Chicken growth and development related InDel polymorphism marker, primer, kit and application

Abstract

The present invention belongs to the field of biomolecular markers, and specifically relates to an InDel polymorphism marker, primer, kit and application related to chicken growth and development. The InDel polymorphism marker is a 27bp insertion or deletion fragment in the sixth intron region of the chicken YBX3 gene, and the nucleotide sequence of the insertion or deletion fragment is shown in SEQ ID NO.6. The present invention first detected the distribution of the 27bp InDel polymorphism marker in the sixth intron region of the YBX3 gene in commercial broilers, Chinese local chickens and high-yielding laying hens, and analyzed its genotype distribution and allele frequency in different breeds of chickens. Subsequently, the mutation polymorphism was associated with the economic traits of the Gushi chicken × Anka chicken F2 resource population, and it was found that the polymorphism marker was closely related to the chicken growth traits and 84-day slaughter traits, which laid the foundation for screening out homozygous high-quality local chicken populations.

Description

Technical Field

[0001] The invention belongs to the field of biomolecular markers, and specifically relates to a chicken growth and development-related InDel polymorphism marker, a primer, a kit and an application. Background Art

[0002] The slow growth rate of local chicken breeds in China has always been a problem faced in the breeding process. The selection and breeding of commercial broilers and laying hens abroad has a long history. Whether they have differences in the genes related to growth control from local chicken breeds in China is still poorly understood. Therefore, it is very important and necessary to use molecular biological methods to diagnose the genotype of a gene related to body weight in an individual for breeding. Molecular markers are a type of genetic marker that can reflect individual specificity by directly performing genetic analysis at the nucleic acid level. The use of molecular marker technology helps to reveal genetic variation and its variation patterns in essence. Molecular markers have been widely studied and applied in chicken genetic analysis, population genetics research, gene positioning and breeding. In order to accelerate the genetic improvement of broiler and local chicken breeds, combining advanced molecular marker technology with traditional breeding is an important means of poultry breeding development.

[0003] Insertion / Deletion (InDel) markers developed based on whole genome resequencing refer to the differences between two parents in the whole genome sequence. InDel markers have attracted more and more attention due to their advantages such as wide distribution in the genome, high density, stable variation, strong polymorphism, and easy detection. They have important application value in biological genetic diversity, variety purity identification, kinship identification, and molecular breeding.

[0004] The detection of human markers has become an important means of genetic diagnosis. With the development of InDel markers located on functional genes, combined with chromosome walking and gene fine positioning, these markers can be used to screen functional genes for economic traits of agricultural animals, which is conducive to the further development and utilization of excellent genes. Summary of the invention

[0005] The purpose of the present invention is to provide an InDel polymorphic marker related to chicken growth and development, which has a high correlation with the growth performance and carcass traits of chickens.

[0006] The second object of the present invention is to provide diagnostic primers and kits for the above-mentioned chicken growth and development-related InDel polymorphism markers.

[0007] The third object of the present invention is to provide the application of the above-mentioned markers, diagnostic primers and kits.

[0008] In order to achieve the above purpose, the technical solution adopted by the present invention is:

[0009] A chicken growth and development related InDel polymorphism marker, wherein the InDel polymorphism marker is a 27 bp insertion or deletion fragment in the sixth intron region of the chicken YBX3 gene, and the nucleotide sequence of the insertion or deletion fragment is shown in SEQ ID NO.6.

[0010] The chicken growth and development related InDel polymorphic marker of the present invention is located in the sixth intron of the YBX3 gene, and the 27bp insertion occurs between 17898-17899 of the YBX3 gene. The NCBI GenBank accession number of the chicken gene sequence is NC_006088.5. The present invention first detected the distribution of the 27bp InDel polymorphic marker in the sixth intron of the YBX3 gene in commercial broilers, Chinese local chickens and high-yielding laying hens, and analyzed its genotype distribution and allele frequency in different breeds of chickens. Subsequently, the mutation polymorphism was associated with the economic traits of the Gushi chicken × Anka chicken F2 resource population, and it was found that the polymorphic marker was closely related to the chicken growth traits and 84-day slaughter traits, which laid the foundation for screening out homozygous high-quality local chicken populations.

[0011] Preferably, the sequence of the sixth intron region of the chicken YBX3 gene containing the 27 bp insertion is as shown in SEQ ID NO. 1, and the insertion site is located at position 934-960 from the 5′ end.

[0012] A primer for detecting the above-mentioned InDel polymorphic marker.

[0013] Corresponding primers are designed based on the InDel polymorphic marker, and chicken individuals with homozygous insertion of 27 bp can be detected and screened. Therefore, the primers can be used for auxiliary selection and molecular breeding of chickens.

[0014] Preferably, the amplified fragment of the primer covers the insertion or deletion fragment region.

[0015] Further preferably, the primers include an upstream primer having a nucleotide sequence as shown in SEQ ID NO.2, and a downstream primer having a nucleotide sequence as shown in SEQ ID NO.3.

[0016] A kit containing the above primers.

[0017] Based on the above primers, other reagents known in the art can be used to construct a kit, so as to conveniently and quickly achieve genotype determination, and can be further used for assisted selection and molecular breeding of chickens. The kit may include one or more of dNTPs, PCR reaction buffer, DNA polymerase and DNA Marker. These reagents in the kit are commonly used reagents in the PCR amplification process.

[0018] By designing corresponding primers and kits based on the InDel polymorphism marker, chicken individuals with homozygous insertion of 27bp can be detected, and then individuals with fast growth rate can be quickly screened for molecular breeding, thereby shortening the breeding time and accelerating the breeding process.

[0019] The application of the above-mentioned InDel polymorphism marker, primer, and kit in predicting the growth status of a chicken flock comprises the following steps: designing primers based on the InDel polymorphism marker and performing PCR amplification, judging the genotype of the chicken to be tested as homozygous insertion type II, homozygous deletion type DD, or heterozygous type ID according to the PCR amplification result, and predicting the growth status of the chicken flock according to the genotype determination result.

[0020] Application of the above-mentioned InDel polymorphism markers, primers and kits in assisted selection and molecular breeding of chickens.

[0021] Preferably, primers are designed based on the InDel polymorphic marker and PCR amplification is performed. The genotype of the chicken to be tested is judged to be homozygous insertion type II, homozygous deletion type DD or heterozygous type ID based on the PCR amplification results. The growth traits and / or slaughter traits of individuals with genotype II are the best, and homozygous insertion type II individuals are selected for molecular breeding.

[0022] Further preferably, the growth traits are chicken weight, shank circumference, chest width, sternum length, pelvic width, and body oblique length; and the slaughter traits are half-eviscerated weight, full-eviscerated weight, half-eviscerated rate, full-eviscerated rate, head weight, claw weight, wing weight, gizzard weight, pancreas weight, breast muscle weight, leg weight, leg muscle weight, carcass weight, and chest circumference. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 This is a schematic diagram of the technical process of breeding using InDel polymorphic markers in the present invention;

[0024] Figure 2 The figure is an agarose gel electrophoresis diagram of the chicken YBX3 gene amplification product in the embodiment of the present invention. DETAILED DESCRIPTION

[0025] YBX3 is a DNA / RNA binding protein associated with gene transcription, DNA repair and the progression of various diseases. It has a cold shock domain and is highly conserved from bacteria to humans. Previous studies have shown that the YBX3 gene is ubiquitously expressed in human skeletal muscle, heart and decidual cells, and plays an important role in the transcription and translation of target genes involved in cell survival, proliferation and differentiation. However, the effects of the YBX3 gene and its mutation on poultry growth traits have rarely been reported.

[0026] The present invention provides a theoretical basis for chicken molecular breeding by identifying the sequence variation of the YBX3 gene and exploring whether the YBX3 gene can be used as a molecular marker to select poultry weight indicators or other production traits.

[0027] In the present invention, the study found that the 27bp InDel polymorphic marker in the sixth intron region of the chicken YBX3 gene was significantly correlated with a variety of growth traits, specifically: the mutation of 27bp in the sixth intron region of the YBX3 gene was significantly correlated with the weight of chickens at 0, 2, 4, 6, 10, and 12 weeks, shank circumference at 4 and 8 weeks, chest width at 4 and 12 weeks, sternum length at 4, 8 and 12 weeks, pelvic width at 4 and 8 weeks, and body oblique length at 4 weeks (P<0.05). In addition, the InDel was also significantly correlated with 84-day slaughter traits, specifically: half-eviscerated weight, full-eviscerated weight, half-eviscerated rate, full-eviscerated rate, head weight, claw weight, wing weight, gizzard weight, pancreas weight, breast muscle weight, leg weight, leg muscle weight, carcass weight, and chest circumference (P<0.05).

[0028] Corresponding primers are designed based on the InDel polymorphic marker, and chicken individuals with homozygous insertion of 27 bp can be detected and screened. Therefore, the primers can be used for auxiliary selection and molecular breeding of chickens.

[0029] Specifically, the primer sequence is as follows:

[0030] PF: 5′-GGCTAGACAACCCCTACTCTC-3′;

[0031] PR: 5′-ATGGCCATCAGGTGGTGATT-3′.

[0032] The amplified fragment of the primer covers the 17876-18041th position from the 5' end of the sixth intron region of the chicken YBX3 gene, and the NCBI GenBank accession number of the chicken gene sequence is NC_006088.5. If it is a 27bp insertion genotype, the size of the PCR amplified fragment is 193bp, and if it is a 27bp deletion genotype, the size of the PCR amplified fragment is 166bp.

[0033] The method for detecting the sixth intron genotype of the chicken YBX3 gene using the above primers is as follows:

[0034] 1) Design primers based on the sequences before and after the 27 bp insertion / deletion site in the sixth intron of the chicken YBX3 gene, and the amplified fragments of the primers cover the 27 bp insertion / deletion fragment region in the sixth intron of the chicken YBX3 gene. Specifically, the amplified fragments of the primers cover the 17876-18041 positions from the 5′ end of the sixth intron region of the chicken YBX3 gene, and the NCBI GenBank accession number of the chicken gene sequence is NC_006088.5. The nucleotide sequence of the 27 bp insertion or deletion fragment in the sixth intron of the chicken YBX3 gene is shown in SEQ ID NO.6;

[0035] Extract the genomic DNA of the chicken to be tested, and perform PCR amplification using the designed primers;

[0036] 2) Determine the genotype of the chicken to be tested based on the amplification results.

[0037] The above method greatly improves the efficiency and accuracy of genotype determination, and the method is simple, the typing time is short, sequencing and restriction endonucleases are not required, special instruments are not required, the cost is low, and it is easy to promote and popularize. The method can quickly determine the genotypes of different weight traits, thereby shortening the breeding time and accelerating the breeding process. The method of the present invention detects the distribution of InDel polymorphic markers in the sixth intron region of the chicken YBX3 gene in local chickens, high-yield laying hens and commercial broilers, and analyzes the distribution of genotypes and allele frequencies of InDel polymorphic markers in different breeds of chickens.

[0038] The genotype is homozygous insertion type II, homozygous deletion type DD or heterozygous type ID, wherein the I allele is expressed as a 27bp insertion and the D allele is expressed as a 27bp deletion mutation. If the PCR amplified fragments all contain this 27bp fragment, the chicken to be tested is of II genotype and the individual is a homozygous body; if the amplified fragments do not contain this 27bp fragment, the chicken to be tested is of DD genotype and the individual is a homozygous body; if some of the amplified fragments contain this 27bp fragment and some do not, the chicken to be tested is of ID genotype and the individual is a heterozygous body.

[0039] The above genotype determination results can be used to predict the growth status of the chicken flock.

[0040] The present invention has found through research that the growth trait association analysis value of II genotype individuals is the largest, the association analysis value of ID genotype individuals is the smallest, and the association analysis value of DD genotype individuals is in the middle; II genotype individuals have the best growth condition, DD genotype individuals are the second, and ID genotype individuals are the worst. According to the number of II, ID, and DD genotype individuals in the flock, the growth condition of the flock is predicted.

[0041] The above genotype determination results can be used to screen out homozygous insertion type II individuals for breeding.

[0042] The present invention found through research that the growth trait association analysis value of II genotype individuals is the largest, the association analysis value of ID genotype individuals is the smallest, and the association analysis value of DD genotype individuals is in the middle. Therefore, the economic performance of chickens can be improved by strengthening the selection and breeding of II genotype individuals.

[0043] The present invention is further described in detail below in conjunction with specific examples. Unless otherwise specified, the equipment and reagents used in each example are conventionally commercially available.

[0044] Example 1 InDel polymorphic markers related to chicken growth and development

[0045] The chicken growth and development related InDel polymorphic marker of this embodiment is a 27 bp insertion or deletion fragment in the sixth intron region of the chicken YBX3 gene, and the nucleotide sequence of the insertion or deletion fragment is shown in SEQ ID NO. 6. The sequence of the sixth intron region of the chicken YBX3 gene containing the 27 bp insertion is shown in SEQ ID NO. 1, and the insertion site is located at positions 934-960 from the 5′ end.

[0046] Example 2 Primers, kit

[0047] The primers of this example are designed for the InDel polymorphism marker of Example 1, and the amplified fragment of the primer covers the 27 bp insertion or deletion fragment region in the sixth intron of the chicken YBX3 gene; specifically, the primers are designed according to the YBX3 gene sequence (GenBank Accession NC_006088.5) published by NCBI, the sequence of the sixth intron region of the YBX3 gene is shown in SEQ ID NO.1, and the primer sequence is shown as follows:

[0048] PF: 5′-GGCTAGACAACCCCTACTCTC-3′ (as shown in SEQ ID NO. 2);

[0049] PR: 5′-ATGGCCATCAGGTGGTGATT-3′ (as shown in SEQ ID NO. 3).

[0050] The size of the fragment amplified by the above primers is 193 bp (as shown in SEQ ID NO.4; corresponding to the 911-1103th position of the sixth intron region sequence of the YBX3 gene as shown in SEQ ID NO.1) or 166 bp (as shown in SEQ ID NO.5), 193 bp is the I allele, and 166 bp is the D allele. There is a 27 bp insertion in the I allele, and the nucleotide sequence of the inserted fragment is shown in SEQ ID NO.6.

[0051] The kit of this embodiment contains the above-mentioned primers of this embodiment, and other conventional components, such as dNTPs, sterile ultrapure water, PCR reaction buffer, DNA polymerase and DNA Marker.

[0052] The method for detecting the genotype of the sixth intron region of the chicken YBX3 gene using the above primers or kit is as follows:

[0053] (1) Extracting genomic DNA from the chicken to be tested and performing PCR amplification using the designed primers;

[0054] The genomic DNA of the tested chickens was extracted using the phenol-chloroform crude extraction method.

[0055] The reaction procedure of PCR amplification was as follows: pre-denaturation at 95°C for 5 min; denaturation at 95°C for 30 s, annealing at 60°C for 30 s, extension at 72°C for 30 s, 30 cycles; extension at 72°C for 10 min; and storage at 4°C.

[0056] The reaction system for PCR amplification was 3.0 μL of 2×Taq PCR MasterMix, 0.5 μL of primer PF (10 pmol / L), 0.5 μL of primer PR (10 pmol / L), 1.0 μL of genomic DNA of the chicken to be tested (50 ng / μL), and 5.0 μL of sterile ultrapure water.

[0057] (2) Determine the genotype of the chicken to be tested based on the amplification results. The genotype is divided into homozygous insertion type II, homozygous deletion type DD or heterozygous type ID, where the I allele shows a 27 bp insertion mutation and the D allele shows a 27 bp deletion mutation.

[0058] Example 3 Application in predicting chicken flock growth status

[0059] In this embodiment, based on the embodiment 2, the growth status of the chickens is further predicted according to the genotype determination results.

[0060] The association analysis value of growth traits and slaughter traits of individuals with II genotype is the largest, the association analysis value of individuals with ID genotype is the smallest, and the association analysis value of individuals with DD genotype is in the middle; individuals with II genotype have the best growth condition, followed by individuals with DD genotype, and individuals with ID genotype are the worst. The growth condition of the flock can be predicted based on the number of individuals with II, ID, and DD genotypes in the flock.

[0061] Growth conditions include body weight at 0, 2, 4, 6, 10, and 12 weeks, shank circumference at 4 and 8 weeks, chest width at 4 and 12 weeks, sternum length at 4, 8, and 12 weeks, pelvic width at 4 and 8 weeks, body oblique length at 4 weeks, half-eviscerated weight, full-eviscerated weight, half-eviscerated rate, full-eviscerated rate, head weight, claw weight, wing weight, gizzard weight, pancreas weight, breast muscle weight, leg weight, leg muscle weight, carcass weight, chest circumference, etc.

[0062] Example 4 Application in assisted selection and molecular breeding of chickens

[0063] In this example, based on Example 2, homozygous insertion type II individuals were further screened out according to the genotype determination results for breeding.

[0064] Experimental Example 1

[0065] In this experimental example, the correlation between the insertion or deletion of 27bp in the sixth intron of the chicken YBX3 gene and the growth status of chickens was studied. The dominant genotype was identified and individuals with these dominant genotypes were selected for breeding. The overall technical flow chart is shown below: Figure 1 As shown, the primers used in the PCR reaction in the study are the same as the primers in Example 2, and the research process includes the following steps:

[0066] (1) Sample collection

[0067] 684 F2 generation resource groups (Gushi chicken × Anka) of Henan Agricultural University, 521 AA broilers, 279 Hubbard broilers, 120 Cobb broilers, 155 Ross 308 broilers, 64 817 broilers, 260 Gushi chickens, 109 Xichuan black-bone chickens, 184 Lushi chickens, 230 Changshun green-shell laying hens, 308 Dongxiang green-shell laying hens, 339 Wuhei chickens, 314 Hailan brown-shell laying hens, and 44 Roman laying hens were collected. Blood was collected from the wing vein, 1 / 3 anticoagulant was added, and genomic DNA was extracted by phenol-chloroform crude extraction method and stored in a 4°C refrigerator for later use.

[0068] (2) PCR amplification

[0069] The PCR reaction system adopted the mixed addition method, that is, according to the quantity of various components required for each reaction system and the number of PCR reactions required for one reaction, the total amount of various reaction components was calculated, added to a 1.5 mL centrifuge tube, fully mixed and centrifuged instantly, and then respectively packaged into 0.2 mL Eppendorf PCR tubes, and then template DNA was added respectively, and then PCR amplification was performed after instantaneous centrifugation; the PCR reaction system is shown in Table 1, among which 2×Taq PCR MasterMix was purchased from Nanjing Novozyme Biotechnology Co., Ltd.

[0070] The PCR reaction program was as follows: pre-denaturation at 95°C for 5 min; denaturation at 95°C for 30 s, annealing at 60°C for 30 s, extension at 72°C for 30 s, 30 cycles; extension at 72°C for 10 min; and storage at 4°C.

[0071] Table 1 PCR reaction system

[0072] <![CDATA[Sterilized ultrapure water (H2O)]]> 5.0μL 2×Taq PCR MasterMix 3.0μL Primer PF (10 pmol / L) 0.5μL Primer PR (10pmol / L) 0.5μL DNA of the chicken to be tested (50ng / μL) 1.0μL Total volume 10.0μL

[0073] (3) Detection of PCR amplification products and determination of genotypes

[0074] Take 8 μL of PCR amplification product, spot it on 2% agarose gel electrophoresis (electrophoresis voltage: 120V; electrophoresis time: 30min), and detect it with gel imaging system. The electrophoresis pattern results are as follows: Figure 2 As shown, the lanes from left to right represent the electrophoretic band patterns of ID, DD, II, DD, II, ID genotypes and DNA Marker Ⅰ, respectively.

[0075] The method for determining the genotype is as follows: if the PCR amplification product has only a 193bp fragment, the chicken to be tested is of the II genotype and the individual is homozygous; if the amplified fragment size is only 166bp, the chicken to be tested is of the DD genotype and the individual is homozygous; if the amplified fragment size is 193bp and 166bp, the chicken to be tested is of the ID genotype and the individual is heterozygous.

[0076] The genotyping results of the F2 resource group, AA broiler, Hubbard broiler, Cobb broiler, Ross 308, 817 broiler, Gushi chicken, Xichuan black-bone chicken, Lushi chicken, Changshun green-shell laying hen, Dongxiang green-shell laying hen, Wuhei chicken, Hy-Line brown-shell laying hen and Roman laying hen are shown in Table 2.

[0077] Table 2 Statistics of genotype detection results of different groups

[0078]

[0079] (4) Association analysis between growth traits of F2 resource population and genotype of the sixth intron region of YBX3 gene

[0080] There are 684 individuals in the F2 resource population with complete economic trait records for association analysis of the polymorphic locus. The economic trait determination method is as follows: body weight is measured every 2 weeks, body size is measured every 4 weeks, and the chickens are slaughtered at 12 weeks of age. The feed is stopped for 12 hours before slaughter (without stopping water) and then weighed. A total of 57 growth, slaughter and meat quality indicators were measured. The economic trait determination method reference (Han RL, Li ZJ, Li M J. Novel 9-bp InDel in visfatin gene and its associations with chicken growth. Br Poult Sci, 2011, 52(1): 52-57.).

[0081] SPSS 26.0 software was used to analyze the correlation between gene loci and economic traits. The data of each trait were ensured to be normally distributed, and then the least squares method was used to correct the data. According to the data characteristics, the multivariate linear model was used to analyze the genotype effect and the Benferroni multiple comparison method was used to compare the differences between the genotypes. The results are shown in Tables 3 and 4.

[0082] Table 3 Association analysis between 27bp polymorphism of YBX3 gene and growth traits of chicken in F2 resource population

[0083]

[0084] Note: a,b The same superscripts in the same row indicate no significant difference, while different superscripts indicate significant difference.

[0085] Table 4 Association analysis between 27bp polymorphism of YBX3 gene and slaughter traits of chickens in F2 resource population

[0086]

[0087]

[0088] Note: a,b The same superscripts in the same row indicate no significant difference, while different superscripts indicate significant difference.

[0089] The results of association analysis in Tables 3 and 4 show that a 27 bp sequence inserted in the sixth intron of the YBX3 gene, i.e., the InDel mutation polymorphism in the sixth intron of the YBX3 gene, is significantly associated with body weight at 0, 2, 4, 6, 10, and 12 weeks, shank circumference at 4 and 8 weeks, chest width at 4 and 12 weeks, sternum length at 4, 8, and 12 weeks, pelvic width at 4 and 8 weeks, and body oblique length at 4 weeks (P<0.05). In addition, the InDel is also significantly associated with 84-day slaughter traits, specifically: half-eviscerated weight, full-eviscerated weight, half-eviscerated rate, full-eviscerated rate, head weight, claw weight, wing weight, gizzard weight, pancreas weight, breast muscle weight, leg weight, leg muscle weight, carcass weight, and breast circumference (P<0.05).

[0090] And overall, the growth trait association analysis value of II genotype individuals was the largest, while that of ID genotype individuals was the smallest, indicating that the 27bp insertion in the sixth intron region of the YBX3 gene was beneficial to the growth and development of chickens.

[0091] On this basis, the tested individuals are bred to select excellent varieties. Specifically, the selection and breeding of genotype II individuals is strengthened to improve the economic performance of chickens; and the ID and DD genotype individuals are eliminated. <110> Henan Agricultural University <120> Chicken growth and development related InDel polymorphism marker, primer, kit and application <160> 6 <170> PatentIn version 3.5 <210> 1 <211> 5566 <212> DNA <213> Chicken <221> Sequence of the sixth intron region of the YBX3 gene <400> 1 gtttgtgctg tttctttgct catcattgca aatgagtata gtctgtaaaa gcttgaggag 60 tataaagctg aattcccaga cattcaagta catgcaagtg gtggagagaa gtgtttaaat 120 tacctggagg aaggtgtaac ttctaggagt aaggaaatag ttgtagaagt tgcttgatct 180 aattgtacgt tactcagatc ctacttgaca tggggcaggt tggtccagcg tctgcagggc 240 caggatgtgc tttgaaaagc ttcctgtgga ctgtggccaa tagtctagca ctgtgttgca 300 aactagtggc ttgttcccag gaccaagatg gatctatcac tcaagcatag gtctaagcac 360 tgatgctgct tgtatgtaaa aaagagcctg tttttcagaa ataacataaa gtcatgttct 420 gtaggacaga tcatcagata catggcagga tttcagttac cttgtagtat gctgtgcttg 480 gaagtattgc aattgtgcgt gaccaagggc tatgaggctt ggggaggtgg gagttagggg 540 agggggaggg agggaatatc ttaaaatcat aaagtagttt gggttggaag gcacctttta 600 gaggtcatct agttctaaca accctgacag gagcacttcc tactcagtca ggttgctcaa 660 agccccatct gggcatttgt gaacattttc agggatgggg cttccacaac ctctctaggc 720 aacctgttcc agtgcctcac tgcccacata gtaaaaaaaa aaattttact taaatctact 780 ctttgattta aagccattat tctttgtctt atcactgcat gccctagtaa aaagtctgtc 840 ttcagctttc ctggaagcta ccctttaggt actggaagtt tagtatcagg tctccctgga 900 gtcttctcca ggctagacaa cccctactct ctcccttctt tacaggaaag gtgctccagt 960 agctccagcc ctctgataac attcatggcc cacatctgga cttgctccaa aacatccatg 1020 gtgttatgct caggtcccag ggagcagcac aatactctag gtggggtctg agcagagcag 1080 ggaaatcacc acctgatggc catgcttctt ttgatttagc ccaggatcca gttagctttc 1140 taagctgcaa ctatactgtc agcttatatt catctcttca tccactaaca ccaccaagtc 1200 cttggtgggt ctgctgtcag tccacttact gtctagttgg tattcatgtt tgggattgcc 1260 ccaacccatg tgcaagactt tgtacatggc cttgttgaac ttcatgaggt tctcacctct 1320 caaatctgtt agggttcctc tggaaggcat ttcttccctc cagtgtcagc tacaccactt 1380 accttggtgt catctgcgaa cttactgagg atgtactcaa tgctttgtct gtgccactga 1440 caaagataaa tagtgccagt cccagtatca gtcactgagg aatgcctcgt cactgatctc 1500 taccttgaca tcaagttatt gacatcaagt tactgactgc aactgtttga gcagcaccat 1560 gcaatttctt atccactgag tgacttttta cattggtaga tagtgatagg acgaggagga 1620 atggttttaa actaaaggag gggagattta gataagatgt ccaggggagt ctttcataga 1680 gagggtggtg aggtacttga acaagctgtt gagagaggtt gtggatgttc catccctgga 1740 ggtattcaag gccaggttgg ttggaaaccc tgcttatggc agagggattg aaactagatg 1800 atctttggct tgggttggaa gggacttcat tctgtgaaat acaatggaac tgtttcttag 1860 taacttcatc tgctagagtt tccttgggac ccttaggtgc atcttatcag gtcttgtggg 1920 cttgtgcaac ttcaggttcc ttaggtggtg tctcaaactt gatctcctcc tacagtgggt 1980 ggttcttaat tcttccagtc cctgttttct gtgagttggg cagtatggct atagcacttt 2040 ccaatgaaga ttaaggagga aaaaaaatta tggaatacct cagccttctc catattctgg 2100 ataacagggt ctcctatttt cttacaaaga gggaccacat ttttcctagt ttccttctat 2160 cactgacata cttaaaagcc tttcccattg ctcttgatgt ccctgatcag attcagttat 2220 tctggggctt ttgcttttct aatctgattc ctgactgtgc agacgatgac tgtgttcctt 2280 ccaggttggt tccatccttg tttctggtct ccatatgctt cctttttgtg tttgggtttg 2340 tctcggagct tctcgtattt ttgcctgact tctttttagt tgccttgctc ttgagcttat 2400 aggtgattat tgaatattaa ctgacttttc tgggcctctt ttctttccag ggctctatgc 2460 cctagttctc cacaaggcag gttcctgaga tctgttctcc agaatggtag tgagtttgct 2520 gtgtgccctc cttgttgtca taatcatctc aaactctacc attttgttgt cactacagtc 2580 agtcatagaa tcatcaaggt tggaaaagac ttctcaggat catccagtcc aactgtccac 2640 ctatcaccag tatttccttg tccctcagta caacttctag acatttcttg aacacctcca 2700 aggatggtga ctcccacctc tgggcagcct gtcccagcac ctgactactt tcttgaagta 2760 tttccttgcg tccaacttga atcttcctct ggcacaactt gaggctgttc cctcttgtct tactgcagtg ctgtccttga gcttcgtgtt tctcaccagg ccttccttgt tggtgagaac 2880 aggatccagc gtagcacctg tcctccttgg ctcctctgta tctttaaagg aagctatgat 2940 cagcgcattt ctgagcctcc tggattatgc cttgtgttgt ctgtccaaca gatatcagga 3060. cccccacgaa gatcagggct tgctgaacat gagggtgctc ctttctgtct acagagggcc taatctactc agtcttcgta gttgggtgcc tttagtagtc ccactgtaat 3180. gtgagctgtt gttgtcctcc cttaaatcct gatccagaag ctcttggtca gctccttatc catccccagg tgtagctctg tgtattccag ctggaggttg acttggaggg aatgcttcct 3240 ccttgtctgc tgtttgtttt tcctgaagag cccatatcct tccatttcag ggccataata 3300 atcttatgtg cacaactaac tcctcttgtt tattctccat gcttcatctt tgtatagaga catttaactt gggcccttaa caaacggact tactgcctgg tatggctgaa actcacttat gctgctcatc aagtgttctc cctttgacct gtggtcctgt tttctcccag gttttcagaa 3480 tcatgaatgg catgagaaga atagatgaag tttcattatt tcatttctat tttgttctga 3540 tgtaggaaca ggctatgagg agacatcaaa tgaaggaagt cagacgtgtt ggggggaaaa 3600 aagttatgct tgaagcatat agttagaact ttttgctgca ggatgttggg gagatgcaag 3660 cttgtgtgag cttgagtgag atgggtcaca tagccagaag agaacacttg tatattatt 3720 aaatatgcaa actgtatcta gctcgggaaa tctccagagt ggaaaacaac tggacattag 3780 gagactacct aggggaaata ggcttcttta tccttacgct tccctaagga tccatggaac 3840 tgggttaaag gagtccttga acacccagca ggacttctta agaaaaattt aactaaaaat 3900 tatttggctt ttacatgaac acaaatactt tccaatagga ttagctgtgt aatttgctga 3960 gtcgcctatg ttagtcatcc cttaaattgg ctagcatagg atttttatttt tttctatcct 4020 taatttctat tattaattct gacatgacca ctccagtttc atccctccat gctgagggac 4080 tgatcccatg attgagtaat tcaatgtagt taaaaatatt agtgtatgaa atacctagcc 4140 tctattacac atttttttgg gcgttgccca ctttttttaa cttcctgtaa gtagtaaagc 4200 tcataagtca caagaaatca gattgtttcc ttttgaagga tgagtgacaa atgtgatgcc 4260 ttatggagac tactcttaag gagcagaatt gtattagtct gccatataac acaccttact 4320 tggtcctctt cacctggtcc ttttttttaa atatagctta ttgaggtcag gttccagctt 4380 tgaatttgga tagatgaatt atgaaatggg tataattcta gttttttgtt gcatgtttac 4440 tcaggaacaa acaggaagaa gcaagacaat aatgcaaaag cacaaagtag ctgaactttc 4500 ataggaataa taaagtggtc cttaattgca atgacaacac gttccatgca gagttaaact 4560 tgtacatgct gagtctacaa ctgtaagcct tccttctcca ctctgaggtt ttctagcact 4620 taacacagtc atcctcaaca caaaacatcat ctggggctgc aggaatgtgg atgatactga 4680 cattgtgtgt agcaatctga catatataaa ctattgctga aagtcctcat tagtgttggt 4740 ctaccctcct gtaagaggta gcttaacatg cacattccaa tgtgagtgag ttatgtttct 4800 gtttaggtta acggtgcatg caagtgcaag caatacctaa atctagtgat actagtgctt 4860 ctctgttagt ttttcttttt cttgggaaat ctgaaatggc ttttgaaact gggtaactgt tgattatta ttccacctaa tggaacaagg tagtagcag tgacttctag ccatattctg acattatggc aaacagcata cattatcaa tgttcctgaa gtacccaaaa aagtattgta tttccaatgt aagtgacctt cagctctcct tgaggtgcat actttgtggt caggagaggc 5100 aacatggcat tggttacttg tgaaccattc ctgaatattc cttgtcaagt gtaacaggga ttcccaggaa ctgtggcacg cacttacaag gctgggctgg tgtgtaacta gtgcaggcct aattggcagg attggcttaa atggtcactg gcttgttttg tttcctttt taatcctctt agccatagt tcagatggct tacctacctt tttattgcag aaaaggcagc agaacatgct gatgtagttc cagcagtgag agacactcag gctgcatgtc tatcccaatg aaataggact ttcccttga ctgtatgaca gtggaacacc aaagggattg tttgcttgca ttctaaacta taaagatgg tagttggcta ccgaggtttt tgctgggctg aaattaaggt agtttaacca tctcaggcta cttttgtact tatatgtact tccgtgcctc ttcaag 5566 <210> 2 <211> twenty one <212> DNA <213> Artificial sequence <221> Primer PF <400> 2 ggctagacaa cccctactct c 21 <210> 3 <211> 20 <212> DNA <213> Artificial sequence <221> Primer PR <400> 3 atggccatca ggtggtgatt 20 <210> 4 <211> 193 <212> DNA <213> Artificial sequence <221> PCR amplified fragment containing 27 bp insert <400> 4 ggctagacaa cccctactct ctcccttctt tacaggaaag gtgctccagt agctccagcc 60 ctctgataac attcatggcc cacatctgga cttgctccaa aacatccatg gtgttatgct 120 caggtcccag ggagcagcac aatactctag gtggggtctg agcagagcag ggaaatcacc 180 acctgatggc cat 193 <210> 5 <211> 166 <212> DNA <213> Artificial sequence <221> PCR amplification fragment not containing the 27 bp insert <400> 5 ggctagacaa cccctactct ctcagctcca gccctctgat aacattcatg gcccacatct 60 ggacttgctc caaaacatcc atggtgttat gctcaggtcc cagggagcag cacaatactc 120 taggtggggt ctgagcagag cagggaaatc accacctgat ggccat 166 <210> 6 <211> 27 <212> DNA <213> Artificial sequence <221> 27 bp insert <400> 6 ccttctttac aggaaaggtg ctccagt 27

Claims

1. A chicken growth and development related InDel polymorphic marker, characterized in that: The nucleotide sequence of the InDel polymorphism marker insertion or deletion fragment is shown in SEQ ID NO.6; Chicken with 27 bp insertion YBX3 The sequence of the sixth intron region of the gene is shown in SEQ ID NO. 1, and the insertion site is located at position 934-960 from the 5' end.

2. Use of the InDel polymorphic marker according to claim 1, a primer for detecting the InDel polymorphic marker according to claim 1, and a kit containing the primer for detecting the InDel polymorphic marker according to claim 1 in predicting the growth status of chickens, characterized in that: The following steps are involved: Primers are designed based on the InDel polymorphic markers and PCR amplification is performed. According to the PCR amplification results, the genotype of the tested chicken is judged to be homozygous insertion type II, homozygous deletion type DD or heterozygous type ID, and the growth status of the chicken flock is predicted according to the genotype determination result; the amplified fragments of the primers cover the insertion or deletion fragment region; the growth status includes the growth traits of birth weight, 2-week weight, 4-week weight, 6-week weight, 10-week weight, 12-week weight, 4-week shank circumference, 8-week shank circumference, 4-week chest width, 12-week chest width, 4-week sternum length, 8-week sternum length, 12-week sternum length, 4-week body oblique length, 4-week pelvic width, and 84-day slaughter traits of half-eviscerated weight, full-eviscerated weight, half-eviscerated rate, full-eviscerated rate, head weight, claw weight, wing weight, gizzard weight, pancreas weight, breast muscle weight, leg weight, leg muscle weight, and carcass weight; and the chicken flock is a Gushi chicken×Anka F2 generation resource group.

3. The use according to claim 2, characterized in that: The primers include an upstream primer whose nucleotide sequence is shown as SEQ ID NO.2, and a downstream primer whose nucleotide sequence is shown as SEQ ID NO.

3.

4. Use of the InDel polymorphic marker according to claim 1, a primer for detecting the InDel polymorphic marker according to claim 1, and a kit containing the primer for detecting the InDel polymorphic marker according to claim 1 in assisted selection and molecular breeding of chickens, characterized in that: The assisted selection and molecular breeding are aimed at growth traits and / or slaughter traits; the growth traits are birth weight, 2-week weight, 4-week weight, 6-week weight, 10-week weight, 12-week weight, 4-week shank circumference, 8-week shank circumference, 4-week chest width, 12-week chest width, 4-week sternum length, 8-week sternum length, 12-week sternum length, 4-week body oblique length, and 4-week pelvic width; the slaughter traits are 84-day half-eviscerated weight, full-veined weight, half-eviscerated rate, full-veined rate, head weight, claw weight, wing weight, gizzard weight, pancreas weight, breast muscle weight, leg weight, leg muscle weight, and carcass weight; the chicken flock is a Gushi chicken × Anka F2 generation resource flock.

5. The use according to claim 4, characterized in that Primers are designed based on the InDel polymorphic marker and PCR amplification is performed. According to the PCR amplification results, the genotype of the chicken to be tested is judged to be homozygous insertion type II, homozygous deletion type DD or heterozygous type ID. The growth traits and / or slaughter traits of individuals with genotype II are the best, and homozygous insertion type II individuals are selected for molecular breeding.

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