CN110183523A - OsMYB36 albumen and its encoding gene and application - Google Patents
OsMYB36 albumen and its encoding gene and application Download PDFInfo
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- CN110183523A CN110183523A CN201910462534.9A CN201910462534A CN110183523A CN 110183523 A CN110183523 A CN 110183523A CN 201910462534 A CN201910462534 A CN 201910462534A CN 110183523 A CN110183523 A CN 110183523A
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- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
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- C12N15/8261—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield
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Abstract
The invention discloses OsMYB36 albumen and its encoding gene and applications.Protein provided by the invention is named as OsMYB36 albumen, is obtained from rice (Oryza sativa), is protein shown in sequence 1 in sequence table.The nucleic acid molecules of coding OsMYB36 albumen also belong to protection scope of the present invention.A kind of method that the present invention also protects prepare transgenosis plant includes the following steps: to import the substance for inhibiting the nucleic acid molecules expression in recipient plant, obtains genetically modified plants；Compared with the recipient plant, the transgenic plant has at least one of following four kinds of phenotypes: plant height increases, and fringe increases, and seed grain length increases, the wide reduction of seed grain.The present invention can be used for a variety of character improvements of plant, have great application and popularization value for plant breeding.
Description
Technical field
The present invention relates to OsMYB36 albumen and its encoding gene and applications.
Background technique
Rice is native to China and India.It is one of world's staple food crop.Rice in China sowing face accounts for national grain
The 1/4 of crop, and yield then accounts for more than half.Cultivation history is 14000~18000 years existing.For important cereal crops；Except edible
Outside caryopsis, can starch processed, make wine, vinegar processed, rice bran can refine sugar, extracts oil, extract furfural, for industry and pharmaceutical；Rice straw is good
Feed and paper making raw material and braided material, rice sprout and rice root can hyoscines.
In recent years, the rapid development of biotechnology has greatly pushed the innovation and research level of plant breeding research means
Continuous improvement, Genes For Plant Tolerance disease pest, antiweed biotechnology breeding have initially entered practical stage.Utilize biotechnology hand
Section imports external source desinsection, anti-herbicide gene in Plant Genome, has broken and has been difficult to hybridize between plant species even species
Natural barrier, realize pest-resistant, anti-herbicide gene transfer, thus make plant promptly, directionally obtain insect resistace and
Mechanization weeding, while original good economical character can be retained again.Since every plant of plant of transgenic corns all has quite
The resistance of degree, thus its is pest-resistant, antiweed effect well and is stablized than artificial control's control efficiency, additionally it is possible to save manpower
With the investment of material resources, social resources are effectively saved.Other economical characters improve genetically modified plants progress and application not as good as it is pest-resistant,
Herbicide resistance is ideal, and mainly due to unexcellent character improvement gene, most economical character mainly has one slightly
Caused by the controlled by multiple genes of effect, never ideal gene is operated.
Summary of the invention
The object of the present invention is to provide OsMYB36 albumen and its encoding gene and applications.
Protein provided by the invention is named as OsMYB36 albumen, is obtained from rice (Oryza sativa), is as follows
(a1) or (a2) or (a3) or (a4):
(a1) protein shown in sequence 1 in sequence table；
(a2) fusion protein obtained in N-terminal or/and C-terminal the connection label of (a1) described protein；
(a3) by (a1) by one or several amino acid residues substitution and/or deletion and/or addition obtain with plant
Object develops relevant protein；
(a4) there is 98% or more identity and protein relevant with development of plants from rice and to (a1).
The development of plants is presented as plant height and/or fringe character and/or grain characters.
Label is specifically as shown in table 1.
The sequence of 1 label of table
The nucleic acid molecules of coding OsMYB36 albumen also belong to protection scope of the present invention.
The nucleic acid molecules are concretely following (b1) or (b2) or (b3) or (b4):
(b1) code area DNA molecular as shown in sequence 2 in sequence table；
(b2) DNA molecular shown in sequence 3 in sequence table；
(b3) there are the DNA of 95% or more identity and code for said proteins points from rice and with (b1) or (b2)
Son；
(b4) under strict conditions with (b1) or (b2) limit nucleotide sequence hybridization and code for said proteins DNA
Molecule.
Above-mentioned stringent condition can for 6 × SSC, the solution of 0.5%SDS hybridizes at 65 DEG C, then with 2 × SSC,
It is primary that 0.1%SDS and 1 × SSC, 0.1%SDS respectively wash film.
Expression cassette, recombinant vector or recombinant microorganism containing the nucleic acid molecules all belong to the scope of protection of the present invention.
The present invention also protects the application of OsMYB36 albumen, for as follows (c1), (c2), (c3) or (c4):
(c1) regulate and control the plant height character of plant；
(c2) regulate and control the fringe character of plant；
(c3) regulate and control the seed grain length character of plant；
(c4) regulate and control the wide character of seed grain of plant.
(c1) described to be regulated to negative regulation in, even if plant plant height reduces.
(c2) described to be regulated to negative regulation in, even if plant fringe reduces.
(c3) described to be regulated to negative regulation in, even if plant seed grain length reduces.
(c4) in, the regulation is positive regulation, even if the wide increase of plant seed grain.
The present invention also protects the application of the nucleic acid molecules, for as follows (d1), (d2), (d3) or (d4):
(d1) genetically modified plants that plant height character changes are cultivated；
(d2) genetically modified plants that fringe character changes are cultivated；
(d3) genetically modified plants that seed grain length character changes are cultivated；
(d4) genetically modified plants that the wide character of seed grain changes are cultivated.
(d1) in, the plant height character changes into plant height reduction.
(d2) in, the fringe character changes into fringe reduction.
(d3) in, the seed grain length character changes into the reduction of seed grain length.
(d4) in, the wide character of seed grain changes into the wide increase of seed grain.
The present invention also protects the substance for inhibiting OsMYB36 albumen in the application of plant breeding, and the target of the breeding is such as
Under (e1), (e2), (e3) or (e4):
(e1) plant that plant height increases is cultivated；
(e2) plant that fringe increases is cultivated；
(e3) the increased plant of seed grain length is cultivated；
(e4) the wide reduced plant of seed grain is cultivated.
The present invention also protects the application for inhibiting the substance of the nucleic acid molecules, for as follows (f1), (f2), (f3) or (f4):
(f1) genetically modified plants that plant height increases are cultivated；
(f2) genetically modified plants that fringe increases are cultivated；
(f3) the increased genetically modified plants of seed grain length are cultivated；
(f4) the wide reduced genetically modified plants of seed grain are cultivated.
A kind of method that the present invention also protects prepare transgenosis plant includes the following steps: to import suppression in recipient plant
The substance for making the nucleic acid molecules expression, obtains genetically modified plants；Compared with the recipient plant, the transgenic plant has
At least one of following four kinds of phenotypes: plant height increases, and fringe increases, and seed grain length increases, the wide reduction of seed grain.With the receptor
Plant is compared, and the transgenic plant has any one in following four kinds of phenotypes: plant height increases, and fringe increases, seed grain length
Increase, the wide reduction of seed grain.Compared with the recipient plant, the transgenic plant has following four kinds of phenotypes: plant height simultaneously
Increase, fringe increases, and seed grain length increases, the wide reduction of seed grain.
The substance of the nucleic acid molecules is inhibited concretely to pass through the substance that gene editing inhibits the nucleic acid molecules.Inhibit
The substance of the nucleic acid molecules concretely Cas9 system.In Cas9 system, target sequence binding region such as sequence table in sgRNA
Shown in sequence 6.In Cas9 system, the coded sequence of sgRNA is as shown in the sequence 5 of sequence table.Inhibit the object of the nucleic acid molecules
Matter concretely has the recombinant plasmid of the coded sequence of Cas9 albumen and the coded sequence of special sgRNA.In special sgRNA
Target sequence binding region is as shown in the sequence 6 of sequence table.Special sgRNA is specially the corresponding RNA of DNA shown in the sequence 5 of sequence table.
In the specific sequence 4 such as sequence table of the coded sequence of Cas9 albumen shown in 2697-6968 nucleotide.Inhibit the nucleic acid point
The substance concretely recombinant plasmid SG2027 of son.
The present invention also protects a kind of plant breeding method, includes the following steps: to reduce OsMYB36 albumen in purpose plant
Content and/or activity, to make purpose plant that at least one of following four kinds of phenotypic alternations occur: plant height increases, and fringe increases,
Seed grain length increases, the wide reduction of seed grain.To make purpose plant that any one in following four kinds of phenotypic alternations: plant height occur
Increase, fringe increases, and seed grain length increases, the wide reduction of seed grain.To make purpose plant while following four kinds of phenotypic alternations occur:
Plant height increases, and fringe increases, and seed grain length increases, the wide reduction of seed grain.
Any description above plant height is strain plant height.
Any description above plant is monocotyledon or dicotyledon.Any description above plant is grass family plant
Object.Any description above plant is oryza plant.Any description above plant is rice, such as rice OryzasativaLcv.Nipponbare.
The present invention can be used for a variety of character improvements of plant, have great application and popularization value for plant breeding.
Detailed description of the invention
Fig. 1 is the structural schematic diagram of recombinant plasmid SG5752.
Fig. 2 is the sequencing result of mutational site and its periphery nucleotide.
Fig. 3 is the photo of maturity period plant.
Fig. 4 is the plant height of maturity period plant.
Fig. 5 is the photo of mature spike of rice.
Fig. 6 is the spike length of mature spike of rice.
Fig. 7 is Primary branch quantity and Secondary branch quantity.
Fig. 8 is the photo of ripe seed.
Fig. 9 is the result of Ear weight.
Figure 10 is grain length and the wide result of grain.
Specific embodiment
Embodiment below facilitates a better understanding of the present invention, but does not limit the present invention.Experiment in following embodiments
Method is unless otherwise specified conventional method.Test material as used in the following examples is unless otherwise specified certainly
What routine biochemistry reagent shop was commercially available.Quantitative test in following embodiment is respectively provided with three repeated experiments, as a result makes even
Mean value.
Embodiment,
OsMYB36 albumen is as shown in the sequence 1 of sequence table.In the cDNA of rice OryzasativaLcv.Nipponbare, coding OsMYB36 albumen
CDS is as shown in the sequence 2 of sequence table.In the genomic DNA of rice OryzasativaLcv.Nipponbare, the gene such as sequence table of OsMYB36 albumen is encoded
Sequence 3 shown in.
One, construction recombination plasmid
Construction recombination plasmid SG5752.The structural schematic diagram of recombinant plasmid SG5752 is shown in Fig. 1.Through full plasmid order-checking, recombination
Plasmid SG5752 is as shown in the sequence 4 of sequence table.In the sequence 4 of sequence table, 2697-6968 nucleotide coding Cas9 eggs
It is white.In recombinant plasmid SG5752, the code area of sgRNA (96bp) as shown in the sequence 5 of sequence table.Target sequence knot in sgRNA
Area is closed as shown in the sequence 6 of sequence table.
Two, it carries out genetic transformation and obtains regeneration plant
Recombinant plasmid SG5752 is imported into Agrobacterium EHA105, obtains recombinational agrobacterium.Using During Agrobacterium method, will weigh
Group Agrobacterium carries out genetic transformation to the embryo callus of rice OryzasativaLcv.Nipponbare, then screens resistant calli (resistance screening
Using 100mg/L hygromycin), differentiation and regeneration culture is then carried out, culture of rootage is then carried out, obtains regeneration plant.
Three, transgenic plant and its Progeny plants are obtained
The regeneration plant that step 2 obtains is identified as follows: taking blade, genomic DNA is extracted, using primer
The primer pair of GP5752-4619-F and primer GP5752-4619-R composition carries out PCR amplification, then carries out pcr amplification product
Sequencing.
GP5752-4619-F:5 '-AGCACCATGACCAAACCCAT-3 '；
GP5752-4619-R:5 '-CCTCTTCAGCCCTGCAACAT-3 '.
By identifying above, screening obtains two homozygous mutant plant (i.e. two from the regeneration plant that step 2 obtains
The mutation that chromosome occurs is consistent), it is respectively designated as osmyb36-1 plant and osmyb36-2 plant.
It is identified through sequencing, compared with the genomic DNA of rice OryzasativaLcv.Nipponbare (being indicated with Nip), the difference of osmyb36-1 plant
It is only that the insertion that a nucleotide has occurred in the gene of coding OsMYB36 albumen, the mutational site and its periphery nucleosides
The sequencing result of acid is shown in Fig. 2.
It is identified through sequencing, compared with the genomic DNA of rice OryzasativaLcv.Nipponbare (being indicated with Nip), the difference of osmyb36-2 plant
It is only that missing (target sequence of the missing in sgRNA that a nucleotide has occurred in the gene of coding OsMYB36 albumen
In).
Osmyb36-1 plant selfing simultaneously harvests seed, is plant by cultivating seeds, as T1 is for plant.T1 for plant from
Seed is handed over and harvested, is plant by cultivating seeds, as T2 is for plant.Osmyb36-1 plant and its self progeny, referred to as
Osmyb36-1 strain.
Osmyb36-2 plant selfing simultaneously harvests seed, is plant by cultivating seeds, as T1 is for plant.T1 for plant from
Seed is handed over and harvested, is plant by cultivating seeds, as T2 is for plant.Osmyb36-2 plant and its self progeny, referred to as
Osmyb36-2 strain.
Four, character compares
For try plant are as follows: rice OryzasativaLcv.Nipponbare (being indicated with Nip), osmyb36-1 strain T2 for plant, osmyb36-2 plants
The T2 of system is for plant.
Culture is for trying plant under parallel condition.
The photo of maturity period plant is shown in Fig. 3.The plant height of maturity period plant is shown in that (each strain takes being averaged for 10 plants of plant to Fig. 4
Value).Compared with rice OryzasativaLcv.Nipponbare, osmyb36-1 strain plant and the equal plant height of osmyb36-2 strain plant significantly increase.
The plant height of osmyb36-1 strain plant and osmyb36-2 strain plant is without significant difference.
The photo of mature spike of rice is shown in Fig. 5.The spike length of mature spike of rice is shown in Fig. 6 (average value that each strain takes 10 plants of plant).
Primary branch quantity and Secondary branch quantity are shown in Fig. 7.Compared with rice OryzasativaLcv.Nipponbare, osmyb36-1 strain plant and osmyb36-2
The equal spike length of strain plant dramatically increases.The spike length of osmyb36-1 strain plant and osmyb36-2 strain plant is without significant difference.
The photo of ripe seed is shown in Fig. 8 (upper figure is the seed that do not shell, and the following figure is shelling seed).The result of Ear weight is shown in figure
9 (average values that each strain takes 10 plants of plant).(each strain takes being averaged for 10 plants of plant for grain length and the wide the result is shown in Figure 10 of grain
Value).Compared with rice OryzasativaLcv.Nipponbare, osmyb36-1 strain plant and the equal grain length of osmyb36-2 strain plant are dramatically increased.
The grain length of osmyb36-1 strain plant and osmyb36-2 strain plant is without significant difference.Compared with rice OryzasativaLcv.Nipponbare,
The osmyb36-1 strain plant and equal grain of osmyb36-2 strain plant is wide is substantially reduced.Osmyb36-1 strain plant and
The wide no significant difference of the grain of osmyb36-2 strain plant.
The data result of above each character is shown in Table 2.
Table 2
| Rice OryzasativaLcv.Nipponbare | Osmyb36-1 strain plant | Osmyb36-2 strain plant | |
| Plant height | 72.6cm | 91.3cm | 91.1cm |
| Spike length | 17.5cm | 22.8cm | 23.1cm |
| Primary branch quantity | 6.9 | 7.3 | 7.2 |
| Secondary branch quantity | 11.9 | 18.3 | 18.5 |
| Ear weight | 1.22g | 2.24g | 2.23g |
| Grain length | 0.68cm | 0.81cm | 0.82cm |
| Grain is wide | 0.32cm | 0.26cm | 0.25cm |
SEQUENCE LISTING
<110>Biological Technology institute, Chinese Academy of Agricultural Sciences
<120>OsMYB36 albumen and its encoding gene and application
<130> GNCYX191205
<160> 6
<170> PatentIn version 3.5
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ctgcctcaca agattgggct gaagaggtgt ggcaagagct gcaggctaag gtggctgaat 180
tacctgaggc caaacataaa gcatggggac ttcaccccag aggaggacag catcatctgc 240
agcctctaca ttagcatagg gagcaggtgg tcaatcatag cagcacagct gccagggagg 300
acggacaacg atgtcaagaa ctactggaac acaaagctga agaagagact ccttggccgg 360
cgcaaggacc gcggcggcgg ccaccaccac cgcagccaga gcaccgccga cgatcttccg 420
gccggtggtg acggcggcat gaacgacggc ggcggcggcg gcggagagcg gtcgctgagc 480
gcgtcggcga tggagaggat ccagctctgc atgcagctgc aggagctgca gaacccactg 540
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tcgccgtcga gcggcggcga gaactccaac gtcgtcgtcg ccatcgaggc cgagctccag 780
gagcttctct acgccggcgg cggcgcgatc gtcgacggcg gcgcgccgcc gcagggggat 840
gtggactggt ggagctatga tcagggaaag cagtcacctg tgacttgctg ggatttcacc 900
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aaacagcatg gtttggtatt ccatctagct atgaacattt tttttttcta tcatggccca 180
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attccatact attattacta ctacattatt ataattaaaa aattaataga atatcaagat 780
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gagttcagag tacttactgt tcttcgtgat ctgtgcactc ctaatctctc ctaaacccac 1140
atgtagctct aaatatatag tgccattttg aatgcatttt ctcgcatcca ccgtttagat 1200
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aacaatgatt attttgcaga gaacagttag gagacagaaa tccatgtccc agagaacaaa 1620
aaagaggaca tatatatgca tggatgacac agaagtttag caaaaagcac aacaaatcat 1680
tccatgcacc tttctgcatg cattcatgca atgcagaaac aaaaaggcca tgcatgcatc 1740
tccaaagtac caatagacca gaacaagagc cggtatgtgt gtcatgctct ttcagcagca 1800
tcagcaatca aaggtcaaac catatccgta ctattttaat gcactcttca tctgctccta 1860
taaatcccag caccatgacc aaacccatca caactcatct catcacattc catacacaca 1920
tccatagcag cttcaagcta aggtgatact actacattca gagagaagaa gcagaaagag 1980
ataagaggtt ctgagagaga ctgagagaga tggggagggc accttgctgt gacaaggcaa 2040
cagtgaagaa ggggccatgg tcacctgagg aggatgcaat gctcaagaac tacattgagg 2100
agcatggcac cggtggcaac tggattgcac tgcctcacaa gattggtagc aattattacc 2160
atttttactt atcactgata gtttcagaag agattataaa ttttcagaca tgctaagatt 2220
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gtgtaatgtt gcagggctga agaggtgtgg caagagctgc aggctaaggt ggctgaatta 2400
cctgaggcca aacataaagc atggggactt caccccagag gaggacagca tcatctgcag 2460
cctctacatt agcataggga gcaggtacat aattacacca ctaaccactc tcccttctca 2520
ctaactatca atcaatcatc ataatcacac taaacttgac ataattaacc acaggtggtc 2580
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aaagctgaag aagagactcc ttggccggcg caaggaccgc ggcggcggcc accaccaccg 2700
cagccagagc accgccgacg atcttccggc cggtggtgac ggcggcatga acgacggcgg 2760
cggcggcggc ggagagcggt cgctgagcgc gtcggcgatg gagaggatcc agctctgcat 2820
gcagctgcag gagctgcaga acccactgtc catccaccac aaccccttgc tctctcatca 2880
gtggccaagc aaggccacca ttgatgatca gaatcacaac aatgtcactg tggctgaaca 2940
tggaatgtca agctctgtga gcgaccacca ccgcctcgat gggcagcagc tggagagcgg 3000
cgccggcgcc gccgccatgc agcaggcgtc gccgtcgagc ggcggcgaga actccaacgt 3060
cgtcgtcgcc atcgaggccg agctccagga gcttctctac gccggcggcg gcgcgatcgt 3120
cgacggcggc gcgccgccgc agggggatgt ggactggtgg agctatgatc agggaaagca 3180
gtcacctgtg acttgctggg atttcacccc tgaaaccagc tccatcttcc aggattatgc 3240
aacagtttat gacatctgaa aattgtgaag aggtttgata aggatgttaa ttagggaggt 3300
aataataata ataatataaa tagggattgt ctagcctttg tgttctttga tcagtgacac 3360
ctaaaaccaa actgtaagga gtttgttgta agaaatttgt agcaagggat aaataaaaag 3420
gtatggtaaa aaatatatat accaggtcca agagatgatg taatcttggc accattctgt 3480
caaagatcag ataggactga tgagctcaga taaaaatgca aggtggcttt cactggaaga 3540
agaaaaagga ctttatatcc tcacaatagt cactttcatg catggcctct gcgacaatat 3600
tccctggtgt tacaatca 3618
<210> 4
<211> 15898
<212> DNA
<213> Artificial sequence
<400> 4
taaacgctct tttctcttag gtttacccgc caatatatcc tgtcaaacac tgatagttta 60
aactgaaggc gggaaacgac aatctgatcc aagctcaagc tgctctagca ttcgccattc 120
aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg 180
gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca 240
cgacgttgta aaacgacggc cagtgccaag cttggatcat gaaccaacgg cctggctgta 300
tttggtggtt gtgtagggag atggggagaa gaaaagcccg attctcttcg ctgtgatggg 360
ctggatgcat gcgggggagc gggaggccca agtacgtgca cggtgagcgg cccacagggc 420
gagtgtgagc gcgagaggcg ggaggaacag tttagtacca cattgcccag ctaactcgaa 480
cgcgaccaac ttataaaccc gcgcgctgtc gcttgtgtta cattgaggag catggcacgt 540
tttagagcta gaaatagcaa gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg 600
caccgagtcg gtgctttttt gttttagagc tagaaatagc aagttaaaat aaggctagtc 660
cgtagcgcgt gcgccaattc tgcagacaaa tggccccggg cctgcaggtg cagcgtgacc 720
cggtcgtgcc cctctctaga gataatgagc attgcatgtc taagttataa aaaattacca 780
catatttttt ttgtcacact tgtttgaagt gcagtttatc tatctttata catatattta 840
aactttactc tacgaataat ataatctata gtactacaat aatatcagtg ttttagagaa 900
tcatataaat gaacagttag acatggtcta aaggacaatt gagtattttg acaacaggac 960
tctacagttt tatcttttta gtgtgcatgt gttctccttt ttttttgcaa atagcttcac 1020
ctatataata cttcatccat tttattagta catccattta gggtttaggg ttaatggttt 1080
ttatagacta atttttttag tacatctatt ttattctatt ttagcctcta aattaagaaa 1140
actaaaactc tattttagtt tttttattta ataatttaga tataaaatag aataaaataa 1200
agtgactaaa aattaaacaa atacccttta agaaattaaa aaaactaagg aaacattttt 1260
cttgtttcga gtagataatg ccagcctgtt aaacgccgtc gacgagtcta acggacacca 1320
accagcgaac cagcagcgtc gcgtcgggcc aagcgaagca gacggcacgg catctctgtc 1380
gctgcctctg gacccctctc gagagttccg ctccaccgtt ggacttgctc cgctgtcggc 1440
atccagaaat tgcgtggcgg agcggcagac gtgagccggc acggcaggcg gcctcctcct 1500
cctctcacgg cacggcagct acgggggatt cctttcccac cgctccttcg ctttcccttc 1560
ctcgcccgcc gtaataaata gacaccccct ccacaccctc tttccccaac ctcgtgttgt 1620
tcggagcgca cacacacaca accagatctc ccccaaatcc acccgtcggc acctccgctt 1680
caaggtacgc cgctcgtcct cccccccccc ccctctctac cttctctaga tcggcgttcc 1740
ggtccatggt tagggcccgg tagttctact tctgttcatg tttgtgttag atccgtgttt 1800
gtgttagatc cgtgctgcta gcgttcgtac acggatgcga cctgtacgtc agacacgttc 1860
tgattgctaa cttgccagtg tttctctttg gggaatcctg ggatggctct agccgttccg 1920
cagacgggat cgatttcatg attttttttg tttcgttgca tagggtttgg tttgcccttt 1980
tcctttattt caatatatgc cgtgcacttg tttgtcgggt catcttttca tgcttttttt 2040
tgtcttggtt gtgatgatgt ggtctggttg ggcggtcgtt ctagatcgga gtagaattct 2100
gtttcaaact acctggtgga tttattaatt ttggatctgt atgtgtgtgc catacatatt 2160
catagttacg aattgaagat gatggatgga aatatcgatc taggataggt atacatgttg 2220
atgcgggttt tactgatgca tatacagaga tgctttttgt tcgcttggtt gtgatgatgt 2280
ggtgtggttg ggcggtcgtt cattcgttct agatcggagt agaatactgt ttcaaactac 2340
ctggtgtatt tattaatttt ggaactgtat gtgtgtgtca tacatcttca tagttacgag 2400
tttaagatgg atggaaatat cgatctagga taggtataca tgttgatgtg ggttttactg 2460
atgcatatac atgatggcat atgcagcatc tattcatatg ctctaacctt gagtacctat 2520
ctattataat aaacaagtat gttttataat tattttgatc ttgatatact tggatgatgg 2580
catatgcagc agctatatgt ggattttttt agccctgcct tcatacgcta tttatttgct 2640
tggtactgtt tcttttgtcg atgctcaccc tgttgtttgg tgttacttct gcagccatgg 2700
actataagga ccacgacgga gactacaagg atcatgatat tgattacaaa gacgatgacg 2760
ataagatggc cccaaagaag aagcggaagg tcggtatcca cggagtccca gcagccgaca 2820
agaagtacag catcggcctg gacatcggca ccaactctgt gggctgggcc gtgatcaccg 2880
acgagtacaa ggtgcccagc aagaaattca aggtgctggg caacaccgac cggcacagca 2940
tcaagaagaa cctgatcgga gccctgctgt tcgacagcgg cgaaacagcc gaggccaccc 3000
ggctgaagag aaccgccaga agaagataca ccagacggaa gaaccggatc tgctatctgc 3060
aagagatctt cagcaacgag atggccaagg tggacgacag cttcttccac agactggaag 3120
agtccttcct ggtggaagag gataagaagc acgagcggca ccccatcttc ggcaacatcg 3180
tggacgaggt ggcctaccac gagaagtacc ccaccatcta ccacctgaga aagaaactgg 3240
tggacagcac cgacaaggcc gacctgcggc tgatctatct ggccctggcc cacatgatca 3300
agttccgggg ccacttcctg atcgagggcg acctgaaccc cgacaacagc gacgtggaca 3360
agctgttcat ccagctggtg cagacctaca accagctgtt cgaggaaaac cccatcaacg 3420
ccagcggcgt ggacgccaag gccatcctgt ctgccagact gagcaagagc agacggctgg 3480
aaaatctgat cgcccagctg cccggcgaga agaagaatgg cctgttcgga aacctgattg 3540
ccctgagcct gggcctgacc cccaacttca agagcaactt cgacctggcc gaggatgcca 3600
aactgcagct gagcaaggac acctacgacg acgacctgga caacctgctg gcccagatcg 3660
gcgaccagta cgccgacctg tttctggccg ccaagaacct gtccgacgcc atcctgctga 3720
gcgacatcct gagagtgaac accgagatca ccaaggcccc cctgagcgcc tctatgatca 3780
agagatacga cgagcaccac caggacctga ccctgctgaa agctctcgtg cggcagcagc 3840
tgcctgagaa gtacaaagag attttcttcg accagagcaa gaacggctac gccggctaca 3900
ttgacggcgg agccagccag gaagagttct acaagttcat caagcccatc ctggaaaaga 3960
tggacggcac cgaggaactg ctcgtgaagc tgaacagaga ggacctgctg cggaagcagc 4020
ggaccttcga caacggcagc atcccccacc agatccacct gggagagctg cacgccattc 4080
tgcggcggca ggaagatttt tacccattcc tgaaggacaa ccgggaaaag atcgagaaga 4140
tcctgacctt ccgcatcccc tactacgtgg gccctctggc caggggaaac agcagattcg 4200
cctggatgac cagaaagagc gaggaaacca tcaccccctg gaacttcgag gaagtggtgg 4260
acaagggcgc ttccgcccag agcttcatcg agcggatgac caacttcgat aagaacctgc 4320
ccaacgagaa ggtgctgccc aagcacagcc tgctgtacga gtacttcacc gtgtataacg 4380
agctgaccaa agtgaaatac gtgaccgagg gaatgagaaa gcccgccttc ctgagcggcg 4440
agcagaaaaa ggccatcgtg gacctgctgt tcaagaccaa ccggaaagtg accgtgaagc 4500
agctgaaaga ggactacttc aagaaaatcg agtgcttcga ctccgtggaa atctccggcg 4560
tggaagatcg gttcaacgcc tccctgggca cataccacga tctgctgaaa attatcaagg 4620
acaaggactt cctggacaat gaggaaaacg aggacattct ggaagatatc gtgctgaccc 4680
tgacactgtt tgaggacaga gagatgatcg aggaacggct gaaaacctat gcccacctgt 4740
tcgacgacaa agtgatgaag cagctgaagc ggcggagata caccggctgg ggcaggctga 4800
gccggaagct gatcaacggc atccgggaca agcagtccgg caagacaatc ctggatttcc 4860
tgaagtccga cggcttcgcc aacagaaact tcatgcagct gatccacgac gacagcctga 4920
cctttaaaga ggacatccag aaagcccagg tgtccggcca gggcgatagc ctgcacgagc 4980
acattgccaa tctggccggc agccccgcca ttaagaaggg catcctgcag acagtgaagg 5040
tggtggacga gctcgtgaaa gtgatgggcc ggcacaagcc cgagaacatc gtgatcgaaa 5100
tggccagaga gaaccagacc acccagaagg gacagaagaa cagccgcgag agaatgaagc 5160
ggatcgaaga gggcatcaaa gagctgggca gccagatcct gaaagaacac cccgtggaaa 5220
acacccagct gcagaacgag aagctgtacc tgtactacct gcagaatggg cgggatatgt 5280
acgtggacca ggaactggac atcaaccggc tgtccgacta cgatgtggac catatcgtgc 5340
ctcagagctt tctgaaggac gactccatcg acaacaaggt gctgaccaga agcgacaaga 5400
accggggcaa gagcgacaac gtgccctccg aagaggtcgt gaagaagatg aagaactact 5460
ggcggcagct gctgaacgcc aagctgatta cccagagaaa gttcgacaat ctgaccaagg 5520
ccgagagagg cggcctgagc gaactggata aggccggctt catcaagaga cagctggtgg 5580
aaacccggca gatcacaaag cacgtggcac agatcctgga ctcccggatg aacactaagt 5640
acgacgagaa tgacaagctg atccgggaag tgaaagtgat caccctgaag tccaagctgg 5700
tgtccgattt ccggaaggat ttccagtttt acaaagtgcg cgagatcaac aactaccacc 5760
acgcccacga cgcctacctg aacgccgtcg tgggaaccgc cctgatcaaa aagtacccta 5820
agctggaaag cgagttcgtg tacggcgact acaaggtgta cgacgtgcgg aagatgatcg 5880
ccaagagcga gcaggaaatc ggcaaggcta ccgccaagta cttcttctac agcaacatca 5940
tgaacttttt caagaccgag attaccctgg ccaacggcga gatccggaag cggcctctga 6000
tcgagacaaa cggcgaaacc ggggagatcg tgtgggataa gggccgggat tttgccaccg 6060
tgcggaaagt gctgagcatg ccccaagtga atatcgtgaa aaagaccgag gtgcagacag 6120
gcggcttcag caaagagtct atcctgccca agaggaacag cgataagctg atcgccagaa 6180
agaaggactg ggaccctaag aagtacggcg gcttcgacag ccccaccgtg gcctattctg 6240
tgctggtggt ggccaaagtg gaaaagggca agtccaagaa actgaagagt gtgaaagagc 6300
tgctggggat caccatcatg gaaagaagca gcttcgagaa gaatcccatc gactttctgg 6360
aagccaaggg ctacaaagaa gtgaaaaagg acctgatcat caagctgcct aagtactccc 6420
tgttcgagct ggaaaacggc cggaagagaa tgctggcctc tgccggcgaa ctgcagaagg 6480
gaaacgaact ggccctgccc tccaaatatg tgaacttcct gtacctggcc agccactatg 6540
agaagctgaa gggctccccc gaggataatg agcagaaaca gctgtttgtg gaacagcaca 6600
agcactacct ggacgagatc atcgagcaga tcagcgagtt ctccaagaga gtgatcctgg 6660
ccgacgctaa tctggacaaa gtgctgtccg cctacaacaa gcaccgggat aagcccatca 6720
gagagcaggc cgagaatatc atccacctgt ttaccctgac caatctggga gcccctgccg 6780
ccttcaagta ctttgacacc accatcgacc ggaagaggta caccagcacc aaagaggtgc 6840
tggacgccac cctgatccac cagagcatca ccggcctgta cgagacacgg atcgacctgt 6900
ctcagctggg aggcgacaaa aggccggcgg ccacgaaaaa ggccggccag gcaaaaaaga 6960
aaaagtaagg atcctgattg atcgatagag ctcgaatttc cccgatcgtt caaacatttg 7020
gcaataaagt ttcttaagat tgaatcctgt tgccggtctt gcgatgatta tcatataatt 7080
tctgttgaat tacgttaagc atgtaataat taacatgtaa tgcatgacgt tatttatgag 7140
atgggttttt atgattagag tcccgcaatt atacatttaa tacgcgatag aaaacaaaat 7200
atagcgcgca aactaggata aattatcgcg cgcggtgtca tctatgttac tagatcggga 7260
attcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac 7320
acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac 7380
tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc 7440
tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggc tagagcagct 7500
tgccaacatg gtggagcacg acactctcgt ctactccaag aatatcaaag atacagtctc 7560
agaagaccaa agggctattg agacttttca acaaagggta atatcgggaa acctcctcgg 7620
attccattgc ccagctatct gtcacttcat caaaaggaca gtagaaaagg aaggtggcac 7680
ctacaaatgc catcattgcg ataaaggaaa ggctatcgtt caagatgcct ctgccgacag 7740
tggtcccaaa gatggacccc cacccacgag gagcatcgtg gaaaaagaag acgttccaac 7800
cacgtcttca aagcaagtgg attgatgtga taacatggtg gagcacgaca ctctcgtcta 7860
ctccaagaat atcaaagata cagtctcaga agaccaaagg gctattgaga cttttcaaca 7920
aagggtaata tcgggaaacc tcctcggatt ccattgccca gctatctgtc acttcatcaa 7980
aaggacagta gaaaaggaag gtggcaccta caaatgccat cattgcgata aaggaaaggc 8040
tatcgttcaa gatgcctctg ccgacagtgg tcccaaagat ggacccccac ccacgaggag 8100
catcgtggaa aaagaagacg ttccaaccac gtcttcaaag caagtggatt gatgtgatat 8160
ctccactgac gtaagggatg acgcacaatc ccactatcct tcgcaagacc ttcctctata 8220
taaggaagtt catttcattt ggagaggaca cgctgaaatc accagtctct ctctacaaat 8280
ctatctctct cgagctttcg cagatcccgg ggggcaatga gatatgaaaa agcctgaact 8340
caccgcgacg tctgtcgaga agtttctgat cgaaaagttc gacagcgtct ccgacctgat 8400
gcagctctcg gagggcgaag aatctcgtgc tttcagcttc gatgtaggag ggcgtggata 8460
tgtcctgcgg gtaaatagct gcgccgatgg tttctacaaa gatcgttatg tttatcggca 8520
ctttgcatcg gccgcgctcc cgattccgga agtgcttgac attggggagt ttagcgagag 8580
cctgacctat tgcatctccc gccgtgcaca gggtgtcacg ttgcaagacc tgcctgaaac 8640
cgaactgccc gctgttctac aaccggtcgc ggaggctatg gatgcgatcg ctgcggccga 8700
tcttagccag acgagcgggt tcggcccatt cggaccgcaa ggaatcggtc aatacactac 8760
atggcgtgat ttcatatgcg cgattgctga tccccatgtg tatcactggc aaactgtgat 8820
ggacgacacc gtcagtgcgt ccgtcgcgca ggctctcgat gagctgatgc tttgggccga 8880
ggactgcccc gaagtccggc acctcgtgca cgcggatttc ggctccaaca atgtcctgac 8940
ggacaatggc cgcataacag cggtcattga ctggagcgag gcgatgttcg gggattccca 9000
atacgaggtc gccaacatct tcttctggag gccgtggttg gcttgtatgg agcagcagac 9060
gcgctacttc gagcggaggc atccggagct tgcaggatcg ccacgactcc gggcgtatat 9120
gctccgcatt ggtcttgacc aactctatca gagcttggtt gacggcaatt tcgatgatgc 9180
agcttgggcg cagggtcgat gcgacgcaat cgtccgatcc ggagccggga ctgtcgggcg 9240
tacacaaatc gcccgcagaa gcgcggccgt ctggaccgat ggctgtgtag aagtactcgc 9300
cgatagtgga aaccgacgcc ccagcactcg tccgagggca aagaaataga gtagatgccg 9360
accggatctg tcgatcgaca agctcgagtt tctccataat aatgtgtgag tagttcccag 9420
ataagggaat tagggttcct atagggtttc gctcatgtgt tgagcatata agaaaccctt 9480
agtatgtatt tgtatttgta aaatacttct atcaataaaa tttctaattc ctaaaaccaa 9540
aatccagtac taaaatccag atcccccgaa ttaattcggc gttaattcag tacattaaaa 9600
acgtccgcaa tgtgttatta agttgtctaa gcgtcaattt gtttacacca caatatatcc 9660
tgccaccagc cagccaacag ctccccgacc ggcagctcgg cacaaaatca ccactcgata 9720
caggcagccc atcagtccgg gacggcgtca gcgggagagc cgttgtaagg cggcagactt 9780
tgctcatgtt accgatgcta ttcggaagaa cggcaactaa gctgccgggt ttgaaacacg 9840
gatgatctcg cggagggtag catgttgatt gtaacgatga cagagcgttg ctgcctgtga 9900
tcaccgcggt ttcaaaatcg gctccgtcga tactatgtta tacgccaact ttgaaaacaa 9960
ctttgaaaaa gctgttttct ggtatttaag gttttagaat gcaaggaaca gtgaattgga 10020
gttcgtcttg ttataattag cttcttgggg tatctttaaa tactgtagaa aagaggaagg 10080
aaataataaa tggctaaaat gagaatatca ccggaattga aaaaactgat cgaaaaatac 10140
cgctgcgtaa aagatacgga aggaatgtct cctgctaagg tatataagct ggtgggagaa 10200
aatgaaaacc tatatttaaa aatgacggac agccggtata aagggaccac ctatgatgtg 10260
gaacgggaaa aggacatgat gctatggctg gaaggaaagc tgcctgttcc aaaggtcctg 10320
cactttgaac ggcatgatgg ctggagcaat ctgctcatga gtgaggccga tggcgtcctt 10380
tgctcggaag agtatgaaga tgaacaaagc cctgaaaaga ttatcgagct gtatgcggag 10440
tgcatcaggc tctttcactc catcgacata tcggattgtc cctatacgaa tagcttagac 10500
agccgcttag ccgaattgga ttacttactg aataacgatc tggccgatgt ggattgcgaa 10560
aactgggaag aagacactcc atttaaagat ccgcgcgagc tgtatgattt tttaaagacg 10620
gaaaagcccg aagaggaact tgtcttttcc cacggcgacc tgggagacag caacatcttt 10680
gtgaaagatg gcaaagtaag tggctttatt gatcttggga gaagcggcag ggcggacaag 10740
tggtatgaca ttgccttctg cgtccggtcg atcagggagg atatcgggga agaacagtat 10800
gtcgagctat tttttgactt actggggatc aagcctgatt gggagaaaat aaaatattat 10860
attttactgg atgaattgtt ttagtaccta gaatgcatga ccaaaatccc ttaacgtgag 10920
ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct 10980
ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt 11040
tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg 11100
cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct 11160
gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc 11220
gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg 11280
tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa 11340
ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg 11400
gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg 11460
ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga 11520
tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt 11580
ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct 11640
gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga 11700
acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt 11760
ctccttacgc atctgtgcgg tatttcacac cgcatatggt gcactctcag tacaatctgc 11820
tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg 11880
ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg 11940
catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac 12000
cgtcatcacc gaaacgcgcg aggcagggtg ccttgatgtg ggcgccggcg gtcgagtggc 12060
gacggcgcgg cttgtccgcg ccctggtaga ttgcctggcc gtaggccagc catttttgag 12120
cggccagcgg ccgcgatagg ccgacgcgaa gcggcggggc gtagggagcg cagcgaccga 12180
agggtaggcg ctttttgcag ctcttcggct gtgcgctggc cagacagtta tgcacaggcc 12240
aggcgggttt taagagtttt aataagtttt aaagagtttt aggcggaaaa atcgcctttt 12300
ttctctttta tatcagtcac ttacatgtgt gaccggttcc caatgtacgg ctttgggttc 12360
ccaatgtacg ggttccggtt cccaatgtac ggctttgggt tcccaatgta cgtgctatcc 12420
acaggaaaga gtccttttcg acctttttcc cctgctaggg caatttgccc tagcatctgc 12480
tccgtacatt aggaaccggc ggatgcttcg ccctcgatca ggttgcggta gcgcatgact 12540
aggatcgggc cagcctgccc cgcctcctcc ttcaaatcgt actccggcag gtcatttgac 12600
ccgatcagct tgcgcacggt gaaacagaac ttcttgaact ctccggcgct gccactgcgt 12660
tcgtagatcg tcttgaacaa ccatctggct tctgccttgc ctgcggcgcg gcgtgccagg 12720
cggtagagaa aacggccgat gccgggatcg atcaaaaagt aatcggggtg aaccgtcagc 12780
acgtccgggt tcttgccttc tgtgatctcg cggtacatcc aatcagctag ctcgatctcg 12840
atgtactccg gccgcccggt ttcgctcttt acgatcttgt agcggctaat caaggcttca 12900
ccctcggata ccgtcaccag gcggccgttc ttggccttct tcgtacgctg catggcaacg 12960
tgcgtggtgt ttaaccgaat gcaggtttct accaggtcgt ctttctgctt tccgccatcg 13020
gctcgccggc agaacttgag tacgtccgca acgtgtggac ggaacacgcg gccgggcttg 13080
tctcccttcc cttcccggta tcggttcatg gattcggtta gatgggaaac cgccatcagt 13140
accaggtcgt aatcccacac actggccatg ccggccggcc ctgcggaaac ctctacgtgc 13200
ccgtctggaa gctcgtagcg gatcacctcg ccagctcgtc ggtcacgctt cgacagacgg 13260
aaaacggcca cgtccatgat gctgcgacta tcgcgggtgc ccacgtcata gagcatcgga 13320
acgaaaaaat ctggttgctc gtcgcccttg ggcggcttcc taatcgacgg cgcaccggct 13380
gccggcggtt gccgggattc tttgcggatt cgatcagcgg ccgcttgcca cgattcaccg 13440
gggcgtgctt ctgcctcgat gcgttgccgc tgggcggcct gcgcggcctt caacttctcc 13500
accaggtcat cacccagcgc cgcgccgatt tgtaccgggc cggatggttt gcgaccgtca 13560
cgccgattcc tcgggcttgg gggttccagt gccattgcag ggccggcaga caacccagcc 13620
gcttacgcct ggccaaccgc ccgttcctcc acacatgggg cattccacgg cgtcggtgcc 13680
tggttgttct tgattttcca tgccgcctcc tttagccgct aaaattcatc tactcattta 13740
ttcatttgct catttactct ggtagctgcg cgatgtattc agatagcagc tcggtaatgg 13800
tcttgccttg gcgtaccgcg tacatcttca gcttggtgtg atcctccgcc ggcaactgaa 13860
agttgacccg cttcatggct ggcgtgtctg ccaggctggc caacgttgca gccttgctgc 13920
tgcgtgcgct cggacggccg gcacttagcg tgtttgtgct tttgctcatt ttctctttac 13980
ctcattaact caaatgagtt ttgatttaat ttcagcggcc agcgcctgga cctcgcgggc 14040
agcgtcgccc tcgggttctg attcaagaac ggttgtgccg gcggcggcag tgcctgggta 14100
gctcacgcgc tgcgtgatac gggactcaag aatgggcagc tcgtacccgg ccagcgcctc 14160
ggcaacctca ccgccgatgc gcgtgccttt gatcgcccgc gacacgacaa aggccgcttg 14220
tagccttcca tccgtgacct caatgcgctg cttaaccagc tccaccaggt cggcggtggc 14280
ccatatgtcg taagggcttg gctgcaccgg aatcagcacg aagtcggctg ccttgatcgc 14340
ggacacagcc aagtccgccg cctggggcgc tccgtcgatc actacgaagt cgcgccggcc 14400
gatggccttc acgtcgcggt caatcgtcgg gcggtcgatg ccgacaacgg ttagcggttg 14460
atcttcccgc acggccgccc aatcgcgggc actgccctgg ggatcggaat cgactaacag 14520
aacatcggcc ccggcgagtt gcagggcgcg ggctagatgg gttgcgatgg tcgtcttgcc 14580
tgacccgcct ttctggttaa gtacagcgat aaccttcatg cgttcccctt gcgtatttgt 14640
ttatttactc atcgcatcat atacgcagcg accgcatgac gcaagctgtt ttactcaaat 14700
acacatcacc tttttagacg gcggcgctcg gtttcttcag cggccaagct ggccggccag 14760
gccgccagct tggcatcaga caaaccggcc aggatttcat gcagccgcac ggttgagacg 14820
tgcgcgggcg gctcgaacac gtacccggcc gcgatcatct ccgcctcgat ctcttcggta 14880
atgaaaaacg gttcgtcctg gccgtcctgg tgcggtttca tgcttgttcc tcttggcgtt 14940
cattctcggc ggccgccagg gcgtcggcct cggtcaatgc gtcctcacgg aaggcaccgc 15000
gccgcctggc ctcggtgggc gtcacttcct cgctgcgctc aagtgcgcgg tacagggtcg 15060
agcgatgcac gccaagcagt gcagccgcct ctttcacggt gcggccttcc tggtcgatca 15120
gctcgcgggc gtgcgcgatc tgtgccgggg tgagggtagg gcgggggcca aacttcacgc 15180
ctcgggcctt ggcggcctcg cgcccgctcc gggtgcggtc gatgattagg gaacgctcga 15240
actcggcaat gccggcgaac acggtcaaca ccatgcggcc ggccggcgtg gtggtgtcgg 15300
cccacggctc tgccaggcta cgcaggcccg cgccggcctc ctggatgcgc tcggcaatgt 15360
ccagtaggtc gcgggtgctg cgggccaggc ggtctagcct ggtcactgtc acaacgtcgc 15420
cagggcgtag gtggtcaagc atcctggcca gctccgggcg gtcgcgcctg gtgccggtga 15480
tcttctcgga aaacagcttg gtgcagccgg ccgcgtgcag ttcggcccgt tggttggtca 15540
agtcctggtc gtcggtgctg acgcgggcat agcccagcag gccagcggcg gcgctcttgt 15600
tcatggcgta atgtctccgg ttctagtcgc aagtattcta ctttatgcga ctaaaacacg 15660
cgacaagaaa acgccaggaa aagggcaggg cggcagcctg tcgcgtaact taggacttgt 15720
gcgacatgtc gttttcagaa gacggctgca ctgaacgtca gaagccgact gcactatagc 15780
agcggagggg ttggatcaaa gtactttgat cccgagggga accctgtggt tggcatgcac 15840
atacaaatgg acgaacggat aaaccttttc acgccctttt aaatatccgt tattctaa 15898
<210> 5
<211> 96
<212> DNA
<213> Artificial sequence
<400> 5
tacattgagg agcatggcac gttttagagc tagaaatagc aagttaaaat aaggctagtc 60
cgttatcaac ttgaaaaagt ggcaccgagt cggtgc 96
<210> 6
<211> 20
<212> RNA
<213> Artificial sequence
<400> 6
uacauugagg agcauggcac 20
Claims (10)
1. a kind of protein, for as follows (a1) or (a2) or (a3) or (a4):
(a1) protein shown in sequence 1 in sequence table；
(a2) fusion protein obtained in N-terminal or/and C-terminal the connection label of (a1) described protein；
(a3) what is obtained (a1) by the substitution and/or deletion and/or addition of one or several amino acid residues sends out with plant
Educate relevant protein；
(a4) there is 98% or more identity and protein relevant with development of plants from rice and to (a1).
2. encoding the nucleic acid molecules of protein described in claim 1.
3. nucleic acid molecules as claimed in claim 2, it is characterised in that: the nucleic acid molecules are following (b1) or (b2) or (b3)
Or (b4):
(b1) code area DNA molecular as shown in sequence 2 in sequence table；
(b2) DNA molecular shown in sequence 3 in sequence table；
(b3) there is the DNA molecular of 95% or more identity and code for said proteins from rice and with (b1) or (b2)；
(b4) DNA under strict conditions with (b1) or (b2) nucleotide sequence hybridization limited and code for said proteins divides
Son.
4. expression cassette, recombinant vector or recombinant microorganism containing nucleic acid molecules described in Claims 2 or 3.
5. the application of protein described in claim 1, for as follows (c1), (c2), (c3) or (c4):
(c1) regulate and control the plant height character of plant；
(c2) regulate and control the fringe character of plant；
(c3) regulate and control the seed grain length character of plant；
(c4) regulate and control the wide character of seed grain of plant.
6. the application of nucleic acid molecules described in Claims 2 or 3, for as follows (d1), (d2), (d3) or (d4):
(d1) genetically modified plants that plant height character changes are cultivated；
(d2) genetically modified plants that fringe character changes are cultivated；
(d3) genetically modified plants that seed grain length character changes are cultivated；
(d4) genetically modified plants that the wide character of seed grain changes are cultivated.
7. inhibit the substance of protein described in claim 1 in the application of plant breeding, the target of the breeding be following (e1),
(e2), (e3) or (e4):
(e1) plant that plant height increases is cultivated；
(e2) plant that fringe increases is cultivated；
(e3) the increased plant of seed grain length is cultivated；
(e4) the wide reduced plant of seed grain is cultivated.
8. inhibit the application of the substance of nucleic acid molecules described in Claims 2 or 3, for as follows (f1), (f2), (f3) or (f4):
(f1) genetically modified plants that plant height increases are cultivated；
(f2) genetically modified plants that fringe increases are cultivated；
(f3) the increased genetically modified plants of seed grain length are cultivated；
(f4) the wide reduced genetically modified plants of seed grain are cultivated.
9. a kind of method of prepare transgenosis plant includes the following steps: to import inhibition Claims 2 or 3 in recipient plant
The substance of the nucleic acid molecules expression, obtains genetically modified plants；Compared with the recipient plant, the transgenic plant has such as
At least one of lower four kinds of phenotypes: plant height increases, and fringe increases, and seed grain length increases, the wide reduction of seed grain.
10. a kind of plant breeding method includes the following steps: the content for reducing protein described in claim 1 in purpose plant
And/or activity, to make purpose plant that at least one of following four kinds of phenotypic alternations occur: plant height increases, and fringe increases, seed
Grain length increases, the wide reduction of seed grain.
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| CN201910462534.9A CN110183523B (en) | 2019-05-30 | 2019-05-30 | OsMYB36 protein and coding gene and application thereof |
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| CN114438119A (en) * | 2022-02-16 | 2022-05-06 | 广西大学 | Preparation method and application of labeled polygene mutant rice plant |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN111205358A (en) * | 2020-03-04 | 2020-05-29 | 中国农业科学院生物技术研究所 | Os494 protein and coding gene and application thereof |
| CN111217897A (en) * | 2020-03-04 | 2020-06-02 | 中国农业科学院生物技术研究所 | Os516 protein and coding gene and application thereof |
| CN111205358B (en) * | 2020-03-04 | 2021-03-23 | 中国农业科学院生物技术研究所 | Os494 protein and coding gene and application thereof |
| CN111217897B (en) * | 2020-03-04 | 2021-06-25 | 中国农业科学院生物技术研究所 | Os516 protein and coding gene and application thereof |
| CN114805509B (en) * | 2022-01-20 | 2023-10-13 | 上海交通大学 | Application of protein OsGATA6 in regulation of plant flowering time, spike shape, grain shape and thousand grain weight |
| CN114438119A (en) * | 2022-02-16 | 2022-05-06 | 广西大学 | Preparation method and application of labeled polygene mutant rice plant |
| CN114438119B (en) * | 2022-02-16 | 2023-10-13 | 广西大学 | Preparation method and application of multi-gene mutant rice plant carrying markers |
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|---|---|
| CN110183523B (en) | 2021-02-09 |
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