CN107217094A - One SNP marker related to the gift tilapia speed of growth and its application - Google Patents
One SNP marker related to the gift tilapia speed of growth and its application Download PDFInfo
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Abstract
The invention discloses a gift tilapia SNP marker and its application.Wherein, the SNP marker is SEQ ID NO：The 501st the bit base G or T from holding 5 ' of nucleotide sequence shown in 1.The SNP marker and the speed of growth of gift tilapia of the present invention is closely related, can effective for gift tilapia molecular mark.
Description
Technical field
The present invention relates to SNP marker and its application.In particular it relates to gift tilapia speed of growth correlation
SNP is marked, primer pair and kit for detecting foregoing SNP marker, and foregoing SNP marker, primer pair, kit are lucky rich
Purposes in Tilapia mossambica seed selection, and the method for detecting the gift tilapia speed of growth.
Background technology
Tilapia mossambica belongs to Perciformes, Callichthyidae, Tilapia mossambica category, is eurysalinity tropic fishes, by FAO (Food and Agriculture Organization of the United Nation) FAO
It is classified as one of big Main Foods of the mankind six.At present, Rofe fish culture has spread all over the world more than 80 countries and regions, as worldwide
Cultured fishes.Since nineteen nineties, the good situation of China's Tilapia mossambica industry holding sustainable development, 2016, entirely
The Rofe fish crop of ball is 5,550,000 tons, wherein, Chinese Rofe fish crop is 1,720,000 tons or so, accounts for global total output
31%.However, the seed selection work of China's Tilapia mossambica improved seeds is started late, Tilapia mossambica improved seeds supply wretched insufficiency.
In Java tilapia breeding method, based on traditional sexual control, phenotype breeding technique etc., the effective spe cies identification technology of shortage,
The production of hybrid seeds and conservation mechanism, middle intermolecular hybrid easily, causes to occur in that germplasm degeneration, variet complexity, hybridization in Tilapia mossambica category in addition
The not high phenomenon of generation male ratio, causes Tilapia mossambica seed economic characters to fail, so as to influence the raising of yield and the city of industry
Field competitiveness, restricts the sustainable and healthy development of Tilapia mossambica industry.
Traditional fish breeding method places one's entire reliance upon phenotype, there is cycle length and the low obstacle that can not go beyond of efficiency.
Molecular breeding, i.e. molecular marker assisted selection breeding refers to select breeding material using DNA molecular marker, and synthesis changes
The important economical trait of good seed selection species, is the breeding method that traditional genetic breeding is organically combined with modern molecular biology.Point
Sub- breeding opens up a new way for fish breeding, and with the development of modern biotechnology, molecular labeling is in fish breeding
In effect will become increasingly conspicuous.In the breeding of Tilapia mossambica, it is desirable to by closely related for growth traits, and with number
The selection of the DNA marker of character close linkage is measured, with the target realized early stage seed selection and improve breeding accuracy, so as to obtain more
Big genetic progress.
However, can still have to be excavated effective for the related molecular labeling of the growth traits of Java tilapia breeding at this stage.
The content of the invention
It is contemplated that at least solving one of technical problem present in prior art.Therefore, one object of the present invention
Be to propose it is a kind of related to grouper growth traits, can effective for grouper seed selection SNP marker.
, wherein it is desired to explanation, (single nucleotide polymorphism, SNP, i.e. mononucleotide are more by SNP
State property) it is the molecule that the human genome research center scholar Lander by Massachusetts Institute Technology in 1996 is proposed
Genetic marker, is primarily referred to as in genomic level as the DNA sequence polymorphism caused by the variation of single nucleotide acid.SNP is showed
The polymorphism gone out relates only to the variation of single base, and performance is that have conversion, transversion, insertion and missing etc..
According to an aspect of the present invention, the invention provides a kind of related SNP marker of gift tilapia speed of growth.
Embodiments in accordance with the present invention, the SNP marker is SEQ ID NO：Nucleotide sequence (total length 1001bp) shown in 1 is the from holding 5 '
501 bit bases represent that Y represents G or T with Y.Embodiments in accordance with the present invention, SEQ ID NO：Nucleotide sequence is as follows shown in 1：
ACATGCACGCACACACACACACACACACACACACACACACACACACACAGCTTGGCC
TCACAATTATGTTATGTTACGTGCTGATCTTGGACCTGAGCTGCAGTCCACAAACACAC
ACAGATAACACTAGGTGTGAGGATGAAGTGTTTTCAAACTTAGTCCTTGTGGAGTTAAT
AACCACTACCTGCCTAACTCCTATCTTGATTTTTTTGGCCTCTTGTGTGAAGCAGAACA
AGCTGTGAACTCGACTTTATTATAGCTTAAAAACTACAGTAGCATTCATATGGAGTTGTG
CATCTGGCCACCAGACGAATGTGAAGGCCTTGGTATGTTCAGCTGAAGTTTAAACTTTC
TGATGTCAAATTGGAGGGGGAGCCTTATCCGACAATTTCTGTAACTTTTCAAAATGAAA
GATTCAATTGATCATCTTTTCTAAAGCTCTCTTTTTTCGTCATTTCCACAACTATGGGCAG
AAGCAGTGCAGATAAAAGCAGAAAGYTGTAATATTGAATACATACAGTCTCGAAAAGG
GACTGCGGCAGGAAATGGCCTGCAGCCAGTTTGGCTTGACTTCAGGTCAGTGAGATAC
CTTTACTATATTTCCACATACCATTGTTAGCCTTAAGGTGTGTAGGTTTGCCTCTTTCAAT
CTATTATTTTCTTGACAAAGAGGAAAAACATACTTGATTCTGGAATTATTTCACTTGTTT
CCTCATTTTGCTGAAATTGACCTTTCTGATCCTGTTTCTCTTTGCTTTTTGACACTTCTTG
AGAATTTCAGATGAAGTTCAACTGCAGTGGAAACAAGAGGGGCTTTCCAATTGGCAAG
ATGATTCACTCGCTGTGAACAAAAAAAAAATAGAAGTGGAATTGTAAGACAGTTTTTG
ACTAAGATGGATGCACACACAACAGTTTGTGTCTCAGTAGGATACCGATGTAATCCAAC
ATTTCCTGACTGCAGTTTTTCCCCTGTAATCTATCAAACTGAGGCTGTAGTGT(SEQ ID NO：1).
Inventor has found that genotype is significantly higher than genotype herein for the body weight of heterozygosis GT gift tilapia at the site
For homozygosis TT gift tilapia.And then, embodiments in accordance with the present invention, can by detecting the above-mentioned SNP of gift tilapia
Its speed of growth is effectively determined, specifically, as it was previously stated, the SNP site is notable for the body weight of heterozygosis GT gift tilapia
Higher than the gift tilapia that genotype herein is homozygosis TT, such as when the genotype of the SNP site is GT, then it can determine to be measured
The individual for belonging to fast growth of gift tilapia.Thus, inventor determines, SNP marker of the invention and gift tilapia
The speed of growth be closely related, can effective for gift tilapia molecular mark.And then can be according to reality
Breeding demand growth selection speed carries out Seedling selection to gift tilapia breeding material, is further able to effectively improve breeding
Efficiency and accuracy, improve the genetic level of gift tilapia reproductive population, so as to accurately and efficiently select Ji Fuluo
Non- fish improved seeds.In addition, according to some embodiments of the present invention, gift tilapia point is carried out using the SNP marker of the present invention
Sub- marker-assisted breeding, has the advantages that early screening, saving time, with low cost, accuracy are high.
According to another aspect of the present invention, it is used to detect foregoing SNP of the invention present invention also offers a kind of
The primer pair of mark.Embodiments in accordance with the present invention, the primer pair has SEQ ID NO：Nucleotide sequence shown in 2-3.
Specifically, the sequence of primer pair of the invention is as follows：
Sense primer：5’-CATTCATATGGAGTTGTGCATCT-3’(SEQ ID NO：2)；
Anti-sense primer：5’-GAAAGAGGCAAACCTCACACCTT-3’(SEQ ID NO：3).
Embodiments in accordance with the present invention, can be effectively to the above-mentioned of gift tilapia to be measured using the primer pair of the present invention
Fragment where the SNP marker related to the speed of growth enters performing PCR amplification, and then can effectively be realized to the SNP by sequencing
The detection of mark, determines the genotype in the gift tilapia to be measured SNP marker site, and then can effectively determine Ji Fuluo to be measured
The speed of growth of non-fish.Specifically, genotype is notable for the speed of growth of heterozygosis GT gift tilapia at the SNP marker site
Higher than the gift tilapia that genotype herein is homozygosis TT, such as when the genotype of the SNP site is TT or AT, then it can determine
The individual for belonging to fast growth of gift tilapia to be measured.Thus, for detecting foregoing SNP marks of the invention
Primer pair, can effective for gift tilapia molecular mark, and then can aid in early stage realize the short time,
Low cost, high accuracy ground seed selection gift tilapia improved seeds.
According to another aspect of the invention, present invention also offers a kind of examination for being used to detect foregoing SNP marker
Agent box.Embodiments in accordance with the present invention, the kit is included：It is described previously for the primer of the SNP marker of the detection present invention
It is right.Being included in kit of the invention has SEQ ID NO：The primer pair of nucleotide sequence shown in 2-3.According to the present invention
Embodiment, using the present invention kit included in primer pair, can effectively realize to the upper of gift tilapia to be measured
The polymorphic detection of the SNP marker related to the speed of growth is stated, the gene in the gift tilapia to be measured SNP marker site is determined
Type, and then can effectively determine the speed of growth of gift tilapia to be measured.Specifically, genotype is miscellaneous at the SNP marker site
The speed of growth for closing GT gift tilapia is significantly higher than the gift tilapia that genotype herein is homozygosis TT, such as SNP
When the genotype of point is GT, then it can determine that gift tilapia to be measured belongs to the individual of fast growth.Thus, use of the invention
In the kit for detecting foregoing SNP marker of the invention, it can be aided in effective for the molecular labeling of gift tilapia
Breeding, and then early stage can be aided in realize short time, low cost, high accuracy ground seed selection gift tilapia improved seeds.
In accordance with a further aspect of the present invention, present invention also offers foregoing SNP marker of the invention, primer pair or
Kit, the purposes in gift tilapia seed selection.As it was previously stated, by can be used in detection the present invention and gift tilapia
Reagent the example primer pair or the kit comprising the primer pair etc. as the aforementioned of the related SNP marker of the speed of growth, can be effective
Ground detection determines the genotype of the above-mentioned SNP marker of gift tilapia to be measured, and then the genotype based on acquisition can be effectively true
The speed of growth of fixed gift tilapia to be measured, so as to effectively auxiliary gift tilapia seed selection.
And then, according to another aspect of the present invention, present invention also offers a kind of detection gift tilapia speed of growth
Method.Embodiments in accordance with the present invention, this method passes through the inspection to the foregoing SNP marker of gift tilapia to be measured progress
Survey, determine the speed of growth of the gift tilapia to be measured.Specifically, can be by can be used in the rich with Ji of the detection present invention
Reagent the example primer pair or the kit comprising the primer pair etc. as the aforementioned of the SNP marker of Growth Op Tilapia velocity correlation are right
Gift tilapia to be measured enters performing PCR amplification, sequencing, to detect the gene for the above-mentioned SNP marker for determining gift tilapia to be measured
Type, and then the genotype based on acquisition can effectively determine the speed of growth of gift tilapia to be measured.Wherein, as it was previously stated, should
Genotype is that to be significantly higher than genotype herein be homozygosis TT for the speed of growth of heterozygosis GT gift tilapia at SNP marker sites
Gift tilapia, such as when the genotype of the SNP site is GT, then gift tilapia to be measured belongs to fast growth
Individual.Thus, the method for the detection gift tilapia speed of growth of the invention, can quickly, efficiently and accurately detect Ji Fuluo
The non-fish speed of growth, so can effective for gift tilapia molecular mark, so as to aid in early stage real
Existing short time, low cost, high accuracy ground seed selection gift tilapia improved seeds.
In addition, it is according to the above embodiment of the present invention detection the gift tilapia speed of growth method can also have it is as follows
Additional technical characteristic：
Embodiments in accordance with the present invention, are not particularly limited to the method that gift tilapia to be measured carries out SNP marker detection.
Sequencing, single-strand conformation polymorphism PCR (PCR single strand conformation
Polymorphism, PCR-SSCP), RFLP PCR (PCR-restriTCion
Fragment length polymorphism, PCR-RFLP) and the technology such as flight time mass spectrum can realize SNP inspection
Survey.Wherein, sequencing is that a kind of accuracy highest, flexibility are strong, the detection technique that flux is big, detection cycle is short.Only need to be at SNP
The both sides design pair of primers of point, expands 200-1000bp product, then can directly detect the gene of SNP site by sequencing
Type.Thus, the present invention carries out SNP marker detection using the method for sequencing.According to some specific examples of the present invention, by treating
The detection that gift tilapia carries out foregoing SNP marker is surveyed, the speed of growth of the gift tilapia to be measured is determined, enters one
Step includes：Extract the genomic DNA of gift tilapia to be measured；Using foregoing primer pair, by the lucky rich Rofe to be measured
The genomic DNA of fish enters performing PCR amplification, to obtain pcr amplification product；The pcr amplification product is sequenced, to obtain
Obtain sequencing result；Based on the sequencing result, the genotype of the SNP marker of the gift tilapia to be measured is determined；And
The genotype of the SNP marker based on the gift tilapia to be measured, determines the speed of growth of the gift tilapia to be measured.
Thereby, it is possible to effectively improve the efficiency of the detection gift tilapia speed of growth.
Embodiments in accordance with the present invention, the method for extracting the genomic DNA of gift tilapia to be measured is not particularly limited, can
To be carried out using any of genome DNA extracting method or kit.According to some specific examples of the present invention, using normal
Advise the genomic DNA that phenol chloroform method extracts gift tilapia to be measured.Thereby, it is possible to effectively obtain the base that quality is good, purity is high
Because of a group DNA, it is easy to subsequent step to carry out.
Embodiments in accordance with the present invention, the condition of performing PCR amplification is entered not by the genomic DNA of the gift tilapia to be measured
It is particularly limited.According to some specific examples of the present invention, the amplification system of PCR amplifications is calculated as with 25 μ l：50-100ng/μl
μ l, the 10pmol/ μ l of template DNA 1 SEQ ID NO：The dNTP of upstream and downstream primer each 1 μ l, 10mmol/L shown in 2-3
μ l, the 5U/ μ l of the mix 2.0 μ l of 0.125 μ l, 10 × PCR reaction buffer of Taq archaeal dna polymerases 2.5, surplus is distilled water；Should
PCR amplification reaction condition be：94 DEG C 5 minutes；94 DEG C 30 seconds, 53 DEG C 30 seconds, 72 DEG C 30 seconds, 30 circulation；72 DEG C 5 minutes.
The fragment where SNP marker thereby, it is possible to quickly, efficiently and accurately expand the present invention, obtains target amplification product, is easy to
The progress of subsequent step.
Embodiments in accordance with the present invention, are not particularly limited to the method that the pcr amplification product is sequenced, as long as energy
It is enough effectively to obtain the sequence that pcr amplification product is the fragment where SNP marker.According to some specific examples of the present invention,
It can use at least one selected from HISEQ and single-molecule sequencing method that the pcr amplification product is sequenced.Thus, energy
Enough high fluxs, it is quick, efficiently and accurately obtain sequencing result.
Embodiments in accordance with the present invention,, can be effective by comparing grouper reference gene group sequence based on sequencing result
The genotype for determining the SNP marker of gift tilapia to be measured is TT or GT.
Embodiments in accordance with the present invention, the speed of growth of the GT genotype individuals of the SNP marker is significantly higher than TT or GG
Genotype individuals.Foregoing SNP marker i.e. of the invention and the speed of growth of gift tilapia are closely related.Thus, it is based on
The genotype of the SNP marker of the gift tilapia to be measured determined, can accurately and effectively determine the life of gift tilapia to be measured
Long speed is growth and development traits, such as when the genotype of the SNP site is GT, then grows speed belonging to for gift tilapia to be measured
The fast individual of degree.And then the present invention method can effective for gift tilapia molecular mark, so as to
Auxiliary early stage realizes short time, low cost, high accuracy ground seed selection gift tilapia improved seeds.
It should be noted that the SNP marker related to the gift tilapia speed of growth of the present invention and its application have such as
Lower advantage：
(1) limitation such as age, the sex of the SNP marker that the present invention is provided not by gift tilapia, available for lucky rich Rofe
The early stage seed selection of fish, can remarkably promote the breeding process of gift tilapia；
(2) method of detection gift tilapia the 501st SNP site from holding 5 ' as shown in SEQ ID NO.1, accurately may be used
Lean on, it is easy to operate；
(3) detection of gift tilapia SNP site of the 501st from holding 5 ' as shown in SEQ ID NO.1, is Ji Fuluo
The marker assisted selection of non-fish growth traits provides scientific basis.
The additional aspect and advantage of the present invention will be set forth in part in the description, and will partly become from the following description
Obtain substantially, or recognized by the practice of the present invention.
Embodiment
Unless specifically indicated, term used herein has the general sense in art of the present invention.
Below with reference to specific embodiment, the present invention will be described, it is necessary to which explanation, these embodiments are only explanation
Property, and be not considered as limiting the invention.Unreceipted particular technique or condition in embodiment, according to normal experiment
Condition, such as Sambrook equimoleculars Cloning: A Laboratory Manual (Sambrook J＆Russell DW, Molecular cloning:a
Laboratory manual, 2001), or according to the condition of manufacturer's specification suggestion.Agents useful for same or the unreceipted life of instrument
Manufacturer person is produced, being can be by the conventional products of acquisition purchased in market.
The acquisition of the SNP marker related to the gift tilapia speed of growth of embodiment 1
The acquisition of 1.1 gift tilapia colonies
The Ji Fuluo that the colony used hatches on April 10th, 2016 for Hainan plant of Hua Da ocean science Co., Ltd
Non- fish, on April 20th, 2016,20,000 tail fries are transferred to a net cage and continue to raise.On October 8th, 2016, from the net
Case selects 500 individuals at random, and clip fish body dorsal rags are in -20 DEG C of preservations of 95% ethanol, for extracting genome DNA.
1.2 gift tilapia extracting genome DNAs
This experiment is comprised the following steps that using the genomic DNA in conventional phenol chloroform method extracting gift tilapia fin ray：
(1) take 0.3~0.5g fin rays in 1.5ml Eppendorf pipes, shred, drying of being uncapped on superclean bench
20min；
(2) after ethanol volatilizees substantially, TE buffer solutions (10mmol/ml Tris, 1mmol/ml EDTA, SDS 5%, pH
=8.0) wash 1~2 time, add 600 μ l DNA extracts (0.001mol/L Tris-Cl, 0.1mol/L EDTA, SDS
5%, pH=8.0) and 3 μ l protease k (200mg/ml), 55 DEG C of water-baths digestion 3h or so, the preceding every 10 min jogs centrifugations of 30min
Pipe 1 time, digestion to liquid in pipe clarification；
(3) autogamy phenol chloroformic solution (phenol is added：Chloroform：Isoamyl alcohol=25：24：1) 600 μ l, gently overturn centrifugation back and forth
Pipe 10min, 12000r centrifugation 10min.Take upper strata aqueous phase to be extracted again with isometric above-mentioned phenol chloroform, until aqueous phase and organic phase it
Between there is no white precipitate；
(4) chloroform is used again 1 time, take out supernatant, add the absolute ethyl alcohol precipitation DNA of 2 times of volume precoolings, overturn
Mix, 4 DEG C standing 30min, 12000r centrifugation 10min, precipitation wash again with 70% ethanol, centrifugation dry precipitate after add 50 μ l without
Bacterium water dissolves.4 DEG C save backup or -20 DEG C long-term preservation.
1.3 build simplified gene order-checking (Restriction siteAssociated DNA sequencing, RAD-
Seq) library and it is sequenced and obtains the related SNP marker of gift tilapia body weight
Based on Hiseq2500 high-flux sequence platforms, the gene order-checking method DNA individual to 500 is simplified using RAD
Sample is sequenced, and each individual produces 1G or so data volume, averagely covers 1X gift tilapia genome.It is simultaneously also right
This 500 individuals carry out the growth traits phenotypic evaluations such as body weight.Processing Screening Treatment is carried out to data using PLINK softwares, so
Use the EMMAX softwares based on mixed linear model to carry out GWAS analyses afterwards, one and body are have found from 234,386 SNP
The significantly correlated SNP site of weight.The SNP site is located at shown in SEQ ID NO.1 at the 501bp sites of sequence, in SEQ ID
Site at this is represented in NO.1 sequences with Y, and the base in site is G or T herein.Genotype is heterozygosis GT Ji Fu at the site
The body weight of Tilapia mossambica is significantly higher than the gift tilapia that genotype herein is homozygosis TT.
The sequence verification of the SNP marker related to the gift tilapia speed of growth of embodiment 2 and application
2.1 extract the genomic DNA in gift tilapia fin ray to be measured
Gift tilapia colony of the gift tilapia to be measured in embodiment 1, has randomly selected 200 tail fishes, has pressed again
Genomic DNA is extracted according to the DNA extraction method described in embodiment 1.
2.2 nucleotide fragments of the amplification containing SNP site
Using the foregoing genomic DNA for extracting each gift tilapia to be measured obtained as template, forward primer F is utilized：5’-
CATTCATATGGAGTTGTGCATCT-3’(SEQ ID NO：2) with reverse primer R：5’-
GAAAGAGGCAAACCTCACACCTT-3’(SEQ ID NO：3) nucleotide fragments where SNP to be measured, are amplified.Wherein,
PCR reaction systems are calculated as with 25 μ l：50-100ng/ μ l template DNAs 1 μ l, 10pmol/ μ l primers Fs and R each 1 μ l, 10mmol/L
The μ l of 2.0 0.125 μ l, 10 × PCR reaction buffers of μ l, 5U/ μ l Taq archaeal dna polymerases of dNTP mix 2.5, surplus is double steamings
Water；PCR reaction conditions are：94 DEG C 5 minutes；94 DEG C 30 seconds, 53 DEG C 30 seconds, 72 DEG C 30 seconds, 30 circulation；72 DEG C 5 minutes.
2.3 sequencing identification SNP site genotype
By each pcr amplification product obtained in above-mentioned steps in being unidirectionally sequenced on ABI3730 sequenators, SEQ is recognized
ID NO：The genotype of (i.e. SNP marker of the invention) in 1 sequence at 501bp.Wherein, 200 tails gift tilapia individual to be measured
The genotype and its body weight of the SNP site are as shown in table 1 below.
The genotype and its body weight of 1 200, the table individual SNP site
The association analysis of 2.4 SNP site genotype and the speed of growth
Based on the result of table 1, the genotype that linear analogue analyzes SNP site is carried out using SAS9.0 software Mixed programs
With the relevance of the speed of growth, wherein representing phenotypic number during analysis with whose body weight, the model of use is as follows：
Yijk=μ+Gi+aj+eijk。
Wherein, YijkFor whose body weight value, μ colonies body weight average, GiFor genotype effects vector, ajFor minor-polygene to
Amount, eijkFor random residual effect vector.
The genotype of SNP site and the association analysis result of the speed of growth see the table below 2.
The genotype frequency of the SNP site of table 2 and the association analysis with body weight
As shown in Table 2, the homozygous whose body weight averages of GT heterozygous whose body weight average ratio TT are big.
Association analysis result shown in table 2 shows, average and TT the or GG genotype individuals bodies of GT genotype individuals body weight
The difference of the average of weight reaches pole significance (P<0.01).And then, it was demonstrated that SEQ ID NO：Nucleotide sequence shown in 1 is held from 5 '
The 501st bit base G or T is played, it is significantly correlated with the gift tilapia speed of growth, it is the related SNP of the gift tilapia speed of growth
Mark, the speed of growth of the GT genotype individuals of the SNP marker is significantly higher than TT or GG genotype individuals.
In the description of this specification, reference term " one embodiment ", " some embodiments ", " example ", " specifically show
The description of example " or " some examples " etc. means to combine specific features, structure, material or the spy that the embodiment or example are described
Point is contained at least one embodiment of the present invention or example.In this manual, to the schematic representation of above-mentioned term not
Necessarily refer to identical embodiment or example.Moreover, specific features, structure, material or the feature of description can be any
One or more embodiments or example in combine in an appropriate manner.
Although an embodiment of the present invention has been shown and described, it will be understood by those skilled in the art that：Not
In the case of departing from the principle and objective of the present invention a variety of change, modification, replacement and modification can be carried out to these embodiments, this
The scope of invention is limited by claim and its equivalent.
。
SEQUENCE LISTING
<110>Hainan Hua Da ocean science Co., Ltd
<120>One SNP marker related to the gift tilapia speed of growth and its application
<160> 3
<170> PatentIn version 3.3
<210> 1
<211> 999
<212> DNA
<213> Artificial
<220>
<223> SEQ ID NO：1
<400> 1
acatgcacgc acacacacac acacacacac acacacacac acacacacag cttggcctca 60
caattatgtt atgttacgtg ctgatcttgg acctgagctg cagtccacaa acacacacag 120
ataacactag gtgtgaggat gaagtgtttt caaacttagt ccttgtggag ttaataacca 180
ctacctgcct aactcctatc ttgatttttt tggcctcttg tgtgaagcag aacaagctgt 240
gaactcgact ttattatagc ttaaaaacta cagtagcatt catatggagt tgtgcatctg 300
gccaccagac gaatgtgaag gccttggtat gttcagctga agtttaaact ttctgatgtc 360
aaattggagg gggagcctta tccgacaatt tctgtaactt ttcaaaatga aagattcaat 420
tgatcatctt ttctaaagct ctcttttttc gtcatttcca caactatggg cagaagcagt 480
gcagataaaa gcagaaagyt gtaatattga atacatacag tctcgaaaag ggactgcggc 540
aggaaatggc ctgcagccag tttggcttga cttcaggtca gtgagatacc tttactatat 600
ttccacatac cattgttagc cttaaggtgt gtaggtttgc ctctttcaat ctattatttt 660
cttgacaaag aggaaaaaca tacttgattc tggaattatt tcacttgttt cctcattttg 720
ctgaaattga cctttctgat cctgtttctc tttgcttttt gacacttctt gagaatttca 780
gatgaagttc aactgcagtg gaaacaagag gggctttcca attggcaaga tgattcactc 840
gctgtgaaca aaaaaaaaat agaagtggaa ttgtaagaca gtttttgact aagatggatg 900
cacacacaac agtttgtgtc tcagtaggat accgatgtaa tccaacattt cctgactgca 960
gtttttcccc tgtaatctat caaactgagg ctgtagtgt 999
<210> 2
<211> 23
<212> DNA
<213> Artificial
<220>
<223> SEQ ID NO：2
<400> 2
cattcatatg gagttgtgca tct 23
<210> 3
<211> 23
<212> DNA
<213> Artificial
<220>
<223> SEQ ID NO：3
<400> 3
gaaagaggca aacctcacac ctt 23
Claims (8)
1. a kind of related SNP marker of gift tilapia speed of growth, it is characterised in that the sequence of the SNP marker such as SEQ
ID NO：Shown in 1, the SEQ ID NO：The 501st bit base from holding 5 ' of sequence shown in 1 is G or T.
2. SNP marker according to claim 1, it is characterised in that the growth speed of the GT genotype individuals of the SNP marker
Degree is significantly higher than TT or GG genotype individuals.
3. the primer pair of a kind of SNP marker for described in test right requirement 1 or 2, it is characterised in that the primer pair has
SEQ ID NO：Nucleotide sequence shown in 2-3.
4. the kit of a kind of SNP marker for described in test right requirement 1 or 2, it is characterised in that include：Claim 3
Described primer pair.
5. the primer pair described in SNP marker, claim 3 described in claim 1 or 2 or the kit described in claim 4
Purposes in gift tilapia seed selection.
6. a kind of method for detecting the gift tilapia speed of growth, it is characterised in that by being weighed to gift tilapia to be measured
Profit requires the detection of the SNP marker described in 1 or 2, determines the speed of growth of the gift tilapia to be measured.
7. method according to claim 6, it is characterised in that by carrying out claim 1 or 2 to gift tilapia to be measured
The detection of described SNP marker, determines the speed of growth of the gift tilapia to be measured, further comprises：
Extract the genomic DNA of gift tilapia to be measured；
Using the primer pair described in claim 3, the genomic DNA of the gift tilapia to be measured is entered into performing PCR amplification, so as to
Obtain pcr amplification product；
The pcr amplification product is sequenced, to obtain sequencing result；
Based on the sequencing result, the genotype of the SNP marker of the gift tilapia to be measured is determined；And
The genotype of the SNP marker based on the gift tilapia to be measured, determines the growth of the gift tilapia to be measured
Speed.
8. method according to claim 7, it is characterised in that the speed of growth of the GT genotype individuals of the SNP marker
It is significantly higher than TT or GG genotype individuals.
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| CN108411007A (en) * | 2018-05-25 | 2018-08-17 | 海南晨海水产有限公司 | SNP marker and its application |
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| CN112831575A (en) * | 2021-03-26 | 2021-05-25 | 中国水产科学研究院珠江水产研究所 | Alkaline-resistant SNP marker of Morganbicus mossambica and application thereof |
| CN112877446A (en) * | 2021-03-26 | 2021-06-01 | 中国水产科学研究院珠江水产研究所 | SNP marker related to alkali resistance of tilapia and application thereof |
| CN113005202A (en) * | 2021-03-26 | 2021-06-22 | 中国水产科学研究院珠江水产研究所 | SNP marker related to salt tolerance of tilapia and application thereof |
| CN112877446B (en) * | 2021-03-26 | 2022-06-07 | 中国水产科学研究院珠江水产研究所 | SNP marker related to alkali resistance of tilapia and application thereof |
| CN112831575B (en) * | 2021-03-26 | 2022-06-07 | 中国水产科学研究院珠江水产研究所 | Alkaline-resistant SNP marker of Morganbicus mossambica and application thereof |
| CN113005202B (en) * | 2021-03-26 | 2022-06-07 | 中国水产科学研究院珠江水产研究所 | SNP marker related to salt tolerance of tilapia and application thereof |
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