CN104818331B - Lei Mengdeshi cotton function centromeric sequences and its molecular labeling - Google Patents
Lei Mengdeshi cotton function centromeric sequences and its molecular labeling Download PDFInfo
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Abstract
The invention provides Lei Mengdeshi cotton function centromeric sequences and its corresponding molecular labeling.We are combined using chromatin immune co-precipitation with high throughput sequencing technologies（ChIP‑Seq）Method obtained four DNA sequence dnas in Lei Mengdeshi cotton function centromeres region.Primer is separately designed according to the sequence, for Lei Mengdeshi cotton mitosis metaphase FISHs（FISH）.As a result show, this four sequences are produced in the centromere region of whole chromosomes and concentrated and clearly bright signal.The centromeric sequence of the present invention can be directly used for the positioning in function centromere and the identification of correlated series；Genes location of the mark developed available for cotton genetic map centromere position, it is that functional gene positioning and linkage relationship provide more accurate information, also it can offer reference and help for the sequence assembly of Lei Mengdeshi cotton genome sequencings so that its physical map information is more comprehensive.
Description
Technical field
The present invention relates to bioinformatics and molecular cytogenetics field, and in particular to Lei Mengdeshi cotton functions centromere
Sequence and its molecular labeling.
Background technology
Centromere is function element important on higher eucaryote chromosome.In cell mitogen and meiosis
It is responsible for the impartial separation of chromosome in journey, so that playing ensures the important function of inhereditary material stable delivery.Due to centric region
Domain is rich in repetitive sequence, it is difficult to carry out sequencing assembling, therefore, and it is studied falls behind relatively.Although for example, people, drosophila and arabidopsis
The biological genome sequencing of isotype is completed already, but its centromere is because containing a large amount of repetitive sequences, often still with difference
The gap of length is present on gene order-checking collection of illustrative plates, it is impossible to resolved.Therefore, no matter the research in centromere is for its biology
The announcement of characteristic, or the research field such as gene order-checking suffer from significance.
With going deep into that centromere is studied, it has been found that, the centromere with broad sense be the definition of primary constriction on chromosome not
Together, the region generally existing that centromere functions a kind of special histone H 3, i.e. CENH3, therefore, researcher will be true
Centromere in positive meaning is defined as the Chromatin domains combined with the special histone in centromere, i.e. function centric region.With this
Simultaneously as CENH3 defined in centromere in significance, it by as identification centromere functional area and function
The special marking that silk grain DNA sequence dna is excavated.Chromatin immune chemical coprecipitation technique（Chromatin immunoprecipitation
assay, ChIP）It is the main method for studying DNA and protein interaction in organism, therefore, this method also turns into work(
The effective means that energy centromeric sequence is excavated.This method is combined with second generation high throughput sequencing technologies（ChIP-Seq）Can be
Full-length genome level is efficiently separated, analytic function centromeric DNA sequence, has been widely used in function centromere at present
In research.
In recent years, the research and development of cotton gene group is rapid, however, the research in cotton centromere is rarely reported, and only
Report substantially with random DNA sequence analysis, screening based on, it is impossible to determine whether DNA sequence dna is combined with CENH3, therefore
It remains to be discussed as the authenticity of function centromeric sequence.The present invention carries out ChIP-Seq experiments, analysis using CENH3 antibody
Four Lei Mengdeshi cotton function centromeric sequences are obtained, it is clear and definite with CENH3 binding relationships, and passes through entering that FISH is tested
One step is demonstrate,proved, and specify that centromere positioning and its characteristic of function centromeric sequence of these sequences.Meanwhile, for convenience should
With we develop correspondingly special primer according to its sequence, are that its clone and application provide strong instrument.
The content of the invention
The purpose of the present invention is to excavate Lei Mengdeshi cotton function centromeric sequences, and develops its corresponding molecular labeling, is had
Beneficial to cotton genetic map and the structure of physical map, also help centromere and form the discussion that mechanism is exercised with function, also can
Structure enough for cotton artificial chromosome provides required sequence information.
To achieve the above object, the present invention is adopted the following technical scheme that：
Lei Mengdeshi cotton function centromeric sequences, the sequence is obtained according to ChIP-Seq data analyses, and title is respectively：
Sequence 1-Cluster201Contig81, sequence 2-Cluster285Contig8, sequence 3-Cluster291Contig8, sequence
4-Cluster407Contig2, its nucleotide sequence is respectively shown in SEQ ID NO.1-4.
Described sequence 1-Cluster201Contig81, sequence 2-Cluster285Contig8, sequence 3-
The corresponding molecular labeling primer sequences of Cluster291Contig8 and sequence 4-Cluster407Contig2 such as SEQ ID
Shown in NO.5-12.
The corresponding molecular labeling of Lei Mengdeshi cotton function centromeric sequences of table 1
1st, Lei Mengdeshi cottons function centromeric sequence
The Lei Mengdeshi cotton function centromeric sequences of the present invention are obtained using ChIP-Seq data analyses.First with
CENH3-DNA complexs are enriched with to designed, designed and available CENH3 antibody specificities, then purifying DNA fragment, and through end
Repair, add A and add after sequence measuring joints, then the amplifications of the PCR through low-circulation number and agarose gel electrophoresis reclaim particular size fragment,
Complete work prepared by sequencing library.It is sequenced using Illumina Hiseq-2500 sequenators, and passes through bioinformatics
Analysis, identifies the site that CENH3 is combined with genomic DNA, and the DNA sequence dna clearly combined closely with CENH3.
2nd, the design of primer and its PCR amplifications
Lei Mengdeshi cottons function centromeric sequence of the present invention has following points for attention when designing primer：One is that primer should
With the conservative interior design of nucleotide sequence and with specificity；Two be that primer length is generally 16-25 bp；Three be the GC of primer sequence
Content is generally 40-60%；Four be the base random distribution of primer sequence；Five be that PCR primer size is 200-700 bp.
PCR amplification condition be：95 DEG C of pre-degeneration 3min, 95 DEG C of 30s, 47-53 DEG C of 30s, 72 DEG C of 45s, 35 are followed
Ring, 72 DEG C of overall elongation 7min.It should be noted that primer sequence is different, annealing temperature is otherwise varied：Sequence 1 is 50 DEG C, sequence
2 be 47 DEG C, sequence 3 is 53 DEG C, sequence 4 is 51 DEG C.The band of PCR amplifications is single and bright, therefore directly uses QIAGEN reagents
Box carries out PCR primer purifying and reclaimed, its article No.：28104, title：QIAquick PCR Purification Kit(50).
The advantage of the invention is that：
Definition of the invention according to function centromere, the special histone CENH3 in the available cotton centromere of designed, designed is opened
Open up ChIP experiments, and DNA sequence dna progress high-flux sequence by enrichment and after purification.This is to obtain having for function centromeric sequence
Efficacious prescriptions method.By bioinformatic analysis, the contigs combined with CENH3 with high abundance has been obtained, and it is real by FISH
Test, demonstrate the centromere positioning of sequence, show that the Lei Mengdeshi cotton functions centromeric sequence obtained by the present invention is true and reliable,
It efficiently avoid and the uncertainty that centromeric sequence is sought is carried out only from DNA sequence dna level.We are according to different sequences
Corresponding PCR primer is designed, and primer specificity is good, and the band of amplification is single and bright, therefore is suitable for purifying PCR productions
Thing, label probe, for FISH experiments.The clone for being designed as function centromeric sequence of PCR primer and application provide important
Instrument.The present invention have accumulated valuable materials for the research of cotton function centromeric sequence, be conducive to cotton genetic map and physics
The structure of collection of illustrative plates with it is perfect, also help centromere formed with function exercise mechanism discussion, also can for cotton it is artificially colored
The structure of body provides required sequence information.
Brief description of the drawings
Fig. 1：Results of hybridization-the A of Lei Mengdeshi cotton function centromeric sequences：Lei Mengdeshi cotton mitosis metaphases are dyed
Body, B：Centromere regional signal, C：Composite diagram.From the figure, it can be seen that the signal that sequence of the present invention is produced is in Lei Mengdeshi
The centromere region of cotton colour solid, demonstrates the centromere positioning of sequence.
Embodiment
Embodiment 1：The implementation of ChIP-Seq technologies
ChIP-Seq technologies refer to chromatin immune co-precipitation ChIP and are combined with new-generation sequencing, and this method is feasible, stably,
The research of individual gene and protein binding relation is applicable not only to, the full-length genome for applying also for differential protein binding site is ground
Study carefully.
ChIP-Seq basic procedure is used in of the invention：
1st, liquid nitrogen grinding is fresh and tender blade；2nd, nucleus, micrococcus luteus ribozyme enzymolysis are extracted；3rd, centromere is added special
Different histone CENH3 antibody, be combined with each other with target protein-DNA compounds；4th, ProteinA, binding antibody-target egg are added
- DNA compounds, and precipitating in vain；5th, the compound precipitated is cleaned, removes some non-specific bindings；6th, elute,
Target protein-DNA the compounds being enriched with；7th, solution crosslinking, the DNA segment of purification enrichment；8th, PCR is analyzed and is prepared sequencing text
Storehouse；9th, it is sequenced using Illumina Hiseq-2500 sequenators；10th, by initial data（reads）Carry out bioinformatics
Analysis, identifies the site that CENH3 is combined with genomic DNA, and the DNA sequence dna clearly combined closely with CENH3.
ChIP-Seq concrete operations flow is：
（1）Liquid nitrogen grinding is fresh and tender blade, extracts nucleus；
（2）Using micrococcus luteus ribozyme lysed cells core, chromatin is interrupted at random for 200-1000 bp small fragment；
（3）Nucleus after enzymolysis is divided into three parts, portion is internal reference（input）, portion addition preimmune serum
（Mock）, another adds the specific antibody mediated immunity precipitating proteins-DNA complexs 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 standing 10min, 700rpm centrifugation 1min, remove supernatant.
（6）Successively sediment composite is cleaned with low salt solutions, high level salt solution, LiCl and TE.Cleaning step is：Add molten
Liquid, 4 DEG C of tops turn 10min, 4 DEG C of standing 10min, 700rpm centrifugation 1min, remove supernatant.
（7）After cleaning is finished, start elution.Elute formula of liquid：1ml solution compositions are：100 μ L10%SDS, 100 μ L1M
NaHCO3, 800 μ LddH2O.Often pipe adds eluent, and room temperature top turns 15min, stood after centrifugation, collects supernatant.Repeated washing one
It is secondary.
（8）Solution crosslinking：Often pipe adds NaCl, makes its final concentration of 0.2M.Mix, 65 DEG C of solution crosslinkings are stayed overnight.
（9）After solution crosslinking terminates, often pipe adds RNaseA, 37 DEG C of incubation 1h.
（10）Often pipe adds EDTA, Tris-HCl and Proteinase K, 45 DEG C of processing 2h.
（11）The recovery of DNA fragmentation：Using QIAGEN glue reclaim kits, article No.：28704, title：QIAquick Gel
Extraction Kit (50)。
（12）Obtained DNA is purified to repair through end plus A and after adding sequence measuring joints, then the amplifications of the PCR through low-circulation number and
Agarose gel electrophoresis reclaims particular size fragment, prepares sequencing library.
（13）It is sequenced using Illumina Hiseq-2500 sequenators.
（14）Using the method for bioinformatics, the reads that sequencing is obtained is spliced after carrying out quality control, is obtained
Different contigs；By being compared with reference gene group, ambient interferences are removed, being tied with CENH3 with high abundance is obtained
The contigs of conjunction, i.e. function centromeric sequence.In order to further confirm that the centromere of the sequence is positioned, we are miscellaneous using FISH
The method of friendship, verifies the centromere positioning of these sequences, so that clear and definite its is function centromeric sequence.
According to the definition in function centromere, if with CENH3 protein bindings be to judge DNA sequence dna whether as function centromere
The core condition of sequence.Therefore, will come from function centromere using the CENH3 sequences obtained by the ChIP experiments of means
DNA sequence dna.Because metaphase chromosome FISH hybridizes the limitation of resolution ratio, the existing so-called centromeric DNA sequence of cotton is first
First can not be clear and definite whether in centric region, secondly clear and definite whether it can not also be bound with CENH3, therefore can not determine whether it is
Function centromeric DNA sequence, the present invention is using ChIP-Seq method, and therefore, its sequence comes from cotton function centric region,
FISH experiments also demonstrate the centromere positioning of the validity and sequence of this method.
Embodiment 2：Design of primers is expanded with PCR
Design of primers is carried out to function centromeric sequence using primer-design software Primer Premier 5.0, bar is set
Part is：One is the total length or partial sequence of nucleotide sequence；Two be that primer length is the bp of 21bp ± 5；Three be the GC of primer sequence
Content is 40-60%；Four be that PCR primer size is 200-700 bp.
PCR amplification condition be：95 DEG C of pre-degeneration 3min, 95 DEG C of 30s, 47-53 DEG C of 30s, 72 DEG C of 45s, 35 are followed
Ring, 72 DEG C of overall elongation 7min.It should be noted that primer sequence is different, annealing temperature is otherwise varied：Sequence 1 is 50 DEG C, sequence
2 be 47 DEG C, sequence 3 is 53 DEG C, sequence 4 is 51 DEG C.The band of PCR amplifications is single and bright.Therefore directly tried using QIAGEN
Agent box carries out PCR primer purifying and reclaimed, its article No.：28104, title：QIAquick PCR Purification Kit(50).
The foregoing is only presently preferred embodiments of the present invention, all equivalent changes done according to scope of the present invention patent with
Modification, should all belong to the covering scope of the present invention.
SEQUENCE LISTING
<110>University Of Agriculture and Forestry In Fujian
<120>Lei Mengdeshi cotton function centromeric sequences and its molecular labeling
<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 (3)
1. the molecular labeling of Lei Mengdeshi cotton function centromeric sequences, it is characterised in that：The Lei Mengdeshi cottons function centromere
Sequence 1-Cluster201Contig81, sequence 2-Cluster285Contig8, sequence 3-Cluster291Contig8 and sequence
Arrange 4-Cluster407Contig2 corresponding molecular labeling primer sequences respectively as SEQ ID NO.5-6, SEQ ID NO.7-8,
Shown in SEQ ID NO.9-10, SEQ ID NO.11-12；
Sequence 1-Cluster201Contig81, sequence 2-Cluster285Contig8, sequence 3-Cluster291Contig8,
Sequence 4-Cluster407Contig2, its nucleotide sequence is respectively shown in SEQ ID NO.1-4.
2. a kind of molecular labeling determining in function centromere of Lei Mengdeshi cottons function centromeric sequence as claimed in claim 1
Application in the identification of position and Lei Mengdeshi cotton function centromeric sequences.
3. a kind of molecular labeling of Lei Mengdeshi cottons function centromeric sequence as claimed in claim 1 in cotton genetic map
Application on the Genes location of silk grain position.
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| Song Luo.The Cotton Centromere Contains a Ty3-gypsy-like LTR Retroelement.《PloS One》.2012,第7卷(第4期),e35261. * |
| Zhiyun Gong.Repeatless and Repeat-Based Centromeres in Potato: Implications for Centromere Evolution.《The Plant Cell》.2012,第24卷3559–3574. * |
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