype chicken obtained:
The production performance of 33 kinds of genotype of table
4, as found that there is LBLB genotype in colony, this genotype is eliminated；As colony exists LALA genotype, retain
LALA genotype.For LALB genotype, eliminate depending on group size, during particularly as follows: LALA group is more than 300 hens, wash in a pan
Eliminate LALB hen；When LALA cock is more than 80, LALB cock can be eliminated.
5, when pure lines are reserved seed for planting, select LALA cock to breed with LALA hen, make progeny population go completely into LALA genotype.
Claims (6)
1. the method selecting chicken anti-adversity, it is characterised in that described method comprises the steps:
S1, extraction chicken blood DNA；
S2, with described chicken blood DNA as template, use two to primer to carrying out PCR amplification；First primer is to such as SEQ ID NO:
Shown in 1, SEQ ID NO:2；Second primer is to as shown in SEQ ID NO:3, SEQ ID NO:4；
S3, electrophoresis detection pcr amplified fragment: if the first primer is 153bp to pcr amplified fragment, the second primer expands sheet to PCR
Section is 153bp, then be LALA genotype；If the such as first primer is 183bp to pcr amplified fragment, the second primer expands sheet to PCR
Section is 128bp, then be LBLB genotype；If the first primer is 153bp to pcr amplified fragment, the second primer is to pcr amplified fragment
For 128bp, then it it is LALB genotype；If the such as first primer is 183bp to pcr amplified fragment, the second primer is to pcr amplified fragment
For 153bp, then it it is LALB genotype；
S4, superseded LBLB genotype, selective elimination LALB genotype, retain LALA genotype.
The method selecting chicken anti-adversity the most according to claim 1, it is characterised in that such as SEQ ID NO:1, SEQ ID
The first primer shown in NO:2 to for detecting a lacking insertions of C type lysozyme gene, this lacking insertions
Sequence such as SEQ ID NO:5.
The method selecting chicken anti-adversity the most according to claim 1, it is characterised in that such as SEQ ID NO:3, SEQ ID
Two lacking insertions for detecting C type lysozyme gene, said two disappearance are inserted by the second primer shown in NO:4
The sequence of fragment such as SEQ ID NO:6 and " TGGA ".
The method selecting chicken anti-adversity the most according to claim 1, it is characterised in that in step S4, selective elimination
LALB genotype is to eliminate depending on group size, particularly as follows: in colony, when LALA genotype hen group is more than 300, wash in a pan
Eliminate LALB genotype hen；When LALA genotype cock is more than 80, eliminate LALB genotype cock.
5. the method for a selection chicken anti-adversity as claimed in claim 1 purposes in chicken breeding.
Purposes the most according to claim 5, it is characterised in that when pure lines are reserved seed for planting, selects LALA genotype cock and LALA
Genotype hen breeds, and makes progeny population go completely into LALA genotype.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610312462.6A CN106011232B (en) | 2016-05-12 | 2016-05-12 | A method of selection chicken anti-adversity |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610312462.6A CN106011232B (en) | 2016-05-12 | 2016-05-12 | A method of selection chicken anti-adversity |
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| Publication Number | Publication Date |
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| CN106011232A true CN106011232A (en) | 2016-10-12 |
| CN106011232B CN106011232B (en) | 2019-10-11 |
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103602745A (en) * | 2013-11-20 | 2014-02-26 | 河南农业大学 | Primers, kit and detection method for detecting gene type of dominant white feather site of chicken PMEL17 gene |
| CN104293905A (en) * | 2014-04-11 | 2015-01-21 | 河南农业大学 | Primer, kit and detection method for detecting chicken green shin character linkage SNP locus genotype |
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- 2016-05-12 CN CN201610312462.6A patent/CN106011232B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103602745A (en) * | 2013-11-20 | 2014-02-26 | 河南农业大学 | Primers, kit and detection method for detecting gene type of dominant white feather site of chicken PMEL17 gene |
| CN104293905A (en) * | 2014-04-11 | 2015-01-21 | 河南农业大学 | Primer, kit and detection method for detecting chicken green shin character linkage SNP locus genotype |
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| Title |
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| 佚名: "登录号：XR_001465456", 《GENBANK》 * |
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Address after: 201403 No. 2901 Zhai Road, Minhang District, Shanghai Patentee after: Shanghai Academy of Agricultural Sciences Address before: 201403, No. 1000, Qi Lu, Shanghai, Fengxian District Patentee before: Shanghai Academy of Agricultural Sciences |