CN100350045C - Corn tyrosin protein phosphatase gene and its coding protein and use - Google Patents
Corn tyrosin protein phosphatase gene and its coding protein and use Download PDFInfo
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- CN100350045C CN100350045C CNB031539416A CN03153941A CN100350045C CN 100350045 C CN100350045 C CN 100350045C CN B031539416 A CNB031539416 A CN B031539416A CN 03153941 A CN03153941 A CN 03153941A CN 100350045 C CN100350045 C CN 100350045C
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/10—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in agriculture
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- Y02A40/132—Plants tolerant to drought
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Abstract
The present invention discloses a corn tyrosine protein phosphatase gene and the encoding protein and the application thereof, which aims to provide corn tyrosine protein phosphatase regulating and controlling the biological synthesis of ABA, and an encoding gene and the application of the corn tyrosine protein phosphatase. The corn tyrosine protein phosphatase encoding gene ZmRSPTP1 provided by the present invention is one of the following nucleotide sequences: 1) SEQ ID No. 1 in a sequence list; 2) a nucleotide sequence of a protein sequence of encoding SEQ ID No. 2; 3) a DNA sequence which has more than 90% of homology with the DNA sequence limited by SEQ ID No. 1 of the sequence list. The present invention also provides the encoding protein of the gene, particularly the corn tyrosine protein phosphatase ZmRSPTP1. The tyrosine protein phosphatase gene ZmRSPTP1 of the present invention has a wide application prospect in a plant drought resistant field, and can be used for breeding drought resistant plants, particularly drought resistant corn. The present invention has large economic benefit potential.
Description
Technical field
The present invention relates to a kind of corn tyrosine phosphatase gene and proteins encoded and application in the plant genetic engineering field.
Background technology
Drought and water shortage is one of principal element of the farming of restriction northern China wide geographic area, woods, animal husbandry production.A large amount of in recent years studies show that, plant has the ability to quick perception of drought stress and active reaction, and the acquisition of this quick perception and active reaction ability realizes by a series of adverse circumstance information exchanging processes.Plant can produce various degeneration-resistant physiology, biochemical reaction by a series of information transmission, to tide over the adverse environment of of short duration arid.(abscisic acid ABA) as one of five big class plant hormones, is acknowledged as dormin, plays crucial effects to improving stress resistance of plant, and people also generically are referred to as " abscisic acid ".ABA not only can be used as long range signals, is transported to plant leaf by root, regulation and control air vent switch, thereby the water relation of regulation and control plant integral body, and also ABA can also mediate numerous expression of gene as cell signal, to improve the resistance of plant.Why ABA can be that environment-stress such as water deficit can reach the accumulation of inducing ABA in large quantities fast as its basic foundation of adverse circumstance semiochemicals.Water deficit induces the ABA cumulative process to be actually a cell adverse circumstance information exchanging process, and this process comprises the regulation and control that cell is expressed identification, cell signalling and the ABA biosynthesizing key gene of water deficit etc.Water deficit induces the ABA accumulation to comprise the information linking of most critical in the whole cell adverse circumstance information transfer system, the research of this information linking is not only helped to disclose the identification and the transduction mechanism of the former first ringing of water deficit, and might provide direct means or strategy for the resistance of handling plant by genetically engineered.
Laboratory, inventor place induces the cell adverse circumstance signal of ABA accumulation to launch a large amount of research around water stress in recent years, but do not find signal component or signalling system that people know, can participate in the cell signal transmission that water stress is induced the ABA accumulation as Ca2+, CaM, IP3 and DG, PKC, CDPK, MAPK; Found that but water stress inductive ABA accumulation can be reduced single-minded inhibitor blocking-up (PlantCell and Environment 2000, the 23:1389-1395 and the Science Bulletin 2003,48 (4): 369-373) of agent and tyrosine phosphatase (PTP).PTK (tyrosine protein kinase)/reversible protein phosphorylation of the catalytic tyrosine of PTP all plays important effect in the signal transmission of animal and yeast cell, for example PTP can be used as acceptor and discerns many extracellular signals, and PTP can regulate the growth of cell and differentiation etc. with activated growth factor receptor precursor reactant in some born of the same parents.But people have ignored the research to the plant tyrosine phosphatase always for a long time, think and do not exist by the reversible phosphorylation of the catalytic tyrosine protein of PTK/PTP system in the vegetable cell, just had by chance in the report vegetable cell in several years up to date to have PTP, but its function is not also had deep understanding.In view of single-minded inhibitor of PTP and reductive agent drought-induced ABA accumulation capable of blocking, therefore and the reversible phosphorylation of the catalytic tyrosine protein of PTK/PTP system usually is subjected to redox regulation and control in zooblast, thinks that PTP to the DTT sensitivity may participate in the regulation and control that the ABA accumulation is induced in water deficit.
The source problem that rises of ABA signal is that the molecular mechanism problem that water deficit induces ABA to accumulate is the key problem that the ABA signal transmits, and also is the key issue that improves plant drought resistance.
The innovation and creation content
The purpose of this invention is to provide a kind of participation regulation and control biosynthetic corn tyrosine phosphatase of ABA and encoding gene thereof.
Corn tyrosine phosphatase encoding gene ZmRSPTP1 provided by the present invention is one of following nucleotide sequences:
1) the SEQ ID № in the sequence table: 1;
2) coding SEQ ID №: the nucleotide sequence of 2 protein sequences;
3) with sequence table in SEQ ID №: the dna sequence dna that 1 dna sequence dna that limits has 90% above homology.
Corn tyrosine phosphatase ZmRSPTP1 provided by the present invention is SEQ ID № in the sequence table: 2 amino acid residue sequence or with SEQ ID №: 2 amino acid residue sequence is through replacement, disappearance or the interpolation of one or several amino-acid residue and have the № with SEQ ID: 2 is identical active by SEQ ID №: 2 deutero-protein.
SEQ ID № in the sequence table: 1 by 1122 based compositions.
SEQ ID № in the sequence table: the 2nd, the protein of forming by 373 amino-acid residues.
Contain expression carrier of the present invention and clone and all belong to protection scope of the present invention.
As long as expression amount and the activity thereof of control tyrosine phosphatase ZmRSPTP1 just can be handled ABA in the intravital accumulation of plant, thereby solve the drought resistance problem of plant.Tyrosine phosphatase gene ZmRSPTP1 of the present invention has broad application prospects in the plant drought field, can be used for cultivating particularly drought-resistant corn of drought-resistant plant, and its economic efficient latent is huge.
Description of drawings
Fig. 1 is the SDS/PAGE electrophorogram of each protein ingredient in the protein purification process
Fig. 2 is the SDS-PAGE electrophorogram behind the ZmRSPTP1-His fusion rotein preliminary purification
Fig. 3 is PAO and the DTT histogram that influences to fusion rotein ZmRSPTP1-His enzymic activity
Embodiment
The extraction of embodiment 1, PTP and purifying and N-end sequence are measured
1) extraction of total protein in the maize bud scale
Corn seed (agricultural university 4967) is broadcast in the dish of 8cm * 30cm * 60cm that vermiculite and spun yarn mixture (2: 1) are housed after soaking vernalization.28 ℃ of dark culturing 5 days when coleoptile length is the 3cm left and right sides, is got coleoptile and are carried out following various experiment.
The 200g coleoptile is with 3 times of volume Buffer A (50mM MOPS pH 7.4, the bright arrestin enzyme of 2 μ g/ml peptide, 2 μ g/ml pepatatin A, 1mM PMSF, 1mM EDTA, 10% glycerine, 0.1%triton x-100) after the grinding homogenate, 4 layers of filtered through gauze, centrifugal 30 minutes of 10000xg, supernatant liquor is directly used in protein purification with Sephadex G-25 desalination.
2) with in yin, yang ion exchange column and the molecular sieve purification maize bud scale to the PTP component of DTT sensitivity
1. separate the PTP component with the yin, yang ion-exchange chromatography
(i) DEAE-Sepharose column chromatography
(3.5 * 20cm) use Buffer B balance (50mM MOPS pH 7.4 to the DEAE-Sepharose post, the bright arrestin enzyme of 2 μ g/ml peptide, 2 μ g/ml pepatatin A, 1mM PMSF, 1mM EDTA, 5% glycerine), behind the sample upper prop, being eluted to the outflow protein concentration in advance with 2000ml Buffer B is zero.1500ml 0-0.7M NaCL gradient elution, every pipe is collected 10ml, is that substrate is measured phosphatase activity with pNPP, activity is managed solution (DEAE elutriant) merge.
(ii) SP-Sepharose column chromatography
The Buffer C dialysis (50mM sodium acetate buffer, pH 4.7,1mM PMSF, 1mM EDTA, 5% glycerine) of above-mentioned DEAE elutriant.(2.6 * 20cm) use Buffer C balance, last sample to the SP-Sepharose post.It is zero that Buffer C is eluted to protein concentration in advance.0-0.6M NaCL gradient elution.Every pipe is collected 5ml, is that substrate is measured phosphatase activity with pNPP, activity is managed solution (SP elutriant) merge.Detect of the reaction of each SP eluant component phosphatase activity, will be concentrated into 5ml with 10KD ultrafiltration pipe the SP eluant component of DTT sensitivity to DTT (1mM).
2. use sieve chromatography purifying PTP component
1. the PTP component that obtains in is carried out Sephacryl S-200 sieve chromatography twice.Sephacryl S-200 (2.6 * 100cm) use Buffer C balance, the sample upper prop, and Buffer C wash-out, every pipe is collected 3ml.Is that substrate is measured phosphatase activity with pNPP, activity is managed solution (S-200 elutriant) merge that 10KD ultrafiltration pipe is concentrated into 0.5ml, cross for the second time molecular sieve Sephacryl S-200 (1 * 80cm), Buffer C wash-out, every pipe is collected 0.5ml.With pNPP is that substrate is measured phosphatase activity, activity is managed solution merge.
3. the detection of PTP component purity
After the PTP component that 2. step is obtained is carried out SDS-PAGE (12% glue) according to ordinary method with the small-sized colloid system of BIO-RAD albumen, with silver dyeing.The result as shown in Figure 1, A is a total protein, B obtains having the PTP enzyme active component through the SDS-PAGE electrophoresis result behind the DEAE-Sepharose sieve chromatography in 1.; C for the PTP component that behind the SPSepharose sieve chromatography, has the DTT sensitivity that obtains in 1. through the SDS-PAGE electrophoresis result; D be step obtain in 2. behind twice Sephacryl S200 sieve chromatography, have the PTP component of DTT sensitivity through the SDS-PAGE electrophoresis result.Silver dyeing shows that the sample in the D swimming lane is a protein band that molecular weight is 42kD, illustrates that albumen has obtained the purifying of homogeneous.
4. the PTP component being carried out enzymic activity detects
The total reaction system is 50 μ l, wherein contain 35 μ l reaction buffer (50mM MOPS, pH 6.0,1mMEDTA, 10% glycerine, 500 μ M PMSF), 5 μ l protein phosphatase enzyme substratess (2mM universal substrate pNPP or 1mM PTPase substrate END (pY) INASL and Tyrosine O-phosphate), 10 μ l protein samples, adding DTT, to make its final concentration be 1mM.Be reflected at 25 ℃ and carried out 30 minutes, add Pi analytical reagent termination reaction.The Pi that discharges is with the colour developing of Malachite-molybdate dyestuff, and 650nm measures absorption value.The protein phosphatase enzymic activity is calculated with the amount that unit time unit's protein sample discharges Pi.The protein phosphatase enzymic activity that records behind the purifying is 1200nmol pi min
-1Mg
-1Albumen.This PTP component called after ZmRSPTP1 (reducer sensitive PTP1).
3) the N-end sequence is measured
The protein sample that obtains through the above-mentioned steps purifying carries out N-terminal portions amino acid sequencing, uses the Edman edman degradation Edman to claim phenyllisothiocyanate method (being called for short the PTH method) to carry out the N-end sequence again and measures.The N-end sequence that obtains is MNCLQNLLKEPPIVGS.
The clone of the cDNA of embodiment 2, ZmRSPTP1 coding
(1) extraction of the total RNA of maize bud scale
(article No.: method 15596-026) is carried out the extraction of the total RNA of maize bud scale according to Invitrogen company total RNA extraction reagent box (TRIzol Reagent).
(2) separation and purification of maize bud scale mRNA
(PolyAtract mRNA Isolation System III (withMagnetic Stand) article No.: method Z5300) is carried out the separation and purification of maize bud scale mRNA according to Promega company test kit.
(3) reverse transcription is synthetic
According to Invitrogen Corporation's Super Script
TMRNaseH
-The Reverse Transcriptase description of product carries out reverse transcription.
(4) pcr amplification ZmRSPTP1
With reverse transcription synthetic cDNA is template, according to the N-end sequence measurement result (MNCLQNLLKEPPIVGS) of PTP, as follows from 6 amino acid of N end and 8 upstream primers of corn codon-bias design: ATGAACTGCTTGCAG; ATGAACTGCTTCCAG; ATGAACTGCCTCCAG; ATGAACTGCCTGCAG; AACTGCTTGCAGAAC; AACTGCTTCCAGAAC; AACTGCCTCCAGAAC and AACTGCCTGCAGAAC.Downstream primer is: TTTTTTTTTTTTTTTA; TTTTTTTTTTTTTTTG and TTTTTTTTTTTTTTTC.Consisting of of PCR reaction system: 5 μ l, 10 * Taq buffer, 4 μ l, 10 * dNTP, (each one of upstream and downstream primer carries out various combination for 1 μ l upstream primer, 1 μ l downstream primer, separately be PCR, totally 24 PCR reactions), 1 μ l template (about 5ng), 0.5 μ l Ex-Taq enzyme, 38.5 μ l H
2O.The PCR program is:
94℃ 3min
72℃ 5min
The PCR product reclaims purifying through QIAGEN GEL EXTRACTION KIT (#28704), serves the order-checking of Hai Shenggong order-checking portion.Sequence 1 in the nucleotide sequence of the ZmRSPTP1 that records such as the sequence table.Sequence 2 in the aminoacid sequence of ZmRSPTP1 such as the sequence table.Carry out protein B LAST in GenBank, AtPTPKIS1 in corn ZmRSPTP1 and the Arabidopis thaliana and the LePTPKIS1 in the tomato have higher homology as a result, and homology reaches 62% and 59% respectively.
(1) vivoexpression ZmRSPTP1 in yeast cell
1. pcr amplification ZmRSPTP1 encoding gene
With Upper and Lower is primer, introduces Sma I/Cla I site respectively on the ZmRSPTP1 encoding gene, and introduces 6 Histidines at 3 '-end.Primer sequence is as follows: Upper:5 '-CTGCCCGGGATGAACTGCCTCCAGAACC-3 '; Lower:5 '-TCAATCGATGTGGTGGTGGTGGTGGTGGCGCTCGGCGGC-3 '.The PCR program is:
94℃ 3min
72℃ 5min
2. the PCR product after will reclaiming is cut with Sma I/Cla I enzyme, and reclaims.
3. Yeast expression carrier p426GAL1 is cut with Sma I/Cla I enzyme, and reclaim.
4. the ZmRSPTP1 encoding gene after enzyme is cut and reclaimed respectively is connected with p426GAL1, is built into the expression vector p426GAL1-ZmRSPTP1-6His of corn ZmRSPTP1 encoding gene.
5. according to Gietz, RD. with the method (Gietz of RH.Schiestl, RD.and RH.Schiestl. (1995) Transforming Yeast with DNA. (Invited chapter) Methods in Molecular andCellular Biology.Vol 5, #5; 255-269) transformed yeast.
The result as shown in Figure 2, at the fusion rotein of yeast expression, by the ProBond of Invitrogen company
TMPurification System preliminary purification obtains the protein band (shown in Fig. 2 arrow) of 42kD, shows that the recombinant protein that contains 6 * His label not only expressed in yeast, and has obtained purifying.Among Fig. 2, A, B are the ZmRSPTP1-His fusion rotein of preliminary purification; M is a standard molecular weight.Also detect the expression of ZmRSPTP1-DHA (Double Hemagglutinin) fusion rotein in yeast cell by the routine immunization trace in addition.Wherein, detect the anti-monoclonal antibody 3f10 (#1867423) of DHA with Roche company high-affinity.
(2) activity of ZmRSPTP1 detects
According among the embodiment 1 2) method 4. the fusion rotein ZmRSPTP1-His of preliminary purification carried out the PTP enzymic activity detect.The result shows to obtain very enzymatic activity high (780nmol Pi mg as shown in Figure 3
-1Protein min
-1) tyrosine phosphatase, and enzyme work can be reduced agent 1mM DTT (dithiothreitol) and suppress at least 75%, suppressed more than at least 50% by the single-minded inhibitor PAO of PTP (phenylarsine oxide) 100 μ M.Among Fig. 3, C is contrast; PAO is 100 μ M PAO chemicals treatment; DTT is a 1mM DTT chemicals treatment.The enzyme characteristic of the ZmRSPTP1-His fusion rotein of vivoexpression is in full accord with the PTP 42kDa protein characteristic of the reductive agent sensitivity that is purified to from the maize bud scale.
Sequence table
<210>2
<211>1122
<212>DNA
＜213〉Zea corn (Zea mays L.)
<400>1
atgaactgcc?tccagaacct?gctcaaggag?cctccaatcg?tggggtccag?gtcgatgagg 60
cggccctctc?cgctgaatct?ggcgatggtc?cgcggcggca?gtcgccgatc?aaacaccgtc 120
aaaactttgc?aggcccctgg?ggcgtccact?tccggtgccg?agagcagcgc?cgtggagatg 180
ggcaccgaga?agtccgaagt?gtacagcact?aacatgacgc?aggctatggg?agcagcgttg 240
acatatagac?atgaactcgg?gatgaactac?aatttcatac?gcccagactt?gattgtagga 300
tcctgcttac?agagtccact?tgatgttgat?aagcttcgga?agattggtgt?caaaactgta 360
ttctgcttgc?agcaagattc?agatcttgaa?tattttggag?tcgacatccg?tgccattcaa 420
gattattctc?tacaattcaa?agatattgtg?cactgccgtg?cggaaattag?ggattttgat 480
gcttttgatt?tgcgattgag?gcttcctgct?gtggttagca?aattgcacaa?acttatcaac 540
tgtaatggtg?gtgtaacata?tatacattgt?actgctggac?ttggaagagc?tcctgctgtt 600
gcattggctt?atatgttctg?gattcttggg?tacagtctca?atgaaggaca?tcggctgcta 660
cagagtaaaa?gggcttgctt?tccgaagttg?gaagccatta?agttggcaac?tgctgacatt 720
ctgacaggat?tatccaaaaa?cacaatcact?ttgaagtggg?aagctgatgg?ttcttcctct 780
gttgaaattt?ctgggctcga?cattggctgg?ggtcagagaa?ttcctttgac?atatgatgag 840
gagaaaggag?cttggtttct?tgagaaagag?ttgcctgaag?gacggtatga?atacaaatac 900
gtagtggatg?gcaaatggct?atgcaacgag?catgagctga?taacgaaacc?gaatgctgac 960
ggccacgtga?acaactatgt?tcaggtctcc?agagacggca?cgagcgatga?agagaaggag 1020
cttagggagc?ggttgactgg?tcgggaccct?gatctcacgg?accaggagag?gctgatgatc 1080
agagagtact?tggaacagta?cgcggatgcc?gccgagcgct?ag 1122
<210>2
<211>373
<212>PRT
＜213〉Zea corn (Zea mays L.)
<400>2
Met?Asn?Cys?Leu?Gln?Asn?Leu?Leu?Lys?Glu?Pro?Pro?Ile?Val?Gly?Ser
1 5 10 15
Arg?Ser?Met?Arg?Arg?Pro?Ser?Pro?Leu?Asn?Leu?Ala?Met?Val?Arg?Gly
20 25 30
Gly?Ser?Arg?Arg?Ser?Asn?Thr?Val?Lys?Thr?Leu?Gln?Ala?Pro?Gly?Ala
35 40 45
Ser?Thr?Ser?Gly?Ala?Glu?Ser?Ser?Ala?Val?Glu?Met?Gly?Thr?Glu?Lys
50 55 60
Ser?Glu?Val?Tyr?Ser?Thr?Asn?Met?Thr?Gln?Ala?Met?Gly?Ala?Ala?Leu
65 70 75 80
Thr?Tyr?Arg?His?Glu?Leu?Gly?Met?Asn?Tyr?Asn?Phe?Ile?Arg?Pro?Asp
85 90 95
Leu?Ile?Val?Gly?Ser?Cys?Leu?Gln?Ser?Pro?Leu?Asp?Val?Asp?Lys?Leu
100 105 110
Arg?Lys?Ile?Gly?Val?Lys?Thr?Val?Phe?Cys?Leu?Gln?Gln?Asp?Ser?Asp
115 120 125
Leu?Glu?Tyr?Phe?Gly?Val?Asp?Ile?Arg?Ala?Ile?Gln?Asp?Tyr?Ser?Leu
130 135 140
Gln?Phe?Lys?Asp?Ile?Val?His?Cys?Arg?Ala?Glu?Ile?Arg?Asp?Phe?Asp
145 150 155 160
Ala?Phe?Asp?Leu?Arg?Leu?Arg?Leu?Pro?Ala?Val?Val?Ser?Lys?Leu?His
165 170 175
Lys?Leu?Ile?Asn?Cys?Asn?Gly?Gly?Val?Thr?Tyr?Ile?His?Cys?Thr?Ala
180 185 190
Gly?Leu?Gly?Arg?Ala?Pro?Ala?Val?Ala?Leu?Ala?Tyr?Met?Phe?Trp?Ile
195 200 205
Leu?Gly?Tyr?Ser?Leu?Asn?Glu?Gly?His?Arg?Leu?Leu?Gln?Ser?Lys?Arg
210 215 220
Ala?Cys?Phe?Pro?Lys?Leu?Glu?Ala?Ile?Lys?Leu?Ala?Thr?Ala?Asp?Ile
225 230 235 240
Leu?Thr?Gly?Leu?Ser?Lys?Asn?Thr?Ile?Thr?Leu?Lys?Trp?Glu?Ala?Asp
245 250 255
Gly?Ser?Ser?Ser?Val?Glu?Ile?Ser?Gly?Leu?Asp?Ile?Gly?Trp?Gly?Gln
260 265 270
Arg?Ile?Pro?Leu?Thr?Tyr?Asp?Glu?Glu?Lys?Gly?Ala?Trp?Phe?Leu?Glu
275 280 285
Lys?Glu?Leu?Pro?Glu?Gly?Arg?Tyr?Glu?Tyr?Lys?Tyr?Val?Val?Asp?Gly
290 295 300
Lys?Trp?Leu?Cys?Asn?Glu?His?Glu?Leu?Ile?Thr?Lys?Pro?Asn?Ala?Asp
305 310 315 320
Gly?His?Val?Asn?Asn?Tyr?Val?Gln?Val?Ser?Arg?Asp?Gly?Thr?Ser?Asp
325 330 335
Glu?Glu?Lys?Glu?Leu?Arg?Glu?Arg?Leu?Thr?Gly?Arg?Asp?Pro?Asp?Leu
340 345 350
Thr?Asp?Gln?Glu?Arg?Leu?Met?Ile?Arg?Glu?Tyr?Leu?Glu?Gln?Tyr?Ala
355 360 365
Asp?Ala?Ala?Glu?Arg
370
Claims (8)
1, from the tyrosine phosphatase encoding gene ZmRSPTP1 of corn, be one of following nucleotide sequences:
1) the SEQ ID № in the sequence table: 1;
2) coding SEQ ID №: the nucleotide sequence of 2 protein sequences.
2, gene ZmRSPTP1 according to claim 1 is characterized in that: described ZmRSPTP1 is the SEQ ID № in the sequence table: 1.
3, from the tyrosine phosphatase ZmRSPTP1 of corn, be SEQ ID № in the sequence table: 2 amino acid residue sequence or with SEQ ID №: 2 amino acid residue sequence is through replacement, disappearance or the interpolation of one or several amino-acid residue and have the № with SEQ ID: 2 is identical active by SEQ ID №: 2 deutero-protein.
4, corn tyrosine phosphatase ZmRSPTP1 according to claim 3 is characterized in that: it is the SEQ ID № in the sequence table: 2.
5, contain the described expression carrier of claim 1.
6, the clone that contains the described gene of claim 1.
7, the application of the described gene of claim 1 in cultivating drought-resistant plant.
8, application according to claim 7 is characterized in that: the application of described gene in cultivating drought-resistant corn.
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| CNB031539416A CN100350045C (en) | 2003-08-21 | 2003-08-21 | Corn tyrosin protein phosphatase gene and its coding protein and use |
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| CNB031539416A CN100350045C (en) | 2003-08-21 | 2003-08-21 | Corn tyrosin protein phosphatase gene and its coding protein and use |
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| CN100350045C true CN100350045C (en) | 2007-11-21 |
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| CN103060285B (en) * | 2011-10-21 | 2014-03-26 | 华中农业大学 | Application of OsPP18 gene on control of rice drought resistance |
| CN110643589B (en) * | 2019-09-19 | 2021-08-06 | 华中农业大学 | Protein for improving drought resistance of plants and application thereof |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2003056029A2 (en) * | 2002-01-04 | 2003-07-10 | Aventis Pharma Deutschland Gmbh | Highly sensitive and continuous protein-tyrosine-phosphatase (ptpase) test using 6,8 difluoro-4-methyl-umbelliferylphosphate |
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Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2003056029A2 (en) * | 2002-01-04 | 2003-07-10 | Aventis Pharma Deutschland Gmbh | Highly sensitive and continuous protein-tyrosine-phosphatase (ptpase) test using 6,8 difluoro-4-methyl-umbelliferylphosphate |
Non-Patent Citations (2)
| Title |
|---|
| Water stress-induced abscisic acid accumulation in relation toreducing agents and sulfhydryl modifiers in maize plant. W. Jia,J. Zhang.Plant,Cell & Environment,Vol.23. 2000 * |
| 酪氨酸蛋白磷酸酶可能影响ABA的积累和参与植物细胞水分胁迫信号传递 邢宇等.科学通报，第48卷第4期 2003 * |
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