CN104818331A - Gossypium raimondii functional centromere sequence and molecular marker of same - Google Patents
Gossypium raimondii functional centromere sequence and molecular marker of same Download PDFInfo
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- CN104818331A CN104818331A CN201510225766.4A CN201510225766A CN104818331A CN 104818331 A CN104818331 A CN 104818331A CN 201510225766 A CN201510225766 A CN 201510225766A CN 104818331 A CN104818331 A CN 104818331A
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Abstract
The invention provides a gossypium raimondii functional centromere sequence and a molecular marker of the same. IN the invention, a method of combination of chromatin co-immunoprecipitation and high throughput sequencing (ChiP-Seq) is employed to obtain four DNA sequences in a functional centromere zone of gossypium raimondii, and primers are respectively designed according to the sequences for fluorescence in-situ hybridization (FISH) of mitosis metaphase of the gossypium raimondii. A result proves that all the four sequences generate concentrated, clear and bright signals in the centromere zone of all chromosomes. The centromere sequence can be directly used for positioning the functional centromeres and identification of relative sequences. The molecular marker can be used in genetic location of the centromere of a genetic map of cotton, can provide more accurate information to functional gene location and linkage relationship and can provide basis for sequence splicing of whole genome sequencing of the gossypium raimondii, so that the information of a physical map can be more comprehensive.
Description
Technical field
The present invention relates to information biology and molecular cytogenetics field, be specifically related to the cotton function centromeric sequence of Lei Mengdeshi and molecule marker thereof.
Background technology
Kinetochore is functional element important on higher eucaryote karyomit(e).In cell mitogen with reduction division process, be responsible for chromosomal equalization be separated, thus play the vital role guaranteeing genetic material stable delivery.Because tumor-necrosis factor glycoproteins is rich in region, kinetochore, be difficult to carry out order-checking assembling, therefore, it studies backwardness relatively.Such as, although the genome sequencing of people, fruit bat and Arabidopis thaliana isotype biology completes already, its kinetochore, because of containing a large amount of tumor-necrosis factor glycoproteins, is often still present on gene order-checking collection of illustrative plates with the gap of different lengths, cannot be resolved.Therefore, centric research is no matter for the announcement of its biological characteristics, or research field all important in inhibitings such as gene order-checking.
Along with going deep into of kinetochore research, it is found that, different from the definition of primary constriction on the kinetochore of broad sense and karyomit(e), the region ubiquity of kinetochore functionating a kind of special histone H 3, i.e. CENH3, therefore, kinetochore is truly defined as the Chromatin domains that histone special with kinetochore is combined by researcher, i.e. function centric region.Meanwhile, due to the significance of CENH3 in the definition of kinetochore, it is by the special marking excavated as identification kinetochore functional area and function centromere DNA sequence.Chromatin immune chemical coprecipitation technique (Chromatin immunoprecipitation assay, ChIP) is the main method of DNA and protein interaction in postgraduate's object, and therefore, this method also becomes the effective means that function centromeric sequence is excavated.The method combines with s-generation high throughput sequencing technologies, and (ChIP-Seq) can be separated efficiently in full-length genome level, analytic function centromere DNA sequence, has been widely used at present in the centric research of function.
In recent years, the researchdevelopment of cotton gene group is rapid, but, the centric research of cotton rarely has report, and only report is based on random DNA sequence analysis substantially, screens, cannot determine whether DNA sequence dna is combined with CENH3, therefore its verity as function centromeric sequence remains to be discussed.The present invention utilizes CENH3 antibody to carry out ChIP-Seq experiment, analysis obtains the cotton function centromeric sequence of four Lei Mengdeshi, itself and CENH3 binding relationship clearly, and by the further checking that FISH tests, specify that the location, kinetochore of these sequences and the characteristic of function centromeric sequence thereof., conveniently apply, we develop correspondingly special primer according to its sequence meanwhile, for its clone and application provide strong instrument.
Summary of the invention
The object of the invention is to excavate the cotton function centromeric sequence of Lei Mengdeshi, and develop its corresponding molecule marker, be conducive to the structure of cotton genetic map and physical map, also help kinetochore and form the discussion of exercising mechanism with function, also can provide required sequence information for the structure of cotton artificial chromosome.
For achieving the above object, the present invention adopts following technical scheme:
The cotton function centromeric sequence of Lei Mengdeshi, described sequence obtains according to ChIP-Seq data analysis, title is respectively: sequence 1-Cluster201Contig81, sequence 2-Cluster285Contig8, sequence 3-Cluster291Contig8, sequence 4-Cluster407Contig2, and its nucleotide sequence is respectively shown in SEQ ID NO.1-4.
The corresponding molecule marker primer sequence of described sequence 1-Cluster201Contig81, sequence 2-Cluster285Contig8, sequence 3-Cluster291Contig8 and sequence 4-Cluster407Contig2 is as shown in SEQ ID NO.5-12.
The corresponding molecule marker of the cotton function centromeric sequence of table 1 Lei Mengdeshi
1, the cotton function centromeric sequence of Lei Mengdeshi
The cotton function centromeric sequence of Lei Mengdeshi of the present invention utilizes ChIP-Seq data analysis to obtain.First designed, designed is utilized and available CENH3 antibodies specific ground enrichment CENH3-DNA complex body, then purifying DNA fragment, and through end reparation, add A and after adding sequence measuring joints, reclaim specific size fragment through the pcr amplification of low-circulation number and agarose gel electrophoresis again, complete work prepared by sequencing library.Utilize Illumina Hiseq-2500 sequenator to check order, and by bioinformatic analysis, identify the site that CENH3 is combined with genomic dna, and clearly and the CENH3 DNA sequence dna of combining closely.
2, the design of primer and pcr amplification thereof
The cotton function centromeric sequence of Lei Mengdeshi of the present invention, when designing primer, has following precaution: one is design and have specificity in primer application nucleotide sequence is guarded; Two is that primer length is generally 16-25 bp; Three is that the GC content of primer sequence is generally 40-60%; Four is base stochastic distribution of primer sequence; Five are PCR primer sizes is 200-700 bp.
The condition of pcr amplification is: 95 DEG C of denaturation 3min, 95 DEG C of 30s, 47-53 DEG C of 30s, 72 DEG C of 45s, 35 circulations, 72 DEG C of total elongation 7min.It should be noted that primer sequence is different, annealing temperature is distinguished to some extent: sequence 1 is 50 DEG C, sequence 2 is 47 DEG C, sequence 3 is 53 DEG C, sequence 4 is 51 DEG C.The band of pcr amplification is single and bright, therefore directly adopts QIAGEN test kit to carry out the recovery of PCR primer purifying, its article No.: 28104, title: QIAquick PCR Purification Kit (50).
The invention has the advantages that:
The present invention according to the centric definition of function, designed, designed can the special histone CENH3 in cotton kinetochore carry out ChIP experiment, and the DNA sequence dna by enrichment and after purifying carries out high-flux sequence.This is the effective ways obtaining function centromeric sequence.Pass through bioinformatic analysis, obtain and there is the abundant contigs be combined with CENH3, and tested by FISH, demonstrate the location, kinetochore of sequence, show that the cotton function centromeric sequence of the Lei Mengdeshi of gained of the present invention is true and reliable, efficiently avoid and only carry out from DNA sequence dna level the uncertainty that centromeric sequence seeks.We are according to the corresponding PCR primer of different sequences Design, and primer specificity is good, and the band of amplification is single and bright, therefore is suitable for purified pcr product, label probe, tests for FISH.The clone being designed to function centromeric sequence of PCR primer and application provide important tool.The present invention is that the research of cotton function centromeric sequence have accumulated valuable materials, be conducive to the structure of cotton genetic map and physical map and perfect, also help kinetochore and form the discussion of exercising mechanism with function, also can provide required sequence information for the structure of cotton artificial chromosome.
Accompanying drawing explanation
The cotton Metaphase Chromosomes of results of hybridization-A: Lei Mengdeshi of Fig. 1: Lei Mengdeshi cotton function centromeric sequence, B: kinetochore regional signal, C: composite diagram.As we can see from the figure, the signal that sequence of the present invention produces, all in the region, kinetochore of Lei Mengdeshi cotton colour solid, demonstrates the location, kinetochore of sequence.
Embodiment
The enforcement of embodiment 1:ChIP-Seq technology
ChIP-Seq technology refers to that chromatin immune co-precipitation ChIP combines with new-generation sequencing, and the method is feasible, stable, is not only applicable to the research of individual gene and protein binding relation, is also applicable to the full-length genome research of differential protein binding site.
The basic procedure of the present invention ChIP-Seq used is:
1, the fresh and young tender blade of liquid nitrogen grinding; 2, nucleus is extracted, micrococci ribozyme enzymolysis; 3, add the antibody of the special histone CENH3 in kinetochore, be combined with each other with target protein-DNA mixture; 4, add ProteinA, binding antibody-target protein-DNA mixture, and precipitate; 5, the mixture precipitated is cleaned, remove some non-specific binding; 6, wash-out, obtains the target protein-DNA mixture of enrichment; 7, solution is crosslinked, the DNA segment of purification enrichment; 8, pcr analysis prepare sequencing library; 9, Illumina Hiseq-2500 sequenator is utilized to check order; 10, raw data (reads) is carried out bioinformatic analysis, identify the site that CENH3 is combined with genomic dna, and the DNA sequence dna that clear and definite and CENH3 combines closely.
The concrete operations flow process of ChIP-Seq is:
(1) the fresh and young tender blade of liquid nitrogen grinding, extracts nucleus;
(2) adopt micrococci ribozyme lysed cells core, chromatin is interrupted at random the small segment into 200-1000 bp;
(3) nucleus after enzymolysis is divided into three parts, portion is internal reference (input), and portion adds preimmune serum (Mock), and another part adds the specific antibody mediated immunity precipitating proteins-DNA complex of CENH3, and 4 DEG C of tops turn over night.
(4) after overnight incubation, often pipe adds ProteinA, and 4 DEG C of tops turn 2h;
(5) 4 DEG C of centrifugal 1min of standing 10min, 700rpm, removing supernatant.
(6) low salt solutions, high level salt solution, LiCl and TE washing and precipitating mixture is used successively.Cleaning step is: add solution, and 4 DEG C of tops turn 10min, 4 DEG C of centrifugal 1min of standing 10min, 700rpm, removing supernatant.
(7), after cleaning, wash-out is started.Elutriant is filled a prescription: 1ml solution composition is: 100 μ L10%SDS, 100 μ L1M NaHCO
3, 800 μ LddH
2o.Often pipe adds elutriant, and room temperature top turns 15min, after standing centrifugal, collects supernatant.Repeated washing once.
(8) solution is crosslinked: often pipe adds NaCl, makes its final concentration be 0.2M.Mixing, separates crosslinked spending the night for 65 DEG C.
(9), after separating crosslinked end, often pipe adds RNaseA, hatches 1h for 37 DEG C.
(10) often pipe adds EDTA, Tris-HCl and Proteinase K, 45 DEG C of process 2h.
(11) recovery of DNA fragmentation: adopt QIAGEN glue to reclaim test kit, article No.: 28704, title: QIAquick Gel Extraction Kit (50).
(12) DNA that obtains of purifying through end reparation, add A and after adding sequence measuring joints, then reclaim specific size fragment through the pcr amplification of low-circulation number and agarose gel electrophoresis, prepare sequencing library.
(13) Illumina Hiseq-2500 sequenator is utilized to check order.
(14) utilize the method for information biology, splicing after the reads obtained carries out quality control, obtaining different contigs by checking order; By comparing with reference to genome, removing background interference, obtaining and there is the abundant contigs be combined with CENH3, i.e. function centromeric sequence.In order to confirm the location, kinetochore of this sequence further, the method that we adopt FISH to hybridize, verify the location, kinetochore of these sequences, thus clear and definite its is function centromeric sequence.
According to the centric definition of function, be whether judge DNA sequence dna whether as the core condition of function centromeric sequence with CENH3 protein binding.Therefore, will be from the centric DNA sequence dna of function with CENH3 for the ChIP of means tests the sequence obtained.Because Metaphase Chromosome FISH hybridizes the restriction of resolving power, the so-called centromere DNA sequence of existing cotton, first cannot be clear and definite whether at centric region, secondly also cannot clearly whether bind with CENH3, therefore can not determine whether it is function centromere DNA sequence, the present invention adopts the method for ChIP-Seq, therefore, its sequence is from cotton function centric region, and FISH experiment also demonstrates the validity of the method and the location, kinetochore of sequence.
Embodiment 2: design of primers and pcr amplification
Utilize primer-design software Primer Premier 5.0 pairs of function centromeric sequences to carry out design of primers, impose a condition for: one is total length or the partial sequence of nucleotide sequence; Two are primer lengths is 21bp ± 5 bp; Three be the GC content of primer sequence is 40-60%; Four are PCR primer sizes is 200-700 bp.
The condition of pcr amplification is: 95 DEG C of denaturation 3min, 95 DEG C of 30s, 47-53 DEG C of 30s, 72 DEG C of 45s, 35 circulations, 72 DEG C of total elongation 7min.It should be noted that primer sequence is different, annealing temperature is distinguished to some extent: sequence 1 is 50 DEG C, sequence 2 is 47 DEG C, sequence 3 is 53 DEG C, sequence 4 is 51 DEG C.The band of pcr amplification is single and bright.Therefore directly adopt QIAGEN test kit to carry out the recovery of PCR primer purifying, its article No.: 28104, title: QIAquick PCR Purification Kit (50).
The foregoing is only preferred embodiment of the present invention, all equalizations done according to the present patent application the scope of the claims change and modify, and all should belong to covering scope of the present invention.
SEQUENCE LISTING
<110> University Of Agriculture and Forestry In Fujian
The cotton function centromeric sequence of <120> Lei Mengdeshi and molecule marker thereof
<130> 12
<160> 12
<170> PatentIn version 3.3
<210> 1
<211> 1309
<212> DNA
<213> sequence 1-Cluster201Contig81
<400> 1
aggtcttaaa acgtgattca taagcctcat gaatgtgctt ggggcgtttg ttaagccaaa 60
tggcataacg agtgatacga tccggcccgg gttaatcgag tcgtttcgcg aagttgccca 120
atcgtcgacg aggagtagtt ggagttgtca aaattgggtg atttaggcta agggttgcgg 180
aagctttgag aggggtttag atgagtggtt tggtgttgat aggttcggtt tagaatagaa 240
acaaggtagg ttagaaaaaa ggtaattgat ggagatggaa aatagaactt aaacgtcaag 300
aatgttcaat actaaaaagt attgcccgag tcttatattg cttaaggtga aatttgatga 360
aagatagaag cggaccttga cccgttacct atatggaggt aagaaccaaa ggtataataa 420
tatggttgaa cgccacaccg gttcggtgat aagttcaatg gagttcggat tgacgccact 480
agcaagctag ataagtcgct tcgagactcg aacgaaagaa gagaggaggt aacctcacaa 540
gaacaaagtt ctattaagtt ttaattgatc aaattctgta accttttaca ataaacaagt 600
aagctattta taggcaaaga ctaagctagc cgaatgggca acttaatggc taagaattca 660
gccatgaata tgctagaaat ttctagaaag aaataattaa tttgctgaat gtgcatgaat 720
gaagcattaa aaattcggtt catgtctgga gatgatgcat ggattgaccg aatcatcatg 780
ttgcttcact agatgcattc atgcatcctt agccttgaag aatctccata attccatgaa 840
tgtgcagccc tttaatgatc ctacatctcc cttggctgaa tgaggcatta atgtgcctaa 900
aatacttgaa tggtcatctt gaaaatgcat gaatcatgca tgaaacgtcc catgtacttt 960
cccccataaa tatgatccat gaatcttcaa atacatgaac tttcagcaac tccctcttgc 1020
atgaacgtcc caaatgcatg tgctacttta aatgccatcc attgaacctc aattgtgcat 1080
gcacactccc atacattgca ccaatccgtt catgaacctg cagcaaataa catcagcaag 1140
ttaaatgcat ttcagacaca ttaaattagc agcatatgaa ctcaataatt tgcacattaa 1200
attcggccat acatattaac aaattcagac atatttaaaa catcataatt aattaagtat 1260
ttaatttaga tataattaaa acacttaatt aattatttgt ccagatttt 1309
<210> 2
<211> 1372
<212> DNA
<213> sequence 2-Cluster285Contig8
<400> 2
tgctcttccg atcttagaac ataggcttta taacatgttg catgctgata tgtgacgtat 60
ttaggttcta ctatgtatta tgtgtgtatt taagtatgtt ttggtattga tttgatgttg 120
tttgtgtttt aatgaataag attgcaagtg tttacagctt gtattagtgt agaaactaag 180
ctctataatg tgttacatac tgaagtgtaa catttgcatt agttccattg agtatataag 240
tgtgtttgca ttatgtttac agcttgttaa taagttttca ataagtttgc attactttaa 300
tgcatgtttg ttttgtttac cataaggaaa cgtaatggac agcagcttaa gatatcattt 360
ctagctcggt tgaagctcaa atatgagcat gcagctcatc tcagctcgaa atagttcagt 420
acatttctag ctagctcaaa ctcagctcca ccagctcaac tccagctgac gagtccagaa 480
gcagctgaaa cgtgtcctat gtaagggcaa gtgtcctatg catttgtggc agcccatgag 540
gcatccaaac atgtgaggag cacatgaccg cttgtgcaca agagtgtgtg tgtgcatgtg 600
tagccgaata tacaaggata agaacaaggg aggttgtcgt ttgaaattta aatgtatggc 660
tgttattaaa atgacgccaa aaggagttat aactccttat tttaaaccct tttgattaca 720
gccacgaatt tccattcaac acccctcttt tgtacttata aatacatatt aaaatgatgt 780
attcaggtta gatgatttat gaaaacaatt acgtttgaga atttctctca atttcggttc 840
ttcattgaac tttccttggc atttcagact tagtttgttg tgtcaaattt tcttgatact 900
tcgggttcct agatcgctat ctagagggtc caagtccttt ttgacgttga acagattcga 960
ttcgggtttg ctcagtagat tgtgcaagtt ctttcgagtt taacgttatt ctattccaac 1020
gattactttt gccattgttg aacactcaag ttgctgcctt ggccgtttca tgaagcgttc 1080
gacttcaccg attgaatttc tttttgtttt aaacgccaac aacttcattt tccttccatt 1140
ttcgttcttt tattttaccc atattttacc atgattattt ctttaattcc ttgctgtttt 1200
cttgttgcag gaacgtgctt acaacattcc tagatccgta acatacgaag atagtctgtt 1260
tcgttaggct acatcgtatc atctggtatc cgagcatagg ttggaattgt tgtacttttc 1320
ttctccaaca agaaaattca aaattcaaaa caaatatata tatatatcag tt 1372
<210> 3
<211> 1708
<212> DNA
<213> sequence 3-Cluster291Contig8
<400> 3
tcaaagataa tatcaagagg aggactgaac aatatgtttg agcagccaat aagtcccgca 60
aacgagttgt attcgaaccc ggtgactggg tttggatcca catgcgcaaa gaacgatttc 120
ccgaacaaag gaaatccaaa ttgcaatcaa gaggagacgg cccatttcaa gtgttggagc 180
ggattaacga taacgtttac aagattgacc taccgggtga gtatggagtg agtgcaagtt 240
ttaacgttgc tgatctctct ccttttgatg taggtgacga ttcgaggacg aatcgcttcg 300
aagagaggga ggatgatatg agctcgccca tgaagatagg tgccgaatcc atggagttgc 360
ctttggggcc aattactcga gcacgtgcca agaagttcca agatgctgta gcaagctata 420
ttgctcgatt gtggagtgat gggttgattg cccataaaat gcccagctca actagcttga 480
ttcgcaccat cctacaagct gattttagct cgtgtcagct taattcagct caattcggca 540
cttaattagt tcaaggagct gattatattt attttgaata aatttaatta gttgtgttag 600
ttagaatcag ctcaaaatca tttaattaaa taaatgtgtc ttaatctgga cattcttaat 660
tcagcttatt aaatatgttt tatattagta catgccttta atatgtccat attaagtcat 720
aaattaaatg tgtttagttt aattacaaag ttttaatgtg tcttgactta gacaaattta 780
ttagtggctg ctagttaatt tgtcttagtt aaattaatgt gcagattttt aatatgtcca 840
gctgatgatt tcatgagtat taaacttgtc ttaaccacat ttatttgatg tttttcagct 900
agcttaatgg caaaggaagc tcataaagaa ggagcatgta tttgggcaca tgcatggacg 960
gcatttaaag tgcagctacc ttgtgtttcc agctgatttt ttttgtgtag ctcaatggac 1020
ttcaagctga tattttgagc ttctccaagg gtcaatttcg tggagagcaa tggccaaaag 1080
gtgcttcata aattccagca actaattcac ttggtggcta ctcatattcg gccaaagaag 1140
cataattttt gaagctgtcc atttctcttc tccatagcag caattaagct attttaaaac 1200
attagttgaa tgtgaattat catccttaaa ggggccagcc gaatcatctc ttctagaagg 1260
ttattattca gatttttttt gttgtttatg aatgcatcta ggaaattgct aggaagtttc 1320
ctaaagaagc ttaaggctta agaatctgat tagactattc ggctacctta gtccatccta 1380
taaataggcc atgtaattca cattttgagg ttaatgaaca atttgatagc tttgctaaat 1440
tgtgaggttg ttttcaactc tcgttattct ccaaaaactt gtgagactta tcaaacttct 1500
ttgtggcgtc aaacttgttt ttgtttctac ccgaacttat cactttaatc ccaaaagtgt 1560
ggcgttcgat tcatactaag gttcctatca tctttgatag cgggtcgaag tttctatcca 1620
ctcaaccaac tatccatcca caccttagat cggaagagca cacgtctgaa ctccagtcac 1680
cgtacgtaat ctcgtatgcc gtcttctg 1708
<210> 4
<211> 565
<212> DNA
<213> sequence 4-Cluster407Contig2
<400> 4
aagtacctct tttcgcaaca cttcaagcac cgcacgtaga tgttgaatat gatcttccaa 60
gctcctgcta taaactaaaa tatcatcaaa atatacaaca cagaatttcc caatgaacga 120
acgtaaaaca taattcatga gacgcatgaa agtactagga gcgttggtaa gtccgaatgg 180
cattaccaac cattcataca aaccatactt cgttttgaat gcagttttcc attcgtctcc 240
ctcccgcatt ctgatttgat gatacccact ttttaaatcg atcttcgaaa acaattgggc 300
tccactaagt tcatcaagca tgtcgtcgag gcggggaata ggatggcgat attttattgt 360
gattttgttg atagcgcggc agtcaacgca cattcgccat gatccatcct ttttaggaac 420
caacaatacc ggaaccgcgc acggacttaa gctttcacgg atgtagccct tctccattaa 480
ttcggctact tgcttttcca attccttcgt ttcctcagga ttacttctat atgctggcct 540
atttggaatt gcagcaccgg gcaca 565
<210> 5
<211> 18
<212> DNA
<213> artificial sequence
<400> 5
caaatggcat aacgagtg 18
<210> 6
<211> 18
<212> DNA
<213> artificial sequence
<400> 6
tcaaggtccg cttctatc 18
<210> 7
<211> 18
<212> DNA
<213> artificial sequence
<400> 7
aggataagaa caagggag 18
<210> 8
<211> 16
<212> DNA
<213> artificial sequence
<400> 8
tcaatcggtg aagtcg 16
<210> 9
<211> 20
<212> DNA
<213> artificial sequence
<400> 9
agccaataag tcccgcaaac 20
<210> 10
<211> 23
<212> DNA
<213> artificial sequence
<400> 10
gtcctcgaat cgtcacctac atc 23
<210> 11
<211> 22
<212> DNA
<213> artificial sequence
<400> 11
aatatgatct tccaagctcc tg 22
<210> 12
<211> 18
<212> DNA
<213> artificial sequence
<400> 12
cgaatgtgcg ttgactgc 18
Claims (4)
1. the cotton function centromeric sequence of Lei Mengdeshi, it is characterized in that, described sequence obtains according to ChIP-Seq data analysis, title is respectively: sequence 1-Cluster201Contig81, sequence 2-Cluster285Contig8, sequence 3-Cluster291Contig8, sequence 4-Cluster407Contig2, and its nucleotide sequence is respectively shown in SEQ ID NO.1-4.
2. a molecule marker for the cotton function centromeric sequence of Lei Mengdeshi as claimed in claim 1, is characterized in that: the corresponding molecule marker primer sequence of described sequence 1-Cluster201Contig81, sequence 2-Cluster285Contig8, sequence 3-Cluster291Contig8 and sequence 4-Cluster407Contig2 is as shown in SEQ ID NO.5-12.
3. the cotton function centromeric sequence of Lei Mengdeshi as claimed in claim 1 application in the qualification of the centric location of function and correlated series.
4. the application of molecule marker on the Genes location of cotton genetic map centromere position of the cotton function centromeric sequence of Lei Mengdeshi as claimed in claim 2.
Priority Applications (1)
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105950725A (en) * | 2016-05-13 | 2016-09-21 | 河南师范大学 | Centromere mark realizing fluorescence in situ hybridization with asparagus chromosome and applications of centromere mark |
| CN108315387A (en) * | 2018-02-07 | 2018-07-24 | 北京大学 | Few cells ChIP methods |
| CN113073134A (en) * | 2021-04-06 | 2021-07-06 | 南通大学 | Novel method for carrying out accurate chromosome counting by applying sugarcane centromere specific repeat sequence |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| US20120115132A1 (en) * | 2010-11-05 | 2012-05-10 | Chromatin, Inc. | Identification of centromere sequences using centromere associated proteins and uses thereof |
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| US20120115132A1 (en) * | 2010-11-05 | 2012-05-10 | Chromatin, Inc. | Identification of centromere sequences using centromere associated proteins and uses thereof |
Non-Patent Citations (2)
| Title |
|---|
| SONG LUO: "The Cotton Centromere Contains a Ty3-gypsy-like LTR Retroelement", 《PLOS ONE》 * |
| ZHIYUN GONG: "Repeatless and Repeat-Based Centromeres in Potato: Implications for Centromere Evolution", 《THE PLANT CELL》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105950725A (en) * | 2016-05-13 | 2016-09-21 | 河南师范大学 | Centromere mark realizing fluorescence in situ hybridization with asparagus chromosome and applications of centromere mark |
| CN105950725B (en) * | 2016-05-13 | 2019-07-12 | 河南师范大学 | The centromere of asparagus chromosome fluorescence in-situ hybridization marks and its application |
| CN108315387A (en) * | 2018-02-07 | 2018-07-24 | 北京大学 | Few cells ChIP methods |
| WO2019153852A1 (en) * | 2018-02-07 | 2019-08-15 | 北京大学 | Micro cell chip method |
| CN113073134A (en) * | 2021-04-06 | 2021-07-06 | 南通大学 | Novel method for carrying out accurate chromosome counting by applying sugarcane centromere specific repeat sequence |
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