CN106167825B - A kind of relevant microsatellite marker of yellow catfish growing characteristic and its detection and application - Google Patents
A kind of relevant microsatellite marker of yellow catfish growing characteristic and its detection and application Download PDFInfo
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Abstract
The invention discloses a kind of relevant microsatellite marker of yellow catfish growing characteristic and its detections and application；4 microsatellite locus are found in Pelteobagrus fulvidraco GH sequence, and microsatellite locus is expanded by screening special primer, its relevance with growth traits is analyzed, finds to the advantageous preponderant genotype of growth traits.The result shows that wherein having to growth traits significant or extremely significant related there are three site, site GA and weight, body is long, body is high, body thickness has significant association, has extremely significant association with overall length, caudal peduncle height, and wherein CD is preponderant genotype；Site TCTT with long, caudal peduncle is high significant association, wherein BD is preponderant genotype；Site AC and body be thick, head is with being significantly associated with, wherein DD is preponderant genotype, the 4 microsatellite locus genetic diversity all with higher obtained using primer amplification of the present invention, and having significant or extremely significant relevance with growth traits, the present invention has a very important significance molecular breeding practice.
Description
Technical field
The present invention relates to molecular labeling Biotechnology in Genetic Breeding fields, more particularly, to a kind of yellow catfish growing characteristic phase
The microsatellite marker of pass and its detection and application.
Background technique
Pelteobagrus fulvidraco is the small-sized economic freshwater fish that China cultivates extensively；Because of its fine and tender taste, full of nutrition, dressed fish is high
Very popular, Pelteobagrus fulvidraco breeding parent is mainly wild fishing or the population for cultivating number generation, does not pass through artificially breeding,
Its speed of growth, disease resistance and culture performance are weaker, therefore the two kinds of breedings work for carrying out Pelteobagrus fulvidraco has actual meaning.
A large amount of research has been carried out in terms of Pelteobagrus fulvidraco genetic breeding there are many scholar at present, and achieve much at
Fruit.With the development of biotechnology, genetics-breeding in fish technology has more and more methods, and molecular labeling is primarily referred to as can
Reflect the genetic fragment of certain othernesses in genome between individual biology or group, high, the widely distributed, codominance with polymorphism
Etc. a series of advantages, genetics-breeding in fish and genetic linkage maps building, Relationship iden- tification, in terms of
It has a wide range of applications.
Microsatellite marker (microsatellite), also known as short tandem repeat (short tandem repeats,
It STRs) or simple repeated sequence (SSRs), is the simple repeated sequence being uniformly distributed in eukaryotic gene group, by 2~6
The tandem repeat of nucleotide is constituted, since the number of repetition of recurring unit is in variability and quantity is rich between individuals
It is rich.Each microsatellite DNA is made of core sequence and flanking sequence, and core sequence is arranged in tandem sequence repeats, flanking DNA
Sequence is located at the both ends of core sequence, for conservative special single-copy sequence, microsatellite can be made specifically to be positioned at chromosome normal
The privileged site in chromatin area.
Microsatellite marker is the second generation molecular labeling based on PCR, and so far, micro-satellite labeling technique is important
Economic fish has been widely used, and becomes research population genetic diversity, population genetic variations, genetic map construction, function
Can the assignment of genes gene mapping and QTL analysis etc. an important technology.The excellent high-yield variety of growth traits is cultivated using molecular labeling
It is one of the important channel increased economic efficiency, height can be assisted by obtaining gene relevant to yellow catfish growing characteristic and gene loci
The breeding of Pelteobagrus fulvidraco is produced, there is presently no gene relevant to yellow catfish growing characteristic and the reports of gene loci.
Summary of the invention
The technical problem to be solved by the present invention is to overcome drawbacks described above of the existing technology, a kind of and Pelteobagrus fulvidraco is provided
The relevant microsatellite marker of growth characteristics.
A second object of the present invention is to provide the primers for the above-mentioned microsatellite marker of specific detection.
Third object of the present invention is to provide the applications of above-mentioned microsatellite marker.
The purpose of the present invention is what is be achieved by the following technical programs:
It is a kind of for expanding the primer of microsatellite relevant to yellow catfish growing characteristic, the nucleotide sequence of primer such as SEQ
Shown in NO:4~9 ID, wherein the repetition motif of microsatellite segment of primer SEQ ID NO:4 and SEQ ID NO:5 amplification is
(GA)n1, the value range of n1 is 17~35, the weight of the microsatellite segment and Pelteobagrus fulvidraco, overall length, body are long, body is high, body is thick,
Caudal peduncle is high related；The repetition motif of the microsatellite segment of primer SEQ ID NO:6 and SEQ ID NO:7 amplification is (TCTT)n2, n2
Value range be 6~14, the microsatellite segment and Pelteobagrus fulvidraco it is long, caudal peduncle is high related；Primer SEQ ID NO:8 and
The repetition motif of the microsatellite segment of SEQ ID NO:9 amplification is (AC)n3, the value range of n3 is 25~35, the microsatellite
The body of segment and Pelteobagrus fulvidraco is thick, long related.
Preferably, the solution temperature of the microsatellite segment of primer SEQ ID NO:4 and SEQ ID NO:5 amplification is 56 DEG C,
Clip size is 311bp；The solution temperature of the microsatellite segment of primer SEQ ID NO:6 and SEQ ID NO:7 amplification is 61 DEG C,
Clip size is 272bp, and the solution temperature of the microsatellite segment of primer SEQ ID NO:8 and SEQ ID NO:9 amplification is 56 DEG C,
Clip size is 220bp.
Applicant passes through the study found that with above-mentioned micro-satellite primers amplified sample, specifically can amplify microsatellite marker
The growth form of (or being called microsatellite locus), these microsatellite markers and Pelteobagrus fulvidraco is closely related, therefore can be used for yellow forehead
The molecular breeding of fish.
The present invention also provides the kits for containing the micro-satellite primers.
According to embodiments of the present invention, the complete of Pelteobagrus fulvidraco can effectively can be determined by detecting above-mentioned microsatellite marker
Long character, therefore, the present invention also provides the application of the micro-satellite primers or the kit in Pelteobagrus fulvidraco genetic breeding.
The present invention also provides a kind of methods for detecting Pelteobagrus fulvidraco overall length character, comprising the following steps:
S1. sample DNA is extracted；
S2. the primer amplification sample DNA for utilizing as claimed in claim 1 or 22 NO:4~9 the SEQ ID, is analyzed in sample DNA
The type and genotype of microsatellite marker determine the overall length character of Pelteobagrus fulvidraco.
The present invention also provides a kind of methods of the excellent Pelteobagrus fulvidraco of breeding, comprising the following steps:
S1. sample DNA is extracted；
S2. the primer amplification sample DNA for utilizing as claimed in claim 1 or 22 NO:4~9 the SEQ ID, is analyzed in sample DNA
The type and genotype of microsatellite marker；
S3. result judges: if motif (GA) containing repetition in sample DNAn1, then selecting genotype for the sample of CD is excellent
Pelteobagrus fulvidraco；If motif (TCTT) containing recombination in sample DNAn2, then selecting genotype for the sample of BD is excellent Pelteobagrus fulvidraco；If sample
Motif (AC) containing repetition in DNAn3, then selecting genotype for the sample of DD is excellent Pelteobagrus fulvidraco.
Compared with prior art, the invention has the following advantages:
The present invention obtains Pelteobagrus fulvidraco GH full length gene sequence for the first time, finds 4 in GH sequence by SSRHunter software
A microsatellite locus analyzes the relevance of Pelteobagrus fulvidraco microsatellite locus and growth traits by SAS software GLM model for the first time, warp
Variance analysis test is in the site of significant difference, is carried out using genotype of the Duncan method to significant difference site multiple
Compare, finds to the advantageous preponderant genotype of growth traits.Inspection result are as follows: wherein there are three sites and growth traits to have significantly
Or extremely significant correlation, site GA and weight, body are long, body is high, body thickness has significant association, there is extremely significant association with overall length, caudal peduncle height,
Wherein CD is preponderant genotype；Site TCTT with long, caudal peduncle is high significant association, wherein BD is preponderant genotype；Site AC
With body thickness, head with (P < 0.05) is significantly associated with, wherein DD is preponderant genotype, the microsatellite locus and growth of this test
Shape has the probability of significant relevance bigger, this may be located on GH gene due to newfound 4 microsatellites, and polymorphism is to Huang
The growth and development of forehead fish has direct influence.The study find that 4 microsatellite locus genetic diversity all with higher,
And site GA, TCTT, AC have significant or extremely significant relevance with growth traits, the present invention, which practices molecular breeding, to be had
It is of great significance.
Detailed description of the invention
Fig. 1 is common amplification PCR result, wherein Fig. 1 (1) is site AT, GA primer electrophoresis result；Fig. 1 (2) is site
TCTT, AC primer electrophoresis result；Most left band be MarkerDL2000, from top to bottom stripe size difference 2000bp, 1000bp,
750bp、500bp、250bp、100bp。
Fig. 2 is genotyping result of the microsatellite molecular marker PCR product on ABI3730xl；Wherein, Fig. 1 (1) is the site AT
Parting；It 1(2) is the site GA parting；It 1(3) is the site TCTT parting；It 1(4) is AC Locus Analysis in Shoots.
Specific embodiment
The contents of the present invention are further illustrated below in conjunction with Figure of description and specific embodiment, but should not be construed as pair
Limitation of the invention.In the case where without departing substantially from spirit of that invention and essence, modify made by the method for the present invention, step, condition
Or replacement, it all belongs to the scope of the present invention.Unless otherwise noted, experimental method used in embodiment is those skilled in the art
Conventional method and technology known to member, reagent or material are to be obtained by commercial sources.
The present invention obtains Pelteobagrus fulvidraco GH overall length (sequence is as shown in SEQ ID NO:1) using the method for homologous clone, altogether
3117bp, 5 exons (being referred to as exons 1, exon 2, exon 3, exon 4 and exon 5 since 5 ' ends),
4 intrones (are referred to as introne 1, introne 2, introne 3, introne 4 since 5 ' ends), and 5 ' UTR are 165bp, and 3 '
UTR is 485bp.
The screening and parting of 1 microsatellite locus of embodiment
Using the microsatellite locus in SSRHunter software search Pelteobagrus fulvidraco GH full length gene sequence, microsatellite position is determined
The information such as vertex type, position and length.Using 5.0 software of Primer Premier in the site SSRs upstream and downstream 150bp
Left and right design upstream and downstream primer.Primer information is shown in Table 1.
The semi-automatic detection of fluorescence in order to realize microsatellite polymorphism needs to carry out the 5 ' of general primer or 3 ' ends glimmering
The label of light modification group.The PCR product that fluorescent primer expands carries out on 96 hole capillary 3730xl DNA analysis instrument
Electrophoresis detection, fluorophor can issue coloured light under the effect of the laser, and system collects the fluorescence letter of different colours automatically
Breath, the final data information for obtaining PCR product.
Regular-PCR amplified reaction is carried out using the primer in table 1, to grope the optimal reaction condition of primer, reaction system
For table 2.
PCR response procedures: 94 DEG C of 5 min of initial denaturation；38 circulations (are denaturalized 94 DEG C of 30 s, Tm draws according to different
Object is different, and anneal 51~56 DEG C of 30s, extends 72 DEG C of 25s)；Extend 72 DEG C of 10 min, 4 DEG C of forever afterwards.
Product carries out 2% agargel electrophoresis detection, is taken pictures and is saved using gel imaging system after electrophoresis；Whole process is protected from light
Operation, the PCR product that will amplify purpose band is wrapped with masking foil send Huada gene company to carry out parting detection.
3 kinds of fluorescent decoration groups of this experimental selection, the site AT and AC select FAM(green, green), the site GA be HEX
(yellow, yellow), the site TCTT are TAMRA(yellow-orange, crocus).
The result shows that: 4 microsatellites are detected on GH gene, site is distributed in introne 1, introne 3,3 ' UTR, all
Belong to simple SSR, core sequence is respectively (AT) n, (GA) n1, (TCTT) n2, (AC) n3.Be respectively designated as AT, GA, TCTT,
AC.Before fluorescent primer synthesis, PCR amplification first is carried out to examine the quality of primer amplification with general primer, is then synthesized again
Fluorescent primer, and mark fluorescent groups, as seen in Figure 1 each site SSRs energy are held in 5 ' in upstream (or downstream) primer
It enough finds the respective value of multiple annealing temperatures and proves that this 4 sites SSRs can be amplified.
Determined using the genotype that microsatellite PCR product application 96- capillary 3730xl DNA analysis instrument carries out SSRs, tool
Body step are as follows:
(1) it dilutes the pcr amplification product of SSR: taking one piece of 96 hole PCR plate, 10ul ddH is added in every hole2O, then every hole adds
Entering sample 1ul(multitube to mix is also so operation).Dilution on-fixed, depending on the concentration of each SSR amplified production.Slightly from
The heart mixes.
(2) it prepares internal standard ROX500 mixed liquor: taking a 1.5ml centrifuge tube, addition is changed to 1020ul Hi-Di
Formamide and 60ul ROX500, vortex oscillation mixing (the amount of reagent ratio of each reaction is Hi-Di Formamide:
ROX500=8.5ul:0.5ul).
(3) one piece of 96 new hole PCR plate is separately taken, every hole is added sample and dilutes mixed liquor 1ul, and then every hole is separately added into again
9ul ROX500 mixed liquor, slightly centrifugation mix.
(4) denaturing samples, 95 DEG C 4 minutes, 4 DEG C 1 minute.
(5) sequenator 3730xl electrophoresis on.
(6) application software GeneMapper ID v3.1 software is to the reproducibility of electrophoresis, the analysis result of segment and base
Because genotyping result is checked, finally output is as a result, such as Fig. 2.
Wherein the small peak beside main peak is shadow peak, in gel electrophoresis then be known as shadow band/ghost band, be because
Caused by the sliding mispairing of Taq enzyme.STRs genotyping result from top to bottom are as follows:
AT: fluorescent marker FAM, it is homozygote that molecular size range, which is 252,；
GA: fluorescent marker HEX, molecular size range 321,342 is heterozygote；
TCTT: fluorescent marker TAMRA, it is homozygote that molecular size range, which is 286,；
AC: fluorescent marker FAM, molecular size range 216,230 is heterozygote.
2 SSRs allele of embodiment and distribution
Totally 160 tail, site AT detect 3 allele, site TCTT and site AC detection to the present embodiment experiment sample
To 4 allele, site GA detects 5 allele.The allele in each site and distribution are shown in Table 3.
The analysis of 3 SSRs heredity growth characteristics of embodiment
The Pelteobagrus fulvidraco Population genetics analysis of 4 SSR polymorphic sites is shown in Table 4 in research.The average heterozygosity in 4 sites
It is 0.5661, site AT and TCTT are moderate polymorphic, and site GA and AC are that height is polymorphic, illustrate the polymorphism of this 4 microsatellites
It is higher.
This research carries out parting to SSRs in 96- capillary ABI3730xl, utilizes the general linear model in SAS software
(GLM), to microsatellite locus genotype and the main growth traits of Pelteobagrus fulvidraco, (weight, overall length, body are long, body is high, body is thick, long, tail
Handle is long, caudal peduncle is high) it is associated analysis, Analysis of variance examines the site in significant difference, is carried out using Duncan method more
Compare again.The result shows that: the growth traits in the AT of site between each genotype is without significant difference (table 5).In the GA of site, genotype
BE and CD weight is significant to overweight genotype AB (P < 0. 05)；Genotype BE, DD, EE overall length is considerably longer than (P < 0. genotype AB
05), and genotype CD overall length is extremely significant is longer than genotype AB (P < 0. 01)；Genotype BE, CD, EE body is long to be considerably longer than gene
Type AB (P < 0. 05)；Genotype BE, CD body height is significantly higher than genotype AB (P < 0. 05)；Genotype CD body thickness is significantly thicker than
Genotype AB and BB (P < 0. 05)；Genotype BB, BC, BD, CD, DD, EE caudal peduncle height is significantly higher than genotype AB (P < 0. 05),
And genotype BE caudal peduncle is high extremely significant higher than genotype AB (P < 0. 01) (table 6 and table 7).
It is genotype CC long to be considerably longer than genotype BB (P < 0. 05) in the TCTT of site；Genotype BD caudal peduncle height is significantly high
In genotype AB (P < 0. 05) (table 8 and table 9).In the AC of site, genotype CC and DD body thickness is significantly thicker than frequency of genotypes AA (P < 0.
05)；It is genotype BB, CC, CD, DD long to be considerably longer than genotype BD (P < 0. 05) (table 10 and 11).
Note: it is significant (P < 0. 05) with a line difference lowercase letter indication difference, difference is indicated with a line difference capitalization
Extremely significant (P < 0. 01).
4, the site SSRs is found altogether on Pelteobagrus fulvidraco GH gene, research shows that site AT and growth traits onrelevant；Site
GA has with weight, body length, body height, body thickness is significantly associated with (P < 0.05), has extremely significant association (P < with overall length, caudal peduncle height
0.01) eight kinds of genotype, are shared, wherein CD is preponderant genotype；Site TCTT and long, caudal peduncle be high to be had and is significantly associated with (P <
0.05) 7 kinds of genotype, are shared, wherein BD is preponderant genotype；Site AC and body be thick, head is with being significantly associated with (P < 0.05),
6 kinds of genotype are shared, wherein DD is preponderant genotype.
Comparative example 1
When designing micro-satellite primers, each pair of primer all devises two pairs, naming method are as follows: site-F1/R1, totally four
Microsatellite locus AT, GA, TCTT, AC.Red is labeled as finally expanding PCR primer, and carries out fluorescent decoration.
When inventor is according to 4 microsatellite design primers, many primer pairs are devised, primer given below cannot be special
Specific amplification microsatellite locus can not be used for Genotyping.
SEQ ID NO:10(AT-F1): 5'-GCTCTGTTACCTGATGAA-3'
SEQ ID NO:11(AT-R1): 5'-TGAGTATTGAAGCACTTT-3'
SEQ ID NO:12(GA-F2): 5'-GAGGGAAACCTGAGGAAGA-3'
SEQ ID NO:13(GA-R2): 5'-GGTGAATGGAGGAGAATAA-3'
SEQ ID NO:14(TCTT-F2): 5'-GATCCCTGGATTCCAACT-3'
SEQ ID NO:15(TCTT-R2): 5'-AGCACCATCGTGAAACTG-3'
SEQ ID NO:16(AC-F1): 5'-AAAATCCACCACTAGACC-3'
SEQ ID NO:17(AC-R1): 5'-ACTCACCAACACCACAGA-3'.
SEQUENCE LISTING
<110>Agricultural University Of South China
<120>a kind of relevant microsatellite marker of yellow catfish growing characteristic and its detection and application
<130>
<160> 17
<170> PatentIn version 3.3
<210> 1
<211> 3117
<212> DNA
<213>GH gene order
<400> 1
tgatgttatt gtggaaggtg tgaggtgtct catgtcgatg cattaagcat gtagagatca 60
ggtataaaaa ttggacctgc tagcaggagg gaaatcattg tctgaaagtc ggcgtctgat 120
ttttcagtga aaattcgaac caagtttctt cagagagatt ctaaaatggc tcgaggtaag 180
gctctgacct ggtgttacat tacgactgtg atgttacagt gaggaacttt tattgaygtt 240
tttgttttat aagattacta ttagattttc aaattacttt ctgtataaaa tccaccacta 300
gacctgtttt attttgtgcc taatttcaga attatccttg tagatgtatc ctcacagtac 360
acacacacac acacacacac acacacacac acacacacac acacacacac acacacacat 420
actgtatatg cattatataa tgtatgctac tctcctgctg ttcttcctct tccacagggt 480
tagtgttgct ctctgtggtg ttggtgagtt tgttctttag ccaaggcgcg acatttgaga 540
accagaggct cttcaacaac gccgtcattc gtgtgcaaca ccttcatcag ctggctgcca 600
agatgatgga tgactttgta agctgacttt acatcattta cagatgttta gtttactgaa 660
gtatgcttta gctaacatgg ttgctgtgct gcaaatataa aggaggaagc tctgttacct 720
gatgaacgca aacagctcag caagatcttc cccctgagtt tctgcaactc ggactccatc 780
gaagctccgg caggcaaaga cgagacccag aaaagctctg tgagtcacaa aactgcctca 840
agaattcatg cgattttaat atatatatat ttaaagatat ttgttattgt gataacgaat 900
ttggtgattt aatgatcatt agctgtgaca caaatgattt aaaggagcaa tatgtaatat 960
ataaaagtgc ttcaatactc ataaaaaatt gaatcatgtc aaagatgctt taaaatgttc 1020
aaaaaaatgt aataaagtag ttgttttact gtttttacag tttctttgat taaggcttct 1080
acttaattaa ttacccaata aaatatcccc cccccaatat ttaaatgttt cttaaaagaa 1140
ataactaaaa aacattactt tttctgaatt ttttaagagt agccctgcaa tgcaaggtat 1200
ctattttaaa gtttcagtta acatttttct ggcccagaaa caagctccac tgtttttttt 1260
tttttttgta aaaggcagct taagagtcaa acatgtctag ctagtagttg atgttttcat 1320
tgctctctta gcgtgatgaa tattgtgatt tacgtgctac agtctcatgc taaagatcgt 1380
ttataagcaa acatgagcat cggagagaag aatactgctt tgcatctctg tgaactttaa 1440
acgagtctgt tttttaacca ggtgctgaag ctactgcaca cttcctaccg tctgatcgag 1500
tcatgggagt tccccagcaa gaacctcggc aaccctaacc atatctctga gaagctggct 1560
gacctgaaga tgggcatcgg cgtgcttatc gaggtgagca cgagcgccac agatcagaca 1620
ccgagtcact gcaatgtgtg cagggagaac agaagagtta atagaatgtg ttagttaaca 1680
tgtatgagta tgtatttccc gtatggacct gtgtgcacaa aaatgatgtt ttgtacaact 1740
tggatgaaag tgtttttttt gtgtcattcc ccatccattt ctctataact aatacatgct 1800
ctggtgctct gggtggctca gaacttgcga tctaagtaca agacttagca gtacaagaaa 1860
cccgtcagtt cattcagcta tatgcggttt taaaacttaa aactaatcag aatagtagct 1920
atttatttac tgttggtaga atctgaccgt tcattcattc aatggaaaag ttggtaaggt 1980
aataataatg taacttttgt aactgtaagt ttgcagttct gatcaattga aataatttta 2040
atcattaaaa tgtaaatgtt tacaccatat gttgtagtct caggaggata aactcttcat 2100
accttccagc cctattaacg tccccggctc agacccggag ctctgctcct ttacacatga 2160
gtgaaattta aacagtaaca attattagta ttatttagat cattttctcc agtattgtag 2220
tttttatata gcgcaggttg gatttattta actcctacat gttaataaac acaataacca 2280
acaggcgtac ttaaggtggt cgttatttat gatcaattca tgtattgtta ttataaagtg 2340
tttcagttat gcagtatgat aaagtttagt gaaaattgtc tagttttatt gtagcaaggc 2400
ttataaagtc actctgctca ctccataggg atgtctggac ggacagacca gcatggacga 2460
gaacgacgct ctggctccgc cattcgagga tttctaccag accttgagcg agggaaacct 2520
gaggaagagc ttccgtctgc tgtcctgctt caagaaggac atgcacaagg tggagaccta 2580
tctaagcgtg gccaagtgca ggagatccct ggattccaac tgcaccctgt agaatgagag 2640
agagagagag agagagagag agagagagag attgattgat tgattgacta gtcacagtct 2700
gggatctctc caggcatgac tatcatcttt ctttctttct ttctttcctt ttgtttcttt 2760
gggtttaatt tctgtcttta tgtatttatt ctcctccatt cacctgttta acagatttct 2820
ccatctcagc aaccttaagg atgtaacata tgggaccagt ttcacgatgg tgctatgcaa 2880
attcagcttt atatcaaata acgaataaga aaatggatta aacgtaatta atttagaaat 2940
ggacgaggac gctcctggct aatgaatgta aaatattcag agggtatatt ttcctatcaa 3000
agtctatact cttctaattt taattatggc ctatttctgt gtgaagcttt gactgtctct 3060
gtgtgaaaat ctctctcatt gcttttttgt ggatttccaa taaacctaat ccctggg 3117
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<212> DNA
<213> AT-F
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gtttctgcaa ctcggact 18
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<213> AT-R
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gcatctttga catgattc 18
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<213> GA-F
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ctaccagacc ttgagcga 18
<210> 5
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<212> DNA
<213> GA-R
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ggtgaatgga ggagaata 18
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<211> 19
<212> DNA
<213> TCTT-F
<400> 6
gatccctgga ttccaactg 19
<210> 7
<211> 19
<212> DNA
<213> TCTT-R
<400> 7
tagcaccatc gtgaaactg 19
<210> 8
<211> 17
<212> DNA
<213> AC-F
<400> 8
aatccaccac tagacct 17
<210> 9
<211> 17
<212> DNA
<213> AC-R
<400> 9
ctcaccaaca ccacaga 17
<210> 10
<211> 18
<212> DNA
<213> AT-F1
<400> 10
gctctgttac ctgatgaa 18
<210> 11
<211> 18
<212> DNA
<213> AT-R1
<400> 11
tgagtattga agcacttt 18
<210> 12
<211> 19
<212> DNA
<213> GA-F2
<400> 12
gagggaaacc tgaggaaga 19
<210> 13
<211> 19
<212> DNA
<213> GA-R2
<400> 13
ggtgaatgga ggagaataa 19
<210> 14
<211> 18
<212> DNA
<213> TCTT-F2
<400> 14
gatccctgga ttccaact 18
<210> 15
<211> 18
<212> DNA
<213> TCTT-R2
<400> 15
agcaccatcg tgaaactg 18
<210> 16
<211> 18
<212> DNA
<213> AC-F1
<400> 16
aaaatccacc actagacc 18
<210> 17
<211> 18
<212> DNA
<213> AC-R1
<400> 17
actcaccaac accacaga 18
Claims (6)
1. a kind of for expanding the primer of microsatellite relevant to yellow catfish growing characteristic, which is characterized in that the nucleotide of primer
Sequence is as shown in NO:4~9 SEQ ID.
2. primer according to claim 1, which is characterized in that primer SEQ ID NO:4 and SEQ ID NO:5 is expanded micro-
The repetition motif of satellite segment is (GA)n1, the value range of n1 is 17~35, the weight of the microsatellite segment and Pelteobagrus fulvidraco,
Overall length, body are long, body is high, body is thick, caudal peduncle is high related；The microsatellite segment of primer SEQ ID NO:6 and SEQ ID NO:7 amplification
Repeating motif is (TCTT)n2, the value range of n2 is 6~14, long, the high phase of caudal peduncle of the microsatellite segment and Pelteobagrus fulvidraco
It closes；The repetition motif of the microsatellite segment of primer SEQ ID NO:8 and SEQ ID NO:9 amplification is (AC)n3, the value range of n3
It is 25~35, the body of the microsatellite segment and Pelteobagrus fulvidraco is thick, long related.
3. primer according to claim 1, which is characterized in that primer SEQ ID NO:4 and SEQ ID NO:5 is expanded micro-
The solution temperature of satellite segment is 56 DEG C, clip size 311bp；Primer SEQ ID NO:6 and SEQ ID NO:7 is expanded micro-
The solution temperature of satellite segment is 61 DEG C, clip size 272bp, and primer SEQ ID NO:8 and SEQ ID NO:9 is expanded micro-
The solution temperature of satellite segment is 56 DEG C, clip size 220bp.
4. the kit containing any primer of claim 1-3.
5. application of the kit in Pelteobagrus fulvidraco genetic breeding described in any primer of claim 1-3 or claim 4.
6. a kind of method for detecting Pelteobagrus fulvidraco overall length character, which comprises the following steps:
S1. sample DNA is extracted；
S2. the primer amplification sample DNA for utilizing any NO:4~9 the SEQ ID claim 1-3, is analyzed in sample DNA
The type and genotype of microsatellite marker determine the overall length character of Pelteobagrus fulvidraco.
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| CN107155959B (en) * | 2017-04-28 | 2020-08-11 | 南京师范大学 | Compound breeding method for improving growth traits of pelteobagrus fulvidraco |
| CN108432671B (en) * | 2018-03-27 | 2021-06-01 | 南京师范大学 | Method for creating new strain of pelteobagrus fulvidraco by backcross breeding |
| CN111154896A (en) * | 2020-03-14 | 2020-05-15 | 甘肃省水产研究所 | SNP marker related to Gansu golden trout growth traits and method application thereof |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104357553A (en) * | 2014-10-14 | 2015-02-18 | 华中农业大学 | Pelteobagrus fulvidraco microsatellite family identification method |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104357553A (en) * | 2014-10-14 | 2015-02-18 | 华中农业大学 | Pelteobagrus fulvidraco microsatellite family identification method |
Non-Patent Citations (3)
| Title |
|---|
| 甘肃金鳟生长激素基因多态性与生长性状的相关分析;虎永彪等;《淡水渔业》;20140930;第44卷(第5期);100-103 |
| 黄颡鱼微卫星标记的开发及其遗传连锁图谱的构建;张秀杰;《华中农业大学博士学位论文》;20111231;第14，25，35-36，39页 |
| 黄颡鱼微卫星标记筛选及特征分析;刘臻等;《农业生物技术学报》;20080815(第04期);604-609 |
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