CN108753781A - The application of phosphate transporter gene GmPT5 promoters - Google Patents
The application of phosphate transporter gene GmPT5 promoters Download PDFInfo
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Abstract
The present invention provides phosphate transporter genesGmPT5The application of promoter, it is specifically disclosed to beGmPT5Promoter is in driving expression of the legume gene in root nodule and the application in terms of responding low-phosphor.Pass through promoter region piecewise analysis, Ke LongliaoGmPT5Different length promoter, and demonstrateGmPT57 different lengths promoter section（Including 5 ' UTR）Expression of the gus protein in root nodule can be driven, the promoter section not comprising 5 ' UTR cannot then drive expression of the gus protein in root nodule, wherein there is expression of 1 section driving gus protein in root nodule to have response to low-phosphorous.
Description
Technical field
The present invention relates to the new opplications of gene, and in particular to phosphate transporter geneGmPT5The application of promoter belongs to raw
Object technical field.
Background technology
Phosphorus is one of macronutrient necessary to growth and development of plants.Phos by cell membrane enter cell be by
Multiple phosphate transporter genes of same or different families are responsible for, and the phosphate transporter gene being currently known is by sequence and function phase
It is divided into five large families like propertyPht1、Pht2、Pht3、Pho1WithPho2（Yang Cunyi etc., 2006）.Due toPht1Family gene master
It is responsible for absorption of the root system to soil phophorus, therefore as the plant phosphate transporter gene for studying the most deep.Pht1Phosphorus turns
First member for transporting protein family gene is from yeast（Saccharomyces eerevisae）In it is isolated（Bun-Ya
et al, 1991）.Utilize sequence conservation to analyze, respectively from arabidopsis in recent years（Muchhal et al, 1996）, potato
（Leggewie et al, 1997）, tobacco（Mitsukawa et al, 1997）, clover（Liu et al, 1998）, it is big
Wheat（Smith et al, 1999）, Lupinus albus（Liu et al, 2001）, rice（Paszkowski et al, 2002）,
Soybean（Qin et al, 2012）The many phosphate transporter genes for belonging to Pht1 families are cloned into equal higher plants.
2012, Qin et al. had reported soybeanPht1The phosphorus transporter Protein G mPT5 of family's high-affinity is in root nodule phosphorus acquisition process
Function.GmPT5It is primarily targeted for the fibrovascular system of soybean root and root nodule, phosphorus is taken part in and turns from soybean root system to root nodule
Fortune；33The experiment of P isotope situ absorptions further demonstrates, and excess or interferes effect gene phosphorus from root to the tired of root nodule
Product, showsGmPT5It is the key gene for regulating and controlling root nodule phosphorus dynamic equilibrium under low-phosphorus stress, ensure that root nodule is to phosphorus under low-phosphorus stress
High requirements（Qin et al., 2012）.
Currently,Pht1The research of phosphorus transporter protein family gene focuses primarily upon on transcriptional level, the promoter of gene
The tissue specificity of gene expression is determined with some transcription factor interactions and to the response of environmental change（Berman et al.,
2002; Le Hir et al., 2003; Schallau et al., 2008）.Promoter sequence is located at the upstream of gene, certainly
Determine transcription initiation site and frequency.Most of promoter not only contains TATA-box, and there is also the cis actings of some specificity
Element, such as low-phosphorous response element P1BS, root nodule related elements NODCON1GM, NODCON2GM, mycorhiza related elements MYCS, PB1S
（Daram et al., 1998; Liu et al., 1998; Rubio et al., 2001; Schachtman & Shin,
2007; Chen et al., 2011）.These cis elements are combined by the transcription regulatory factor of upstream, for controlling gene
Expression has great importance.The regulatory molecule mechanism of research promoter helps to understand complicated gene regulatory network.Currently,
Research about promoter is concentrated mainly on the identification of its clone and crucial cis-acting elements.
Invention content
The object of the present invention is to provide phosphate transporter genesGmPT5The application of promoter.
To achieve the above object, the present invention uses following technical scheme：
Phosphate transporter geneGmPT5Promoter is in terms of driving expression and responding low-phosphor of the legume gene in root nodule
Application.
The phosphate transporter geneGmPT5Promoter sequence is as shown in SEQ ID NO.1.
The phosphate transporter geneGmPT5 containsThe promoter section of 7 different lengths, each sector sequence such as SEQ ID
Shown in NO.2-8.
Inventor according toGmPT5Promoter sequence design 8 different length promoter section, pass through fusionGUS
Reporter gene utilizes expression of the transgenosis hair root Rhizobium Inoculation observation gus protein in root nodule, the results showed thatGmPT5
7 different lengths promoter section（Including 5 ' UTR）Expression of the gus protein in root nodule can be driven, 5 ' UTR are not included
Promoter section cannot then drive expression of the gus protein in root nodule, wherein there is 1 section to drive gus protein in root nodule
Expression have response to low-phosphorous.
The invention has the beneficial effects that：This research drives gene specific in root the legume including soybean
It is expressed in tumor, especially provides alternate promoters resource to the gene of low-phosphorous response.
Description of the drawings
Fig. 1 is the ideograph of 8 promoter sections.
Fig. 2 is root nodule GUS colored graphs.
Fig. 3 is root nodule slice map.
Fig. 4 is that GUS quantitatively schemes.
Specific implementation mode
Specific introduce is made to the present invention below in conjunction with the drawings and specific embodiments.
Embodiment 1,GmPT5The clone of promoter
It is downloaded in the soybean genome database announcedGmPT5Promoter sequence, use TSSP-TCM online softwares
（http://mendel.cs.rhul.ac.uk/mendel.phptopic=fgen）（Shahmuradov etc. 2005）To sequencing
As a result it is analyzed, predicts transcription initiation site, use NCBI online softwares（http://www.ncbi.nlm.nih.gov/）
Sequencing result is analyzed, conjunctive use PLACE（http://www.dna.affrc.go.jp/PLACE/）（Higo etc.
1998, 1999）PlantCARE （http://bioinfor-matics.psb.ugent.be/webtools/plantcare/
html/）（Rom-bauts etc. 1999;Lescot etc. 2002）Online software analyzes its transcriptional regulatory element.This is opened
The different sections of mover withGUSGene closes construction of expression vector.
The leaf for taking soybean line HN89, using CTAB methods extraction total DNA as cloneGmPT5The mould of promoter fragment
Plate.According to soybeanGmPT5Gene（Glyma10g04230.1）5' flanking sequence design primers, primer are as follows：
It is separately added into NcoI and XbaI enzyme cutting site and protection base at the ends 5' of upstream and downstream primer.50 μ L PCR reaction system packets
It includes:1 μ L Soybean genomic DNAs, 5 μ L 10 × LA PCR buffer solutions, 8 μ L（2.5 mmol/L）DNTP, 10 μm/μ L it is upper,
Each 2 μ L of downstream primer, 0.5 μ L LA Taq archaeal dna polymerases（5 U/μL ）,31.5 μL ddH2O.PCR reaction cycle conditions
For 94 DEG C of 2 min of pre-degeneration;94 DEG C of denaturation 30 s, 53 DEG C of annealing 30 s, 72 DEG C of 3.1 min of extension carry out 30 altogether
Cycle;10 min of last 72 DEG C of extensions.PCR product carries out electrophoresis with 1.0% Ago-Gel, then cuts the mesh of 3092bp
Band, use gel reclaims kit recycling, purifying target fragment.Purified product is connect with pMD18-T carriers, then is used
DH10B competent cells are converted, even spread to benzyl containing ammonia（100 mg/L）LB tablets on, trained in 37 DEG C of baking ovens
12 ~ 13 h are supported, picking monoclonal colonies carry out PCR detections, obtain positive colony, are named as pro PT5- T, positive colony committee
Support Beijing AudioCodes biotechnology Services Co., Ltd is sequenced.
With pro PT5- T recombinant plasmids are that template carries out PCR amplification, respectively obtain 3092 (SEQ ID NO.1), 2459
(SEQ ID NO.2)、1449 (SEQ ID NO.3)、1215 (SEQ ID NO.4)、1146 (SEQ ID NO.5)、401
(SEQ ID NO.6),137 (SEQ ID NO.7),1175 (SEQ ID NO.8)（The startup of 5 ' UTR and introne are not included
Sub-piece）This 8 segments are replaced the 35S promoter in plant expression vector pCAMBIA3301, make each by 8 segments
Section respectively in carrierGUSReporter gene fusion, to build each promoter fragment carrier.pro3092::GUS、
pro2459::GUS、pro1449::GUS、pro1215::GUS 、pro1146::GUS 、pro401::GUS、pro137::GUS
And pro1175::GUS8 plant expression vectors.
Fig. 1 is the ideograph of 8 promoter sections.
The result shows that：WithGmPT5Promoter sequences of 3092 bp as the gene before translation initiation site ATG, analysis hair
Existing, which includesGmPT5The sequence of 1325 bp of the ends gene 5 ' UTR.It willGmPT5Promoter fragment（3092 bp）Pass through
TSSP-TCM（Shahmuradov et al., 2005）Its transcription initiation site of on-line prediction is located at the 1305bp of the upstreams ATG,
TATA boxes are predicted in the position of transcription initiation upstream -24 to -30, further by promoter fragment in plant cis regulatory DNA members
The prediction of controlling element is carried out in part analytical database, interpretation of result is found,GmPT5Exist in promoter with root nodule, corolla etc.
Position is expressed and low-phosphorous relevant critical elements.GmPT5It, may be with the relevant regulation and control member of root nodule specifically expressing in promoter sequence
Part（AAAGAT）With（CTCTT）There are 4 and 2 respectively, develops relevant element with flower and corolla（AGAAA）There are 4, with root
The hair relevant element of specifically expressing（KCACGW）There is 1；Only find the binding sequence of 1 PHR1 related with scarce phosphorus signal
（GNATATNC）；Additionally, it is observed that with low temperature and the relevant controlling element of sucrose, and respectively have 1 in the gene promoter sequence
It is a.
Embodiment 2,GmPT5Promoter section driving gus protein expressed in root nodule
The transgene expression vector pro built PT5After-T converts Agrobacterium K599, turned by agriculture bacillus mediated hypocotyl
Change method（Kereszt etc. 2007）After obtaining transgenic hairy root, main root is cut, retains the hairy come out from callus director
Then hairy root is immersed in rhizobium bacterium solution and moves into water planting after 1 hour by root, mill water culture nutrient solution is all low nitrogen（500μmol/
L）, it is low-phosphorous（5 μmol/L）, high phosphorus（250 μmol/L）Processing, root growth after a week, when beginning with root nodule and occurring, take one plant
Upper all hair roots of callus（General 5 or so）Lateral root carry out GUS dyeing, remove non-transgenic hairy root, after inoculation
Root nodule is won within 15 days, 30 days respectively and carries out the active tissue chemical analysis of GUS, determines the gene in the expression portion of different phase
Position.
With reference to Raghothama etc.（1999）Method in article, sample are first cleaned twice with 0.2 M phosphate buffers
Afterwards, after GUS dye liquors being added, 37 DEG C of baking ovens are put into, sample outwells dye liquor after there is obvious blue, impregnates 20-30 with 50% alcohol
Stereomicroscope is used after minute（LEICA DFC420, Germany）Observe bulk dyeing situation.
The tissue positioning of gene expression can be observed further by paraffin section.The specific method is as follows：
(1) by the material cut FAA fixers（50% or 70% alcohol, 90 mL+ glacial acetic acid, 5 mL+ formaldehyde, 5 mL）It vacuumizes
It is fixed to stay overnight；
(2) 1.5 hours are dehydrated successively with 75%, 85%, 95% alcohol, then 100% alcohol continues dehydration three after fixed
It is secondary, 1 hour every time；
(3) material transparent is carried out with+1/2 absolute alcohol of 1/2 dimethylbenzene and pure dimethylbenzene within 1.5 hours every time, then after dehydration
Inside dimethylbenzene plus 40 DEG C of paraffin is stayed overnight；
(4) and then with paraffin refined wax wax being carried out at 60 DEG C, changing pure wax within every 1.5 hours, this step is repeated 3 times；
(5) material is embedded in paraffin refined wax；
(6) with semi-automatic slicer（CUT 5062）Slice, thickness are 15 μm.
(7) light microscope is used（LEICA DM5000B）Observation is carried out to take pictures.
The result shows that：The bright different length of bulk dyeing chartGmPT5Promoter（In addition to 1175bp promoter sections）?
It can drivingGUSGene is expressed in the vascular tissue of root and root nodule, negative control（The arabidopsis reportedAtPT4Promoter
Section）Without gus protein expression activity；Positive control（CaMV35Spro::GUS）There is gus protein expression to live in root and root nodule
Property.Section not comprising 5 ' UTR is unable to section gus protein and is expressed in root nodule, illustrates the 5 ' UTR of GmPT5 to driving gene
Expression is essential.137bp sections promoter has response to low-phosphorous, may contain low-phosphorous response element, referring to Fig. 2.
As can be drawn from Figure 3, in low-phosphorous processing, root nodule slice display of the Rhizobium Inoculation after 7 days：The startup of 401bp
Son can drive gus protein in the expression of the vascular tissue of the intersection and root of root and root nodule.Root nodule of the Rhizobium Inoculation after 15 days
Slice display：Expression of the promoter driving gus protein of 137bp in the vascular tissue of root and root nodule.Rhizobium Inoculation 20 days
Root nodule slice display afterwards：The promoter of 3092bp can drive gus gene to be expressed in the vascular tissue of root nodule and root system, and
Root nodule infects the expression in area's cell.Root nodule slice display of the Rhizobium Inoculation after 30 days：The promoter of 1449bp, 2496bp are driven
Dynamic gus protein is expressed in the vascular tissue of root nodule and root system, and the section 1175bp not comprising 5 ' UTR cannot drive gus protein
It is expressed in root and root nodule.
Embodiment 3,GmPT5Promoter section driving gus gene expressed in root nodule
Root and root nodule are harvested respectively, and RT-PCR experiments are carried out after extraction RNA reversions, it is rightGmPT5Gene andGUSThe expression mould of gene
Formula is analyzed, and quantification PCR primer is as follows：
GmPT5-F: GAACACTTTCAGGGCAACTC；
GmPT5-R: GTCATCACAGTCTTTGCATCG；
GUS-F: TGTTGATGTGCAGGTATCACC；
GUS-R: GTCACCACGGTGATATCGTC；
TefS1-F: TGCAAAGGAGGCTGCTAACT；
TefS1-R: CAGCATCACCGTTCTTCAAA；
The result shows that：In low-phosphorous processing, Rhizobium Inoculation is after 30 days,GmPT5The promoter section driving of geneGUSGene exists
Expression quantity in root and root nodule is below the expression quantity in positive control 3301,137bp, 2496bp, 3092bp drivingGUS
Expression quantity of the gene in root and root nodule is all remarkably higher than to be driven not comprising 1175bpGUSThe expression quantity of gene, referring to Fig. 4.
It should be noted that the invention is not limited in any way for above-described embodiment, it is all to use equivalent replacement or equivalent change
The technical solution that the mode changed is obtained, all falls in protection scope of the present invention.
SEQUENCE LISTING
<110>University Of Agriculture and Forestry In Fujian
<120>The application of phosphate transporter gene GmPT5 promoters
<130> 30
<160> 30
<170> PatentIn version 3.3
<210> 1
<211> 3092
<212> DNA
<213>Artificial sequence 3090
<400> 1
ccaaatccag tccacggttt gtcttctatg ctcttctaca gtggaaactg atgtgcattt 60
aattttaaat tgtcgacgga tgagaccaat ttggaattta agtctgcagt ggttaaacga 120
aatgtatgtt atcccaggga ctaccattga gcattttatg tcacaaagtg gggtggcagg 180
ctgcaaggca agagtaatga aacaatgtat atgggcatct atcagctaga aacatctggc 240
gaaatagaaa ccaaattctg tttaatcaat ccgcgtggaa acttcagaag atttttgata 300
atgtgaagtt tttcttatag ctatggctta aaaatatgtt ttcctcgagc acaagtacct 360
tcacacagtg gcaagtagat atatctgtct cattacaaca actaatgtag ggagatttat 420
tgttggtagc catgcaggaa acacagtcct aaatattatg ctgctattaa tgatggtatg 480
gggcatgaat taatacatgg aagtcggaac acatagtgaa tagcaagtgg ccctctagac 540
tcggtaattg aatagtgcac taccatgcgt tgaattggtg gggcaacttt aatttttttt 600
ttttttcctt ttgtaaataa cacaggcata tatgctgggt aggaggagtg cattcgtttt 660
ggtcagtcaa atgttgggta acacttggtt agttttattt ggggtggtgt gtgctctaaa 720
tagaatgtaa ccaagtttgt aacagggcaa ctacctctaa tgttgtttct gtgattaata 780
cataatttaa tttgcttata aaaaaaaagt cctgtagagt ctaacagtct aaaatgtttg 840
atcaaacatt accctgtaga tatttctggt taactttgga ctgatataca aaaagacatg 900
tccatcaact gttacctttt gatggaaatt aatattgtta ccacgccggc cggcatcttt 960
ggtaattcga agcattaaca ggactttgtc tatactttaa attgcagtag tatctaaagt 1020
gattgtataa ttttgacata taaaaaacta actttcccgt gattcaccag aaaaagttag 1080
tacgatattt ttggcattta tcatgtaaag acagtatatg ctaaacatca tgaattgaat 1140
ttcacgaatg caaatcttaa aaatattgaa agggtgtaac taatttaagt agctgacatt 1200
tataaccgat ttaccttcct ctatcatata ccaaaggggt tacactcgta accatttgta 1260
taaatcctta cgttaagcaa accacaaact tgaattcaaa attatctcta acttgtttct 1320
cttggattct ttggagtggt cttcatgcta ctccagattt caatttactt atgctcttct 1380
gcactacttt atggtgtttg aaggcctaca tcaacgtccc tgaaccgcgc ggcccacggc 1440
accgtgtggg ggtatgtatg gggcacatga tactgtcgtt gtttgctttt ctaattggct 1500
taaatattat ttgttttcat gtaagttcat attgtttcaa ttttggtctt gtaaaaattt 1560
tggttgtctc cgtaaattta tgttttttaa ttttggtcac gatgaaatat tttttttatt 1620
ttgtttctat aaatttatat ttttttaatt ttagtctgac acaaacttat aagaattata 1680
aataaaatta aaatcttaaa ggactaaaat aaaataaata aaacacacaa atttataagg 1740
attaaaaata aaaaaaaaat cacaggaatt agaaatagaa caacatcaac ttaccaatcc 1800
taaattaaaa aaatttacaa gggaccaaaa ataaaaaagg gctaacctaa catgaccaaa 1860
acatatttaa gctttctaat ttctaatttt tcatatagtc cacattataa gtgagattta 1920
gattttgttt tgtttgctat tctatttaaa ggatcatttt cagttgcatg tgcaacttgt 1980
acgtcactca taaatagaaa acgatatctg acccattcgg tatttttttt ttttacccaa 2040
gattaaaagg ctaccctatg atacaaattt ttgttttcac tcgcttataa aacctaaagt 2100
tttttgtacc ctctgttgca ttaaaggtga gatctaacaa attattatta tgaaatcaac 2160
ctaacagaat gtaggaaatt ttggggccgc aaccgatatg gctttttttt ccacgtttta 2220
ttgtttgtat gcctctcctg ttatttttac tccttttaaa gcaatgtcct ttaaaagaat 2280
atcctttcca tattatacta tctatttggt aatcactcat ttttaaaaat aggaagataa 2340
catctcaaag gttccctgct attgagtcaa gtaactgacc ttgtaaagtt tatccaaaaa 2400
tgaaatatta cttcaagagt tttttttttt ttatatcaag taatggcaca ttcaatactt 2460
atgttatctt tccttaccaa ctttattggc caagatttgg aagaatgaat tatttttatt 2520
acattgggat ttcatagtta tttgcctata tataagtgta acaacttgcc ttttttgtta 2580
gactgattaa tttagtcatc agatatgcac aattgatcat gtgattcaaa ttccattgga 2640
aacaagttga gaattcattt gtactaaatg agataattaa atcaatagta gtttgatgag 2700
aacatttcac aaagagagtg gatggaggac atggtacaga actaatcaaa ggcagagaca 2760
aagaaagagg tacgacaaca tgcccacatc tctgcacctt gcccaccttt ttttttgata 2820
tacactttat ctctctccct tttcctggtg atctttgtgg aggtttctct aaattgtctt 2880
tactttttgt ttctttccaa gttgtgtgga atagttgcac tttaagattt ctgcaattga 2940
agtgtttatc ttctattata ttattcttct tatacatcac caaatgtagt gcctgtttgt 3000
cttgcagctt catcatcatt ttcaacaaat caatatgatg gtggtgatgg gatagaggat 3060
tagccagaaa agtaaatgga gcactcatca ag 3092
<210> 2
<211> 2480
<212> DNA
<213>Artificial sequence 2496
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gcctactgct gcctgtgaat tggccacggc agggcattct tcaaaacacc gcaaaattcc 60
cttgtatttt gagtttttaa gttttaatct ttaacatgca ttttgttaca agttaaaatt 120
tattaaaaac tacttcatct gttcttattt ataagataaa tttattaata taaactaaat 180
cagttttttt ttataatatt gatagaaaca gagggagtat aaaatcatta tagttaaact 240
ttataaaatc ataaatgaaa ctcattaaac aaagagtaaa gattcacaat tttatgattt 300
ttaataaatt ttaataaata acaaaaaatt tatttaaaag cctttattca tctaaaatca 360
ttaaagtgta agttactaat atttctctta cattataaga tcttttttca agttgtcttt 420
tgcattaaat attttttaat cgaaatatct ttaattactt tacaatgaat gttataaatt 480
aaaaagataa atgcattatt tgtcttataa ggattaaaga ataagactag catataatca 540
ttaattagat aatgaataat tgagtgaaaa gaaaaacatg agtctcaata attttaaggg 600
taattttgaa aagataatag aaattgctaa aatatttaat gttactaaaa attaatcaat 660
tttcttaaaa aaatatgaat tagtgaaaag atcttatact tagagatcac gagggtatca 720
tctaattaac ttgaatattt ttttggtata taatagtgac gatgataaaa acctctgttg 780
tatttgaaaa catataaaca aaatgtctct tccaatcctt aatcagcttg caaacttcaa 840
ttatggcacg cacattagta cgttacacta tgttattagt gagttggagt gtaggaatta 900
ttattggaac acaccgaaat aaacgtaaaa atactatgta gtggtgaatg caaaaaaaaa 960
aaaaaaaatt ggcaaagaaa ttaattagag agttaataat atccaacgag caattaagga 1020
aaaaggcatc gtaggagcac aatagctgtt gaatggagcg aagagagcgc gaaaagctaa 1080
ggtatatgct tgtcgaatat tttggacaag tgtggttttc cttttcccat gttctgttat 1140
attctataaa tcattggcca ctctccattc acctcttagt gtgtaaccaa ttcttctctt 1200
ttgctttctc tctccaaatt catcacaact tctctcgtac ctctgcaggt cagattcata 1260
ataatattct ttttgtcctt tttgttgcat cgaatccttc tatatattgt cgcatcccca 1320
tgcatgcatt atgtcactac gtctactctt gatgtttctt ttatatccca tttcatatca 1380
tgttttttca tgaatttacc tgtttatgtt ctgttttact cttatcctaa tttaatagat 1440
gtgccattta gacgtagcat acatgactca gcaagacgat ttctgaattg atttgcagat 1500
attggcaggg atttaatgat tgtctttgca tgaaaattga aaactgaaag caaatatatt 1560
tacattacca ttaattttgt gtgtatgtca tttgtttttt gagttcagat cgtgatatct 1620
tttgcatttt ctcgtggcag cagcgctagt ttatcatata tgaattcacc tgtacctgtt 1680
tgttctctta tgtcctggtc ttctttctaa aggacatatt tgttcgatta caatttttcc 1740
caatgaattt tggtacatct gaagcatact ttttcatttt ttctccctca tacgtttccc 1800
cttcggaggc aatcctattt gaggtactga tgggcactca tagtgggcat ctctcccagc 1860
gaggattcga accttggttc gtcacgttgt ggagaactag tatctcgttt tgttaaacgt 1920
attggataca tattaaaata aatttgttca gaaatattag tatgcatctt gagtacgact 1980
ttaattagat aattgggtat agtttggtat atcttaaata ctagtttgat tgcccctatc 2040
ttatattctt cttttatgtt attaaaaaat tgaattagca tagctttaat tttttatgta 2100
actaccgtgc tcgctcaact tggttggaaa ggccttccag ttttccaata taaggtaaca 2160
catggtgctg tagctggatt actctgcatg tattgtttaa catgctttta cctttcttat 2220
gttgtgtaac tagaattgaa attctatctg tacataattg attatctcgc agcctgattg 2280
agttagtatt ttttatggtt ctagtactgt tcatggtcat catatggcag tcactaatga 2340
aaaatgaccc tgcatatgca tattggtgct atgtttttga acttttagca caatactttg 2400
taatctgttg catccacttt tgcagagaag taaacatata ggagtgagag ggagggagag 2460
agaaggaaag cttggtagcg 2480
<210> 3
<211> 1433
<212> DNA
<213>Artificial sequence 1449
<400> 3
gttgaatgga gcgaagagag cgcgaaaagc taaggtatat gcttgtcgaa tattttggac 60
aagtgtggtt ttccttttcc catgttctgt tatattctat aaatcattgg ccactctcca 120
ttcacctctt agtgtgtaac caattcttct cttttgcttt ctctctccaa attcatcaca 180
acttctctcg tacctctgca ggtcagattc ataataatat tctttttgtc ctttttgttg 240
catcgaatcc ttctatatat tgtcgcatcc ccatgcatgc attatgtcac tacgtctact 300
cttgatgttt cttttatatc ccatttcata tcatgttttt tcatgaattt acctgtttat 360
gttctgtttt actcttatcc taatttaata gatgtgccat ttagacgtag catacatgac 420
tcagcaagac gatttctgaa ttgatttgca gatattggca gggatttaat gattgtcttt 480
gcatgaaaat tgaaaactga aagcaaatat atttacatta ccattaattt tgtgtgtatg 540
tcatttgttt tttgagttca gatcgtgata tcttttgcat tttctcgtgg cagcagcgct 600
agtttatcat atatgaattc acctgtacct gtttgttctc ttatgtcctg gtcttctttc 660
taaaggacat atttgttcga ttacaatttt tcccaatgaa ttttggtaca tctgaagcat 720
actttttcat tttttctccc tcatacgttt ccccttcgga ggcaatccta tttgaggtac 780
tgatgggcac tcatagtggg catctctccc agcgaggatt cgaaccttgg ttcgtcacgt 840
tgtggagaac tagtatctcg ttttgttaaa cgtattggat acatattaaa ataaatttgt 900
tcagaaatat tagtatgcat cttgagtacg actttaatta gataattggg tatagtttgg 960
tatatcttaa atactagttt gattgcccct atcttatatt cttcttttat gttattaaaa 1020
aattgaatta gcatagcttt aattttttat gtaactaccg tgctcgctca acttggttgg 1080
aaaggccttc cagttttcca atataaggta acacatggtg ctgtagctgg attactctgc 1140
atgtattgtt taacatgctt ttacctttct tatgttgtgt aactagaatt gaaattctat 1200
ctgtacataa ttgattatct cgcagcctga ttgagttagt attttttatg gttctagtac 1260
tgttcatggt catcatatgg cagtcactaa tgaaaaatga ccctgcatat gcatattggt 1320
gctatgtttt tgaactttta gcacaatact ttgtaatctg ttgcatccac ttttgcagag 1380
aagtaaacat ataggagtga gagggaggga gagagaagga aagcttggta gcg 1433
<210> 4
<211> 1199
<212> DNA
<213>Artificial sequence 1215
<400> 4
ttgttgcatc gaatccttct atatattgtc gcatccccat gcatgcatta tgtcactacg 60
tctactcttg atgtttcttt tatatcccat ttcatatcat gttttttcat gaatttacct 120
gtttatgttc tgttttactc ttatcctaat ttaatagatg tgccatttag acgtagcata 180
catgactcag caagacgatt tctgaattga tttgcagata ttggcaggga tttaatgatt 240
gtctttgcat gaaaattgaa aactgaaagc aaatatattt acattaccat taattttgtg 300
tgtatgtcat ttgttttttg agttcagatc gtgatatctt ttgcattttc tcgtggcagc 360
agcgctagtt tatcatatat gaattcacct gtacctgttt gttctcttat gtcctggtct 420
tctttctaaa ggacatattt gttcgattac aatttttccc aatgaatttt ggtacatctg 480
aagcatactt tttcattttt tctccctcat acgtttcccc ttcggaggca atcctatttg 540
aggtactgat gggcactcat agtgggcatc tctcccagcg aggattcgaa ccttggttcg 600
tcacgttgtg gagaactagt atctcgtttt gttaaacgta ttggatacat attaaaataa 660
atttgttcag aaatattagt atgcatcttg agtacgactt taattagata attgggtata 720
gtttggtata tcttaaatac tagtttgatt gcccctatct tatattcttc ttttatgtta 780
ttaaaaaatt gaattagcat agctttaatt ttttatgtaa ctaccgtgct cgctcaactt 840
ggttggaaag gccttccagt tttccaatat aaggtaacac atggtgctgt agctggatta 900
ctctgcatgt attgtttaac atgcttttac ctttcttatg ttgtgtaact agaattgaaa 960
ttctatctgt acataattga ttatctcgca gcctgattga gttagtattt tttatggttc 1020
tagtactgtt catggtcatc atatggcagt cactaatgaa aaatgaccct gcatatgcat 1080
attggtgcta tgtttttgaa cttttagcac aatactttgt aatctgttgc atccactttt 1140
gcagagaagt aaacatatag gagtgagagg gagggagaga gaaggaaagc ttggtagcg 1199
<210> 5
<211> 1130
<212> DNA
<213>Artificial sequence 1146
<400> 5
gatgtttctt ttatatccca tttcatatca tgttttttca tgaatttacc tgtttatgtt 60
ctgttttact cttatcctaa tttaatagat gtgccattta gacgtagcat acatgactca 120
gcaagacgat ttctgaattg atttgcagat attggcaggg atttaatgat tgtctttgca 180
tgaaaattga aaactgaaag caaatatatt tacattacca ttaattttgt gtgtatgtca 240
tttgtttttt gagttcagat cgtgatatct tttgcatttt ctcgtggcag cagcgctagt 300
ttatcatata tgaattcacc tgtacctgtt tgttctctta tgtcctggtc ttctttctaa 360
aggacatatt tgttcgatta caatttttcc caatgaattt tggtacatct gaagcatact 420
ttttcatttt ttctccctca tacgtttccc cttcggaggc aatcctattt gaggtactga 480
tgggcactca tagtgggcat ctctcccagc gaggattcga accttggttc gtcacgttgt 540
ggagaactag tatctcgttt tgttaaacgt attggataca tattaaaata aatttgttca 600
gaaatattag tatgcatctt gagtacgact ttaattagat aattgggtat agtttggtat 660
atcttaaata ctagtttgat tgcccctatc ttatattctt cttttatgtt attaaaaaat 720
tgaattagca tagctttaat tttttatgta actaccgtgc tcgctcaact tggttggaaa 780
ggccttccag ttttccaata taaggtaaca catggtgctg tagctggatt actctgcatg 840
tattgtttaa catgctttta cctttcttat gttgtgtaac tagaattgaa attctatctg 900
tacataattg attatctcgc agcctgattg agttagtatt ttttatggtt ctagtactgt 960
tcatggtcat catatggcag tcactaatga aaaatgaccc tgcatatgca tattggtgct 1020
atgtttttga acttttagca caatactttg taatctgttg catccacttt tgcagagaag 1080
taaacatata ggagtgagag ggagggagag agaaggaaag cttggtagcg 1130
<210> 6
<211> 385
<212> DNA
<213>Artificial sequence 401
<400> 6
atgtaactac cgtgctcgct caacttggtt ggaaaggcct tccagttttc caatataagg 60
taacacatgg tgctgtagct ggattactct gcatgtattg tttaacatgc ttttaccttt 120
cttatgttgt gtaactagaa ttgaaattct atctgtacat aattgattat ctcgcagcct 180
gattgagtta gtatttttta tggttctagt actgttcatg gtcatcatat ggcagtcact 240
aatgaaaaat gaccctgcat atgcatattg gtgctatgtt tttgaacttt tagcacaata 300
ctttgtaatc tgttgcatcc acttttgcag agaagtaaac atataggagt gagagggagg 360
gagagagaag gaaagcttgg tagcg 385
<210> 7
<211> 137
<212> DNA
<213>Artificial sequence 137
<400> 7
atattggtgc tatgtttttg aacttttagc acaatacttt gtaatctgtt gcatccactt 60
ttgcagagaa gtaaacatat aggagtgaga gggagggaga gagaaggaaa gcttggtagc 120
gatggggaag gagcaag 137
<210> 8
<211> 1175
<212> DNA
<213>Artificial sequence 1175
<400> 8
gcctactgct gcctgtgaat tggccacggc agggcattct tcaaaacacc gcaaaattcc 60
cttgtatttt gagtttttaa gttttaatct ttaacatgca ttttgttaca agttaaaatt 120
tattaaaaac tacttcatct gttcttattt ataagataaa tttattaata taaactaaat 180
cagttttttt ttataatatt gatagaaaca gagggagtat aaaatcatta tagttaaact 240
ttataaaatc ataaatgaaa ctcattaaac aaagagtaaa gattcacaat tttatgattt 300
ttaataaatt ttaataaata acaaaaaatt tatttaaaag cctttattca tctaaaatca 360
ttaaagtgta agttactaat atttctctta cattataaga tcttttttca agttgtcttt 420
tgcattaaat attttttaat cgaaatatct ttaattactt tacaatgaat gttataaatt 480
aaaaagataa atgcattatt tgtcttataa ggattaaaga ataagactag catataatca 540
ttaattagat aatgaataat tgagtgaaaa gaaaaacatg agtctcaata attttaaggg 600
taattttgaa aagataatag aaattgctaa aatatttaat gttactaaaa attaatcaat 660
tttcttaaaa aaatatgaat tagtgaaaag atcttatact tagagatcac gagggtatca 720
tctaattaac ttgaatattt ttttggtata taatagtgac gatgataaaa acctctgttg 780
tatttgaaaa catataaaca aaatgtctct tccaatcctt aatcagcttg caaacttcaa 840
ttatggcacg cacattagta cgttacacta tgttattagt gagttggagt gtaggaatta 900
ttattggaac acaccgaaat aaacgtaaaa atactatgta gtggtgaatg caaaaaaaaa 960
aaaaaaaatt ggcaaagaaa ttaattagag agttaataat atccaacgag caattaagga 1020
aaaaggcatc gtaggagcac aatagctgtt gaatggagcg aagagagcgc gaaaagctaa 1080
ggtatatgct tgtcgaatat tttggacaag tgtggttttc cttttcccat gttctgttat 1140
attctataaa tcattggcca ctctccattc acctc 1175
<210> 9
<211> 31
<212> DNA
<213>Artificial sequence
<400> 9
atgctctaga gggcaagaga agcaaagaga g 31
<210> 10
<211> 30
<212> DNA
<213>Artificial sequence
<400> 10
atgcccatgg cttgctcctt ccccatcgct 30
<210> 11
<211> 30
<212> DNA
<213>Artificial sequence
<400> 11
atgctctaga gcctactgct gcctgtgaat 30
<210> 12
<211> 30
<212> DNA
<213>Artificial sequence
<400> 12
atgcccatgg cttgctcctt ccccatcgct 30
<210> 13
<211> 31
<212> DNA
<213>Artificial sequence
<400> 13
atgctctaga gttgaatgga gcgaagagag c 31
<210> 14
<211> 30
<212> DNA
<213>Artificial sequence
<400> 14
atgcccatgg cttgctcctt ccccatcgct 30
<210> 15
<211> 31
<212> DNA
<213>Artificial sequence
<400> 15
atgctctaga ttgttgcatc gaatccttct a 31
<210> 16
<211> 30
<212> DNA
<213>Artificial sequence
<400> 16
atgcccatgg cttgctcctt ccccatcgct 30
<210> 17
<211> 30
<212> DNA
<213>Artificial sequence
<400> 17
atgctctaga gatgtttctt ttatatccca 30
<210> 18
<211> 30
<212> DNA
<213>Artificial sequence
<400> 18
atgcccatgg cttgctcctt ccccatcgct 30
<210> 19
<211> 31
<212> DNA
<213>Artificial sequence
<400> 19
atgctctaga atgtaactac cgtgctcgct c 31
<210> 20
<211> 30
<212> DNA
<213>Artificial sequence
<400> 20
atgcccatgg cttgctcctt ccccatcgct 30
<210> 21
<211> 35
<212> DNA
<213>Artificial sequence
<400> 21
atgctctaga atattggtgc tatgtttttg aactt 35
<210> 22
<211> 30
<212> DNA
<213>Artificial sequence
<400> 22
atgcccatgg cttgctcctt ccccatcgct 30
<210> 23
<211> 30
<212> DNA
<213>Artificial sequence
<400> 23
atgctctaga gcctactgct gcctgtgaat 30
<210> 24
<211> 30
<212> DNA
<213>Artificial sequence
<400> 24
atgcccatgg gaggtgaatg gagagtggcc 30
<210> 25
<211> 20
<212> DNA
<213>Artificial sequence
<400> 25
gaacactttc agggcaactc 20
<210> 26
<211> 21
<212> DNA
<213>Artificial sequence
<400> 26
gtcatcacag tctttgcatc g 21
<210> 27
<211> 21
<212> DNA
<213>Artificial sequence
<400> 27
tgttgatgtg caggtatcac c 21
<210> 28
<211> 20
<212> DNA
<213>Artificial sequence
<400> 28
gtcaccacgg tgatatcgtc 20
<210> 29
<211> 20
<212> DNA
<213>Artificial sequence
<400> 29
tgcaaaggag gctgctaact 20
<210> 30
<211> 20
<212> DNA
<213>Artificial sequence
<400> 30
cagcatcacc gttcttcaaa 20
Claims (3)
1. phosphate transporter geneGmPT5Expression and responding low-phosphor side of the promoter in driving legume gene in root nodule
The application in face.
2. application according to claim 1, which is characterized in that the phosphate transporter geneGmPT5Promoter sequence
Row are as shown in SEQ ID NO.1.
3. application according to claim 1, which is characterized in that the phosphate transporter geneGmPT5 containsIt is 7 different long
The promoter section of degree, each sector sequence is as shown in SEQ ID NO.2-8.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810598852.3A CN108753781A (en) | 2018-06-12 | 2018-06-12 | The application of phosphate transporter gene GmPT5 promoters |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810598852.3A CN108753781A (en) | 2018-06-12 | 2018-06-12 | The application of phosphate transporter gene GmPT5 promoters |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN108753781A true CN108753781A (en) | 2018-11-06 |
Family
ID=64020977
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810598852.3A Pending CN108753781A (en) | 2018-06-12 | 2018-06-12 | The application of phosphate transporter gene GmPT5 promoters |
Country Status (1)
| Country | Link |
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| CN (1) | CN108753781A (en) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102757969A (en) * | 2012-06-21 | 2012-10-31 | 华南农业大学 | Phosphorus transportprotein gene GmPT5 related to phosphorus transport of soybean nodulation and application thereof |
| CN106480032A (en) * | 2016-08-11 | 2017-03-08 | 中国农业科学院作物科学研究所 | The promoter of Soybean Root and root nodule specifically expressing and its application |
-
2018
- 2018-06-12 CN CN201810598852.3A patent/CN108753781A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102757969A (en) * | 2012-06-21 | 2012-10-31 | 华南农业大学 | Phosphorus transportprotein gene GmPT5 related to phosphorus transport of soybean nodulation and application thereof |
| CN106480032A (en) * | 2016-08-11 | 2017-03-08 | 中国农业科学院作物科学研究所 | The promoter of Soybean Root and root nodule specifically expressing and its application |
Non-Patent Citations (2)
| Title |
|---|
| LU QIN等人: "The High-Affinity Phosphate Transporter GmPT5 Regulates Phosphate Transport to Nodules and Nodulation in Soybean", 《PLANT PHYSIOL.》 * |
| TENG WAN等: "Transgenic approaches for improving use efficiency of nitrogen,phosphorus and potassium in crops", 《JOURNAL OF INTEGRATIVE AGRICULTURE》 * |
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