CN108752441A - A kind of plant glutelin sorting GAP-associated protein GAP OsGPA5 and its encoding gene and application - Google Patents

Abstract

The invention discloses a kind of plant glutelin sorting GAP-associated protein GAP OsGPA5 and its encoding gene and applications.The present invention sorts mutant by Rice Glutelingpa5Phenotypic analysis and target gene Primary Location, final clone obtains glutelin sorting GAP-associated protein GAP OsGPA5, GAP-associated protein GAP amino acid sequence shown in SEQ ID NO.1 forms, or by the substitution of one or several amino acid residues and/or lacks and ors add and still there is glutelin to sort the relevant protein derived from sequence SEQ ID NO.1 amino acid sequence shown in SEQ ID NO.1.The glutelin sorting GAP-associated protein GAP of the present invention influences the assorting room of glutelin in paddy endosperm, the encoding gene of the albumen is imported in the plant that ripe glutelin content reduces, it can obtain the normal genetically modified plants of ripe glutelin content, therefore, albumen and its encoding gene of the present invention can be applied to genetic modification of plants.

Description

A kind of plant glutelin sorting GAP-associated protein GAP OsGPA5 and its encoding gene and application

Technical field

The invention belongs to genetic engineering field, it is related to a kind of plant glutelin sorting GAP-associated protein GAP OsGPA5 and its coding base Cause and application.

Background technology

Rice is the staple food grain of nearly half population in the world.The first time green revolution and being widely applied for hybrid paddy rice realize The two forward leaps of rice yield.In the recent period, China scientist has also been proposed the MOLECULE DESIGN mould of super ideal type Xian-round-grained rice F1 hybrids Type, to realize the third time leap of rice yield.However, since China's rice quality is not good enough, it is public to product far from meeting Matter especially Cooking Quality increasingly higher demands, so as to cause " domestic rice grain is bought securities with all one's capital " and the rice import after scoring consecutive bumper harvests Ever-increasing contradiction becomes increasingly conspicuous.Starch is most important nutriment in rice, and content, the Nomenclature Composition and Structure of Complexes are to rice Quality has a conclusive influence, therefore the improvement of rice eating-quality at present is often based on the improvement of starch, but protein Effect in rice eating-quality is formed also can not be ignored.Glutelin is most important storage protein in rice, is rice egg The preferred object of white quality-improving.Therefore, the molecular mechanism of the synthesis of parsing Rice Glutelin, transhipment, processing and accumulation is to rice Quality-improving it is most important.

57H mutant is the good genetic stocks of rice protein quality-improving research.6 regulation and control paddy have been cloned at present Albumen sorting key gene, and tentatively depict glutelin sorting molecular pathways, but glutelin sorting exact mechanism I Still know little about it.PX structural domains are by 100-120 Amino acid profile, for the first time in the compound somatoplasm of mankind's nadph oxidase Component p47 ^phox And p40 ^phox In be accredited.It is after PH (Pleckstrin homology), FYVE and ENTH structural domains it Afterwards, the another newcomer of fat signal pathway is added.Report there is no to participate in Rice Glutelin about containing PX domain proteins at present The research of sorting.

Invention content

The present invention sorts mutant by Rice Glutelingpa5Phenotypic analysis and target gene Primary Location, finally Clone obtains glutelin sorting GAP-associated protein GAP OsGPA5, thus a kind of glutelin sorting GAP-associated protein GAP of present invention offer and its coding Gene and application.

Glutelin provided by the invention sorts GAP-associated protein GAP（OsGPA3）, derive from Oryza rice（Oryza sativa var.Kitaaake）, it is as follows（a）Or（b）Protein：

（a）The protein that amino acid sequence forms shown in sequence in sequence table 1；

（b）By the amino acid sequence of sequence 1 by one or several amino acid residues substitution and/or lack and or add and The still derived protein with the function.

Sequence 1 in sequence table is made of 710 amino acid residues, and from amino terminal, the 67th to 179 is PX structural domains, 435-559 is CC structural domains.

In order to make（a）In OsGPA5 convenient for purifying, amino acid sequence shown in sequence 1 can be formed in by sequence table The upper label as shown in Table 1 of amino terminal or carboxyl terminal connection of protein.

The sequence of 1 label of table

Label	Residue	Sequence
			Poly-Arg	5-6（Usually 5）	RRRRR
Poly-His	2-10（Usually 6）	HHHHHH
			FLAG	8	DYKDDDDK
Strep-tag	8	WSHPQFEK
			c-myc	10	EQKLISEEDL

It is above-mentioned（b）In OsGPA5 can be artificial synthesized, also can first synthesize its encoding gene, then carry out biological expression and obtain.It is above-mentioned （b）In the encoding gene of OsGPA5 can be by the way that one or several amino will be lacked in DNA sequence dna shown in sequence in sequence table 2 The codon of sour residue, and/or the missense mutation of one or several base-pairs is carried out, and/or connected at its 5 ' end and/or 3 ' ends The coded sequence of label shown in table 1 obtains.

Encode the gene of above-mentioned storage protein sorting GAP-associated protein GAP（OsGPA5）Also belong to protection scope of the present invention.

The geneOsGPA5Can be following 1）Or 2）Or 3）Or 4）DNA molecular：

1）DNA molecular shown in sequence 2 in sequence table；

2）DNA molecular shown in SEQ ID NO.3 in sequence table；

3）Under strict conditions with 1）Or 2）The DNA sequence dna of restriction hybridizes and the DNA molecular of encoding said proteins；

4）With 1）Or 2）Or 3）The DNA sequence dna of restriction has 90% or more homology, and encodes glutelin sorting GAP-associated protein GAP DNA molecular.

Sequence 2 in sequence table is made of 2133 nucleotide.

The stringent condition can be in 0.1 × SSPE (or 0.1 × SSC), the solution of 0.1% SDS, 65^oIt is miscellaneous under C It hands over and washes film.

Recombinant expression carrier containing any description above gene also belongs to protection scope of the present invention.

The recombinant expression carrier of the gene can be contained with existing plant expression vector construction.

The plant expression vector includes double base agrobacterium vector and can be used for the carrier etc. of plant micropellet bombardment.The plant Object expression vector also may include 3 ' end untranslated regions of foreign gene, that is, include polyadenylation signals and any other participation MRNA is processed or the DNA fragmentation of gene expression.The bootable polyadenylic acid of polyadenylation signals is added to the 3 ' of mRNA precursor End, such as Agrobacterium crown gall nodule induction (Ti) plasmid gene (such as kermes synzyme Nos genes), plant gene（Such as soybean storage egg White gene）The non-translational region of 3 ' end transcriptions all has similar functions.

When using the gene constructed recombinant plant expression vector, any type can be added before its transcription initiation nucleotide Enhanced promoter or constitutive promoter, such as cauliflower mosaic virus（CAMV）The ubiquitin promoter of 35S promoter, corn （Ubiquitin）, they can be used alone or are used in combination with other plant promoters；In addition, using the gene of the present invention When building plant expression vector, enhancer, including translational enhancer or transcriptional enhancer also can be used, these enhancer regions can To be ATG initiation codon or neighboring region initiation codon etc., but must be identical as the reading frame of coded sequence, it is whole to ensure The correct translation of a sequence.The source of the translation control signal and initiation codon is extensive, can be natural, also may be used To be synthesis.Translation initiation region can come from transcription initiation region or structural gene.

For the ease of transgenic plant cells or plant are identified and screened, plant expression vector used can be carried out Processing can generate the enzyme of color change or the gene of luminophor as the coding that can be expressed in plant is added（Gus gene, Luciferase genes etc.）, resistant antibiotic marker（Gentamicin marker, kanamycins marker etc.）Or it is anti- Chemical reagent marker gene（Such as anti-herbicide gene）Deng.From the security consideration of genetically modified plants, any selectivity can be not added with Marker gene directly screens transformed plant with adverse circumstance.

The recombinant expression carrier can be in pCAMBIA1305.1 carriers (http://www.cambia.org/daisy/ Cambia/585 the gene is inserted into recombination between multiple cloning sites SmaI)（OsGPA5）Obtained recombinant plasmid.It is described heavy Organize plasmid concretely pCAMBIA1305.1-OsGPA5；The pCAMBIA1305.1-OsGPA3 is will be by OsGPA3 genomes Coded sequence is inserted into together with the promoter region of upstream 2724bp and the segment of downstream 3138bp by recombinant technique (Clontech companies, the Infusion recombination kits) obtained between pCAMBIA1305.1 multiple cloning sites SmaI.

It will containOsGPA5PCAMBIA1305.1 be named as pCAMBIA1305.1-OsGPA5。

Contain any description above gene（OsGPA5）Expression cassette, transgenic cell line and recombinant bacterium belong to the present invention Protection domain.

It is a further object to provide a kind of methods that cultivation glutelin sorts normal genetically modified plants.

The method provided by the invention cultivated glutelin and sort normal genetically modified plants, is by the channel genes paddy egg In the abnormal plant of white sorting, obtains glutelin and sort normal genetically modified plants；The abnormal plant of glutelin sorting is embryo The plant that glutelin precursor increases severely and reduced with ripe glutelin content in breast；The glutelin sorts normal transgenosis and plants Object is genetically modified plants of the physical efficiency by normal process as ripe glutelin before glutelin.Specifically, the gene passes through institute Recombinant expression carrier is stated to import in the abnormal plant of glutelin sorting；The glutelin sorts abnormal plantgpa5。

The albumen, the gene, the recombinant expression carrier, expression cassette, transgenic cell line or recombinant bacterium or described Method can be applied to rice breeding.

The carrier that foreign gene is expressed in plant can be guided using any type, the gene of encoding said proteins is led Enter plant cell, transgenic cell line and transfer-gen plant can be obtained.The expression vector for carrying the gene can be by using Ti The conventional biology methods such as plasmid, Ri plasmids, plant viral vector, directly delivered DNA, microinjection, conductance, agriculture bacillus mediated Plant cell or tissue are converted, and the plant tissue of conversion is cultivated into plant.The plant host being converted is either list Leaf plant can also be dicotyledon, such as：Tobacco, crowtoe, arabidopsis, rice, wheat, corn, cucumber, tomato, poplar Tree, turfgrass, lucerne place etc..

The glutelin sorting GAP-associated protein GAP of the present invention influences the assorting room of glutelin in paddy endosperm.By the albumen Encoding gene imports in the plant that ripe glutelin content reduces, and can obtain the normal transgenosis of ripe glutelin content and plant Object.The albumen and its encoding gene can be applied to genetic modification of plants.

Description of the drawings

Fig. 1 wild types Kitaake and mutantgpa5Exophenotype；

Fig. 2 wild types Kitaake and mutantgpa5SDS-PAGE and Western blot analyses；

Fig. 3 wild types Kitaake and mutantgpa5The coomassie brilliant blue staining of Endosperm during Its Development is observed；

Fig. 4 wild types Kitaake and mutantgpa5The immune colloid gold of Endosperm during Its Development is observed；

The map based cloning of Fig. 5 mutators；

The phenotypic analysis of Fig. 6 transgenosis complement strains；

Fig. 7 pCAMBIA1305.1 Vector maps.

Specific implementation mode

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.

The discovery of glutelin sorting GAP-associated protein GAP and its encoding gene in embodiment 1, rice

One, Rice Glutelin sorts mutantgpa5Phenotypic analysis

Japonica rice variety kitaake's⁶⁰The mutating strain series of seed silty are filtered out in Co radiomutants librarygpa5.With it is wild Type is compared,gpa5Be mainly characterized by seed silty, it is opaque（See Figure 1A）.Scanning electron microscope analysis confirmsgpa5Middle starch granules Loosely, it may be possible to the main reason for causing endosperm silty opaque（See Figure 1B）.gpa5The SDS-PAGE collection of illustrative plates of Seed Storage Protein is aobvious Show that 57 kDa precursors of glutelin increase, corresponding ripe glutelin acidic and alkaline subunit content reduce, while globulin Content also decreases（See Fig. 2A）；Western blot analyses further demonstrate above-mentioned variation（See Fig. 2 B）.

Develop mid-endosperm semithin section after coomassie brilliant blue staining it has been observed that wild type there are two albuminoids Body, the proteosome II of spherical proteosome I and irregular shape, proteosome II are slightly larger than proteosome I（See Fig. 3 A and 3C, black Triangle indicates proteosome I, red triangle mark proteosome II）.It is interesting thatgpa5In in addition to above two proteosome, Cell wall attachment also found a kind of paramural body class formation for being filled with a large amount of protein（See Fig. 3 B and 3D, red asterisk mark）. In addition,gpa5Middle proteosome II obviously becomes smaller compared to wild type（See Fig. 3 D, red triangle mark）.The above results showgpa5It is prominent Proteosome II dysplasia in variant, a large amount of protein are filled in the paramural body class formation that cell wall is formed about.

In order to parse the forming process of paramural body class formation, hair is observed using transmission electron microscope combination immune colloidal gold technique Educate the endosperm of mid-term（See Fig. 4）.Research shows that the rear golgiosome transport of glutelin is by a kind of transport for being referred to as dense vesicle Tool, which mediates, to be completed（Fig. 4 A, 4C and 4K）.As wild type, mutantgpa5Middle dense vesicle can be normal from golgiosome Budding（Fig. 4 B）.However, it is interesting thatgpa5In a large amount of dense vesicle mistakes be sorted into extracellular body space（Fig. 4 D）, gradually long-pending Tire out and forms big paramural body class formation（Fig. 4 E-4J）, therefore, causegpa5In proteosome II become smaller（Fig. 4 L-4N）.

In conclusion the above results confirmgpa5In be responsible for transport glutelin dense vesicle occur mistake sorting, in turn Cause a large amount of glutelin precursors that cannot be cut into ripe acidic and alkaline subunit into extracellular body space.

Two, target gene positions

1, the Primary Location of target gene

Utilize mutantgpa5With wide affine kind Dular（Purchased from Institute of Crop Science, Chinese Academy of Agricultural Science's germ plasm resource Library）Hybridization,gpa510 are chosen in the F2 segregating populations of/Dulargpa5Phenotype（Seed silty is opaque and glutelin precursor Increase）Recessive extremists, extract seed DNA, structure DNA mix pond.Utilize 175 couples of InDel of covering rice full-length genome Label carries out linkage analysis, will be responsible forgpa5The mutator of mutation type surface is located in No. 6 chromosome InDel labels M124 Between M85, physical distance 358.6kb.

2, the finely positioning of target gene

According to first positioning result, the molecular labeling on public collection of illustrative plates, and foundation are found between InDel marks M124 and M85 The Rice Genome Sequence information that NCBI is announced is marked in this section self-developing InDel.Utilize 198 recessive extremists pair Target gene has carried out finely positioning（Molecular labeling for finely positioning is shown in Table 2）.

Table 2 is used for the molecular labeling of finely positioning

Finally by target geneOsGPA5For finely positioning between InDel marks M161 and LK48, physical distance is 60.9 kb （Fig. 5 A）.By gene in the section is sequenced, it is found that there are single bases on the 4th exon of the 4th ORF Missing so that target protein translation terminates in advance（Fig. 5 B and 5C）.

Three, target geneOsGPA5Acquisition

Japonica rice variety kitaake leaf cDNAs are extracted, using cDNA as template, are carried out using primer primer1 and primer primer2 Amplified production is sequenced in PCR amplification, and sequencing result is as shown in SEQ ID NO.2, the protein such as SEQ ID of coding Shown in NO.1.

primer1：5'-ATGGCGACGCCGTCGTCGTTTTC-3'；

primer2：5'-CTATCTGTCCAATAACCAATTAAG-3'.

Protein shown in sequence 1 by sequence table is named as OsGPA5 albumen, is made of 710 amino acid residues.It will Coding OsGPA5 albumen unnamed gene beOsGPA5Gene, open reading frame is as shown in SEQ ID NO.2.

The application of embodiment 2, OsGPA5 albumen and its encoding gene

One, the structure of genome complementation carrier

By pCAMBIA1305.1 carriers（Bibliography：He Gao, Mingna Jin, et al.,Days to heading 7, a major quantitative locus determining photoperiod sensitivity and regional adaptation in rice. Proc Natl Acad Sci USA,2014, 111(46): 16337- 16342）'sEcoRI andNcoSmall fragment between I restriction enzyme sites replaces with double-stranded DNA shown in SEQ ID NO.3 in sequence table Molecule（IncludingOsGPA5The terminator sequence of 2.7 kb of promoter sequence and downstream of 2.6 kb of upstream region of gene）, obtain PCAMBIA1305.1-OsGPA5 genome complementation carriers（Sequence verification）, pCAMBIA1305.1 Vector maps are shown in Fig. 7.

Two, it is overexpressed the acquisition of genetically modified plants

1, the pCAMBIA1305.1-OsGPA5 over-express vectors for obtaining step 1 import Agrobacterium EHA105 bacterial strains（U.S.'s English Pretty company）, obtain recombinational agrobacterium.

2, the recombinational agrobacterium conversion japonica rice variety kitaake obtained using step 1（Wild type）, it is as follows：

（1）The recombinational agrobacterium thalline that step 1 obtains is taken, using N6 fluid nutrient mediums（Sigma companies, C1416）It is resuspended and adjusts Whole bacterium solution OD_600nmIt is 0.5.

（2）By the japonica rice variety kitaake of culture to one month（Wild type）Mature embryo embryo callus is in step（1） 30 min are infected in obtained bacterium solution, filter paper is transferred to the solid N6 culture mediums containing 10 g/L agar after blotting bacterium solution（Sigma Company, C1416）In, 24 DEG C co-culture 3 days；

（3）By step（2）The callus of culture is seeded in containing 10 g/L agar, and the solid of 100 mg/L hygromycin screens N6 solid mediums（Sigma companies, C1416）Upper culture 16 days（It screens for the first time）；

（4）By step（3）Healthy callus after culture is seeded in containing 10 g/L agar, and 100 mg/L hygromycin are consolidated Body screens N6 culture mediums（Sigma companies, C1416）Upper culture 15 days（Programmed screening）；

（5）By step（4）Healthy callus after culture is seeded in containing 10 g/L agar, and 100 mg/L hygromycin are consolidated Body screens N6 culture mediums（Sigma companies, C1416）Upper culture 15 days（Third time is screened）；

（6）By step（4）Healthy callus after culture is seeded in differential medium（PhytoTechnology Laboratories companies, M524）Upper differentiation, obtains T₀For plant.

3, the T that step 2 is obtained₀It is identified for plant, the total DNA of plant leaf to be measured is extracted, using primer Primer3 and primer primer4 carries out PCR amplification, and amplified production is sequenced, and is bimodal after deletion segment for transgenosis Positive plant（As a result as shown in Figure 6A）.

primer3：5'- AATTGCTTGCTTCTCCAGTGAC -3'；

primer4：5'- CACTCCTCCAAAGCGTTCCT-3'.

Three, phenotypic evaluation

In T0 generations, are turned into pCAMBIA1305.1-OsGPA5 plant respectively,gpa5It is planted in Chinese agriculture section with wild type Kitaake In institute's Transgenic studies field.As a result show in transgenic line T2 seeds transparent seed occur（Fig. 6 B）, SDS-PAGE Detect transparent seed（L1, L2 and L3）With wild type performance.Thus the opaque silty before transgenosis and paddy egg are demonstrated Cynanchum glaucescens body increases character and is controlled by OsGPA5 genes, i.e., the OsGPA5 genes are that glutelin sorts related gene.It will PCAMBIA1305.1-OsGPA5 rice transformationsgpa5Mutant can make its ripe glutelin content rise to normal level （Fig. 6 C）.

<110>Institute of Crop Science, Chinese Academy of Agricultural Science

<120>A kind of plant glutelin sorting GAP-associated protein GAP OsGPA5 and its encoding gene and application

<160> 3

<210> 1

<211> 710

<212>Amino acid

<400> 1

1 MATPSSFSSARKKAPSPPKHRHDGTSGLPFGMDWSPPPKRWEGRNTIWPHNPQTGWSYCV

61 MIPSWIAQTPEASATADNFLKSIVFYRIHVGIQSPEGISSSHGVLRRFSDFLKLSSDLKQ

121 EFPRKGIPPAPPKHAFSRINSSRVLLEERRNALEEWMQKLLSDIELSRSAPVAAFLELEA

181 AARSYYQDWNQRPSEVGSSAKSSADSSPRPDEHGSGVLSESSQMNSAFAHGNGPTGATGN

241 GMLGESILDQPNERASSMSNHRKKNHVFLEHGVRNGSIDTYKGVVSEEDHDSNPGHARKD

301 SAESIGSDLSSLRGSELSVPGVSSSLWDGPVDLPSGIDGHSQTEQFTGLDMQLLYDMDAQ

361 IILPADQRPKLTRLLISMDRRQVTAKTDMEDLIARLNQEVAVKEYLATKVKDLEVELEAT

421 KKKDKEILHQAVLTEREKITQLQWDKDELYRKYSEMESNLKIEQNEKTRVQSEKTTASGE

481 KEMLLEELETKRKEVESLQQHIGEFEAKSKADIKVLVKEVKSLRNSQKEMKKVLNQYHEE

541 KTELERIVNREKQRSTRARFSREKILHECRLLRERLQECTAKFVADEQDTMTIDLSSLPD

601 ALDLVTTSDNRIRLLVAEAQLLSRDDEQGSSDDGDNSDGKSSVTMSSEDAYVTDEETTKM

661 LSDLLIDNAQLRLRLNSLIRNAVNTAVKTEKEGSDGTVPKKTVLNWLLDR

<210> 1

<211> 2133

<212> DNA

<400> 2

1 atggcgacgc cgtcgtcgtt ttcgtcggcg aggaagaagg cgccgagccc gccgaagcac

61 cgccacgacg gcacctccgg gctgccgttc ggcatggact ggagcccgcc cccgaagaga

121 tgggagggaa ggaacaccat atggccacat aaccctcaaa ccggatggag ttactgtgtg

181 atgattcctt cttggatcgc tcagacacct gaagctagtg cgactgctga caacttcttg

241 aaatccatag ttttttacag gatacatgtt ggtatacaat ctccagaagg catcagctct

301 agccacggag ttcttcgaag gtttagtgac tttctaaagt tatcttctga tcttaagcag

361 gaatttccca gaaaaggaat cccccctgct ccaccgaagc atgccttttc aagaataaat

421 tcaagcaggg tgcttctaga agagagaagg aacgctttgg aggagtggat gcaaaagcta

481 ctttctgaca ttgagttgtc gagaagtgca cctgttgctg cttttcttga gctggaagct

541 gctgcacgct catattacca agattggaat cagcgccctt ctgaagtggg ttcttcagca

601 aaaagtagtg cagattcttc tccgcgtcct gatgaacatg gttcaggtgt tctctctgag

661 tccagtcaaa tgaattcagc ctttgctcat ggtaacggtc cgacaggagc aactggtaat

721 ggtatgctgg gagagtctat cttagatcag cccaatgagc gggctagtag catgtcaaat

781 cacaggaaaa agaaccatgt ctttttggaa catggtgtta ggaatggttc catagatact

841 tataagggag tggtttccga agaggatcac gattctaacc ctggccatgc caggaaggac

901 tctgctgaaa gtataggaag tgatttgagt tctttgaggg gcagtgaatt atcggttcct

961 ggagttagta gttccctttg ggatggtcct gtggatcttc caagtggaat tgatggacat

1021 agccaaacag aacaatttac aggtttagat atgcagcttt tatatgatat ggatgcacaa

1081 atcatccttc ctgctgatca aaggccgaag ttgactagac ttttgatcag catggacaga

1141 agacaggtga cagcaaaaac tgatatggag gatctcatag cacgactaaa tcaggaagtg

1201 gctgtcaaag aatatcttgc gacaaaggtc aaagatttgg aggttgagtt ggaagctacg

1261 aagaaaaaag acaaagagat tctacatcag gctgttttga ccgaaagaga gaaaattacc

1321 caacttcaat gggataagga tgagctctac aggaagtact ctgaaatgga gtcaaacctg

1381 aagattgaac aaaatgagaa aactcgtgtg cagtcagaga aaacaactgc tagcggtgaa

1441 aaagaaatgt tactggaaga gctagaaact aaacgaaaag aagttgagag tttgcagcaa

1501 catattggag aattcgaggc aaagtctaaa gcagatataa aagttcttgt taaagaggtg

1561 aaatctcttc ggaactccca aaaagagatg aagaaagtgc tgaatcagta tcatgaggag

1621 aagactgaat tagagaggat tgttaacagg gagaaacagc ggtcaacacg tgcaaggttc

1681 tcccgtgaaa aaattcttca cgaatgcaga ctgctccgtg agcgtctaca ggaatgtact

1741 gccaagtttg ttgcagatga gcaagacact atgactattg atttatcatc tttgcctgat

1801 gctttagatc ttgtgacgac atcagacaat agaatacgtt tgcttgttgc tgaggctcaa

1861 cttctgtcac gagatgatga acagggctct tctgatgatg gtgataattc tgatggtaaa

1921 tcctcagtta ccatgagcag tgaagatgca tatgttactg atgaagagac aacaaaaatg

1981 ttatctgatc tgcttattga caatgcacaa ctgaggttgc gcttgaattc tctcatccgt

2041 aatgctgtaa atactgctgt gaagacggag aaagaaggta gtgatggaac cgtgccaaaa

2101 aagacggtgc ttaattggtt attggacaga tag

<210> 1

<211> 12681

<212> DNA

<400> 3

1 cctgatatgt ctgattaggc gtttgaacgc ttgtttctca tattcttgtg agctgattga

61 ggatgtttat ttctgtgcag ggaactagct aaggatttcg ttgtatcggg gacagcatcg

121 gagtctctat atggtgcttg tgagtccatg tacaaaccaa acatggtatg catttgtttg

181 tttgtatgat cacatttagc ccgtgtaaca ttctttatgg gataacatca tgggctatag

241 caatttcgca tttctgtttg gatctgtata atatattata attctgttgc attgtactgt

301 ttagatttct accactattt tgaatgatag atgttacatg acaacatatt atttttcttg

361 acaaaggaac ctgaagaact ctttgagact atctcacaag ctctgcagtc atcagttgat

421 cgtgattgcc tcagtggttg gggaggcttt gttctgcttg tgtaagttca cttcactaat

481 tcaagattgg atgagtggct tcatttttct tctgttaaac agcttttgaa ttaaaatacg

541 ttatgcacca ttattttatc taccgttctg caataaattg gtagaagttc atcagtaatg

601 atttttttcc caatcaaata taattggagc tcaaagtcct accgcattcc gcattttagt

661 tgtttcagca cttcatttga gcatcaatac gtttctttta cataatcttc ttttgggttt

721 tcaggacacc aaccgaagta aaagagtgtg tgatcaaggg caggatggac taaatttatc

781 attgcgaaga gttcttggaa ccatttctga tgcctgctgt tcctggtctc tcagcattac

841 tatccgtgtt gtatgtggat tccgagggaa ccttgtaata tcaaagatgt tcattcaccc

901 gtagaagaaa ctgttgtttt cttacttttc tacgcttcct atgaatgaaa ggacttctat

961 ttgttgcatg agatgtgcac ttcttggcag tccttggcct tgatgatgac gattatactg

1021 atcagtaagc cggctactgg ttaattagct aagaatacgc gtatcgtggt gcaatgagct

1081 actcatactc acccttctca ggcatttgtt cgttcaccgt ttctatatct tggcaacact

1141 ttttctgaaa tggatgttag tgctttgtcg agtatggagc cgacgatgtg aagtctattc

1201 tgagtgcgag tcgttgcgac tcagatttta agttacagta ctaactacag ttcatagctt

1261 gtagtgctca gagttcaatg gtcttggctg ctttacaaaa attggtaaaa accgaggacg

1321 acttttggtc aattctatac aactaactaa aactgcttgt ttttcacaaa ttggtctcca

1381 atgatttcat ggaaatggag cgattttcgt tcagtactgt aaaccctagt attgcaaacc

1441 ctgttagtgc ttccttcgtt attttttact caaactacaa aaaataccca tgcgttgcat

1501 tgcaacgggt aagtgctatt ttaataataa taataataat attatatatg gtttgtggga

1561 actcgcttgg atccgtattt catctcaagt tagtatgcga gtttttttaa agagatttct

1621 tatacgactc cttttgtatt ttcaaaatca aacgagctta aaatccaact caaacacaac

1681 gaacttaaaa ccgactcaaa acgtaccaaa atttttgcaa aaacaactct aatttttata

1741 atagtagaga agagatagag atatacatgt tttttttgtt attgtttcga gcatcaatac

1801 aatctcaaat atgtgtttgg tcattgtttc ctccaactat agatgtattg aaccccataa

1861 ttttttctac ttagatattg tttagattct aacttttttc ttcaaacttc gtgtgtgttt

1921 agttcacgtt aaaattaaaa gtttggttga aattggaaca atgtgacgga aaagttggaa

1981 gtttatatgt gtagaaaaaa ttagaagttt atatgtgtag aaaaaattga tgtgatggaa

2041 aagttgaaag tttgaagaaa aaatttggaa ctaaactcgg ccttctaact tttccgtcac

2101 attgaacttt cgtacacaca taaacttcta actcttccgt cacatcgttt caatttcttt

2161 gaactttcaa ttttagtgtg gaactaaaca cagcctagta gtatgtactg aaccagttcg

2221 tcactaagaa gtgcatacca ttatctgtcg gcagccatgg ccacatccaa caattacact

2281 tgtgtgggct tgcgaaaaac ccacacaact ccccgccggc ccaccaccaa ctccaccacc

2341 ccacacggcg gtggaaaagc ctagacccgg accggaccgg accgacgccc tcgcttggct

2401 gccaccgccg cgcgttgacg gttagacgcc accgcgggag gagagcccct ccctccccgt

2461 ccgcgtccgc gtccgcctca tctccgccgc cctcttactc cccctcccct tcctccactc

2521 cgtccacctc ggaatcgtcg gcgcgacgcc tccaaacccc cgcatcgatt cctccctccc

2581 acctcacccc acccttccgc ccgctcgatc caccgccgaa gcgggcggcg atggcgacgc

2641 cgtcgtcgtt ttcgtcggcg aggaagaagg cgccgagccc gccgaagcac cgccacgacg

2701 gcacctccgg gctgccgttc ggcatggact ggagcccgcc cccgaagaga tgggtaagcg

2761 cgatcccccc cccccctccc ccccgctaat ttcgatccgg gctcgattga tcgggggttc

2821 ggttgggttg gctccctgct ttgatctgtg agggacgtgc ggttcgggag ttggagttgg

2881 gagcttgcga tcctgtggcg ttgctgctgc gtgcgtcaga ttcggcgtta gttgcgaggt

2941 gtcggggatt ttctttcttt ctttctttct tttaatgctt gtcctctcga tcgtaacgat

3001 atcgcgagct ggatttgccc gcggctatgt ggtgtgtaat agtagtggaa tgtaaggcta

3061 ggaattcatg tgtgggttgg gatctgcatt gggatttgtt ggtttggagt gcctgcgttg

3121 ctactgattc agagttgttt tagagctttc ccctggatag ggttttgaat tcctgtttct

3181 tttgcaagga gtcgagtatg ggtttcaggt aggactactt ctgcggggat gggtttagac

3241 aaaccgagtg cagttcagct tcatgatatg cggattgatt tggttgtata tgtgctgctt

3301 atttcaaaga ctctggtttt gtctgagtcc atcggacata agggctttgg atttctatat

3361 taggcaatta ggcatgttca tactgttacg cataaggcca taagtacaat aattgatttc

3421 gtatacatat ttgagcaggc tgatggtgat tttattattc atcattgttc ccattgtgaa

3481 gattagctct tttgtttgac ctgcctgcct tgtgctagcc tcattctttg cacttggtgt

3541 gtttcccaac atctttacca catttaacat gaaatgatcc tagaacgcaa atgaagatta

3601 acctactagt cctagtcaat tctttttcac cgtcaagtaa atcacatgcc tcgttaacaa

3661 aattcagtaa ttttcaaaca actgggaaag ttccttgaag atgtttcctt gtattaaaga

3721 aaatgtactt ccatgaagac cacataactt ctcaatctag gtttcgctgg aaagatgatg

3781 ccatatcaat aggataccat gaaaagttcc attgcttgat gcttcttacg gcttatgctt

3841 attgatctgc aggagggaag gaacaccata tggccacata accctcaaac cggatggagt

3901 tactgtgtga tgattccttc ttggatcgct cagacacctg aagctagtgc gactgctgac

3961 aacttcttga aatccatagt tgtgagtact gcttatatta tcatcacgtt atttttgtca

4021 gtcaatttgg attgacgtac agattggaaa tgttgtgtct ctggtattac cgctatgcta

4081 caacataaat gtgcttggag tggattacaa gtgagaaaac cttgttagac caataaaaag

4141 gacaaaagaa gttcagcact tatcaacgaa gaactgagct tctttcaaaa ctatctcata

4201 ttgggtagtg gatgccaact ttctaatgta ttaccttttt attgttcttt cattttgagt

4261 aaatcgtgtc actttccaat attccaccat acttatcaaa ctagtttatt aggctgctgg

4321 actcacaaaa ctgtactttt ctttagatca ctaaagggtc acttttggtt gttagcacac

4381 tgtcctgact cttaatgaca acatggcgtg ttgaggcata cccctgtatg gggtgcaaat

4441 tttgttgtga taaccaatcg ctgacgtgca gtacactaga ggtgattctc cctctccgca

4501 caactgcctc acttgtctca gtacacaggg ggtttgagtt gggggtgaga gagaactatg

4561 taaggccaaa actcatgggg gacaagtgca aatcacaata gccatggctg aagtcattaa

4621 ctcacgtgca gcccatgcca caatggtgta tgccaacatg acatgccata ggaagtattg

4681 tgctaatgat tcagggtgac ctactggcag aaatatactt tttggtgttt tggtaactgt

4741 ggtgcatctt acctctcttt tggctcacta gtttagtcgt gatcgacggg caagagacac

4801 ttatgtaatt ctgttgcata tatcatgcaa ttcctgtttg ttgtgaactt tgatactgtt

4861 ctgtttatga tctcatttta atgcccataa aggacagttt tacaggatac atgttggtat

4921 acaatctcca gaaggcatca gctctagcca cggagttctt cgaaggttta gtgactttct

4981 aaagttatct tctgatgtaa gttgagacac ctctttgcta ttttcttctt tcacatgcca

5041 agtgtcttta aattgtaccg accttctttc ctgagctaga agcccactaa cattttcagt

5101 tggaacatac ctttcaagaa caacctccag tgatttttaa ataagaataa tcccctatcg

5161 tatcctgagt ggtctgccag attaatttat ctggcattaa atttgaaaat gtaatcaaac

5221 ctttgaattg gacttacatt tgctagtgca ttgaatgcac aattgcttgc ttctccagtg

5281 acttccgttg caacttagga tttttggtgc attgcatcac acaggtagtc agacttcaga

5341 tactttaaaa catactgtta ttataaatct tgacttagtg tatacggtat gcttacctgg

5401 gccaaattag tttttacaga tttttgcatc ttccgctggt gacttcataa tgaggacttc

5461 accttttgca attttatggt ttctataatg aatgtggaca ttttggaagt ctcatcttgg

5521 atatgattct gagaaaaaaa gattattact ttccattcga cctctaatcc aatatcattt

5581 cttcattcta agattctccc gtactgcagc ttaagcagga atttcccaga aaaggaatcc

5641 cccctgctcc accgaagcat gccttttcaa gaataaattc aagcagggtg cttctagaag

5701 aggttagccg ttatgtatat tgattaattt ttgctttgtt cgatgtcaaa tactacattc

5761 agttgtgctt agcctttttg accggtggtt cttgaaatgt tttgtttgca gagaaggaac

5821 gctttggagg agtggatgca aaagctactt tctgacattg agttgtcgag aagtgcacct

5881 gttgctgctt ttcttgagct ggaagctgct gcacgctcat gtatgttccg cttgacatat

5941 tctattgaag acactttttc ctgattgtgc attgatccat ttattaacta aaatttctta

6001 acatttttag caaatgttta attacgctat ttttctgaat gaaagattac caagattgga

6061 atcagcgccc ttctgaagtg ggttcttcag caaaaagtag tgcagattct tctccgcgtc

6121 ctgatgaaca tggttcaggt gttctctctg agtccagtca aatgaattca gcctttgctc

6181 atggtaacgg tccgacagga gcaactggta atggtatgct gggagagtct atcttagatc

6241 agcccaatga gcgggctagt agcatgtcaa atcacaggaa aaagaaccat gtctttttgg

6301 aacatggtgt taggaatggt tccatagata cttataaggg agtggtttcc gaagaggatc

6361 acgattctaa ccctggccat gccaggaagg actctgctga aagtatagga agtgatttga

6421 gttctttgag gggcagtgaa ttatcggttc ctggagttag tagttccctt tgggatggtc

6481 ctgtggatct tccaagtgga attgatggac atagccaaac agaacaattt acaggtttag

6541 atatgcagct tttatatgat atggatgcac aaatcatcct tcctgctgat caaaggccga

6601 agttgactag acttttgatc agcatggaca gaagacaggt gacagcaaaa actgatatgg

6661 aggatctcat agcacgacta aatcaggaag tggctgtcaa agaatatctt gcgacaaagg

6721 ttacgttttc tacctgacat tcatttgtga tgcctatgga ttatgcttaa acacatgata

6781 tgctttgttt tatatgtcac gagtttaaaa taaattgatt tgtgtttttt tgttcatact

6841 ttattgatgt acgttgttct tgtgcttctc tgctattctg tgcaggtcaa agatttggag

6901 gttgagttgg aagctacgaa gaaaaaagac aaagagattc tacatcaggc tgttttgacc

6961 gaaagagaga aaattaccca acttcaatgg gataaggatg agctctacag gaagtactct

7021 gaaatggagt caaacctgaa gattgaacaa gttggttttc ctctttgtct cctctttgtc

7081 atcatggctt ggtttcttac tatccatctt acaagtattt tcttacattt ctttctactt

7141 aattgcagaa tgagaaaact cgtgtgcagt cagagaaaac aactgctagc ggtgaaaaag

7201 aaatgttact ggaagagcta gaaactaaac gaaaagaagt tgagagtttg cagcaacata

7261 ttggagaatt cgaggcaaag tctaaagcag atataaaagt tcttgttaaa gaggtgaaat

7321 ctcttcggaa ctcccaaaaa gagatgaaga aagtgctgaa tcagtatcat gaggagaaga

7381 ctgaattaga ggtataaatg taaatttaaa gcttttcttt ctcgcagaac tatgatagta

7441 tctataaatt gtgtgaaatt gttgctatgg ctggcctctc attgctcaaa gccaggagag

7501 cataaccctt tacaagttgc aaactagatt ttggttgtag ctaagagaaa gtggcttgtc

7561 aaagtcatta gacaagtccc cacaaaaaaa aaagtcatta gacaagagta agcctttgaa

7621 agcatgcaag ccatcttgag tgtttgcttt tccacttcta acttattacc ctgatttctg

7681 catctccacc acccatgcaa catgttgtcc atttttggaa tattatcatg aattgttgta

7741 cttgcctact tggtatctga caagatgagc ttaacagtct tcttttgatc aacaaacata

7801 tggacctttt ccctttttta ttatataacc agcacagcca tctattcttt atgtgattcc

7861 ttttgtcaag gccggctgaa taaagatttc tatctacaca agctagattc aacagcgatt

7921 tctttatcta tatataatca ctcttggagg acaattgaat taatttaatg ctacctgttt

7981 gtacaccact taactaaaga aattgagaga gagaatgaag gggtgaggtg ttcttttgtt

8041 gttttgcttt gcacactaat gatatggttg atcataagac caaggatagt gcgatcttct

8101 caggtcgtgt gggcttgtcc tggcacggct agaggctttg attagctagg gtttgggcag

8161 aaaccttacc ttttataggg ataactcaag actgctgcta tacacacccg tgggtatttt

8221 tcatgtaaat agtagtttga ccctcaagaa ggtcattact cttccgttca ggaatataag

8281 gtgtattttg actgtccaat tttgaagagc tttgacgaac tattaatcca ataatattta

8341 ggttttacca taagaaaagg atattattgg attcatgttt gaaattactt tcttatgact

8401 ctgattttga cagtgtaaaa gtaccttaaa aaacatcttt aattttccag ctaaatcaga

8461 tccttcctaa gtcctaactg atagataggg tacaaaataa ccaatcccat gctttgaaac

8521 taataattat ttaggcaata gatattcttg gaaggtatgt aaacccttcc catattctgt

8581 tctacaatat gcattactgt ccgtgcttca agcatcaacg ctcataacag ctgatttatc

8641 aacaactttg atccaatcta tccctattac cattttttgt tttcttaaat tcaagcttct

8701 ggtagttata cttcttgcaa tcggcattct ctctagctac taattgtatt tattatatat

8761 tacagaggat tgttaacagg gagaaacagc ggtcaacacg tgcaaggttc tcccgtgaaa

8821 aaattcttca cgaatgcaga ctgctccgtg agcgtctaca ggaatgtact gccaagtttg

8881 ttgcagatga gcaagacact atgactattg atttatcatc tttgcctgat gctttagatc

8941 ttgtgacgac atcagacaat agaatacgtt tgcttgttgc tgaggtaatg ttttaaatat

9001 cttttgatta gttattgtca ttcaaactcc ttctagcttc tcacgggaaa ggcacactta

9061 gaaagttgag ctcaaactcc gacggaaaca tcagacacta cctgacacta gattcttcaa

9121 aggatctgga gtcagatttg gaacaactcc tggcctaaat tgatttgtag tttcctcagc

9181 ctctgcttat gtttgaagtt tgagccacct agttgggcct tcaaagctgc attcttgatt

9241 ctattggaac aaggtttagg tgttagtttt ggttagctct ggggaacagt tttcatcttc

9301 ccagctgaag agtgatatca tttcagagtc agtaaaacta tatttaacaa aatgagtaaa

9361 atgatctata ttttcacaaa ctttttttcc tattttacag tcatagatgc atttacttta

9421 gattacatac tggcgccact atacctaaaa aagaagtctt gccaatgacc aatgtaatgg

9481 tcatcttggt acagaagaca ttgttttcag tgttatgtac ctgtccattt agaaattgat

9541 tttgaatatt caaagttacg cttctgtttt catgaagttc tcacttcctt gtgcccatga

9601 aagcggatct gttagttaca tttacattca caatctgtgt tctataatga tcctctctca

9661 atctctcagg ctcaacttct gtcacgagat gatgaacagg gctcttctga tgatggtgat

9721 aattctgatg gtaaatcctc agttaccatg agcagtgaag atgcatatgt tactgatgaa

9781 gagacaacaa aaatgttatc tgatctgctt attgacaatg cacaactgag gttgcgcttg

9841 aattctctca tccgtaatgc tgtaaatact gctgtgaaga cggagaaaga aggtagtgat

9901 ggaaccgtgc caaaaaagac ggtgcttaat tggttattgg acagatagaa aaaccagtat

9961 atattcttgt gaattagctc ttctagttat gcggccttct caaagttata tgatgagtat

10021 tgttataatg tgggagtaac aggagtggag aaaaaaaaat caaaatcttc cttacatgca

10081 gatcaagcat gtggtggtgt tgtatctctt atttacaaag attgatcaag gataatatat

10141 ttaggaggct tctgagcctg gcctcctagc gggagaacgg tagggctcat atggttttgg

10201 ggaggcttgc aactggagtg gtagagaaca tgggtccccc ttttttgcac catacttcaa

10261 tttgtaattg ccgagctgat ggatagctgc atggttgtat ctttgtacag atgtttcaga

10321 tggtgagttc ccagcaggaa agcattgtac actatagtta aactctggat taatcttctt

10381 cacccataga gtgctcaact gttaaatcag tacttcttac tcatgcagtt gcacctcttc

10441 cagactgaaa acttctcaaa gttctcatgt tgcatacagc tggtaataag agcgtaattt

10501 actcaggcat ggacacctga ccctgcctga tgtttagatc attacgtgcg tacctttata

10561 cagatggtag ccgccacctg tgtaatattt tccatgtcac tttcagtctt tcgttagagt

10621 tttcaagtcg ctttctattt tactttatta aatgttcttg acaagattaa aaggcgtatc

10681 ctgaaatttt ttatccagat gttctttttt agataataga atctctacta ttataaaaat

10741 taaaaatgtt tttgccggta ctttggtacg tcatccatgt gatcgttcgc ttttagaaat

10801 atagataagt atttgactcg gattttaagt tcgtttgttt taggaaatat aaaactaagt

10861 cgtataaaaa aatcttttaa aaaaactcac atactaactt gagataaaag ttggactcct

10921 gactgcatct catgatttgt tgcaacgcac cggtattttt ttttctagta aaagaaaaat

10981 acggatgtac tccgtagtat gggtttgttc ggtagcccaa attttacagc cgtcttgctg

11041 cattggacaa ccaattcgtt cggctgcaca caaactattg ctccctacac ttgctagctt

11101 caacatctat aggttgtagc tactattggc tagctgcagc cgctggttgt ttgcgctgta

11161 gtaggtcttc cgagttccga ataagagcat ctccaacggt ctctctaaat ttgactctct

11221 aaatgtctat ttagccaact ctccatttaa tttagcaaat caaactgtat tttactccaa

11281 cagcctctct attcatcctc tctattctga gtctatgata attgggccca catgtcagct

11341 tcgacacctc cttctccctc atttttctcc ccttctctcc aaaccccact tccctctccc

11401 atccgccgcc gccgccccat ccccatcccc accgccgctc gggctgcgtc gccatccgcc

11461 gccgctgctc gggctccgcc cgccggtgcc gctctcttcg ccggcggtgg tgcggcgacg

11521 cgaggaggcg ggcggccgcg cgcgatggtg tggcgatgcg gggaggcagt aggcaagacg

11581 ccggcggctc gcgacgacga cctcgatgtc gctcggggtg gagctcggcg gcggcggctc

11641 gcgacgacct cgacggcgga tctagagggg ggcagcagcg gagctcaggg cggagctcgg

11701 cggtggcggt ggcggtgctg gccgcgcgac gacggggcgg cggtggccag ccgtggcgac

11761 gacggggcgg cggggctggc cgcggcgacg acggggaggc gggggccggc cgcgcgacga

11821 cggggcgacg gtggccggcc gcggcgacga cgaggtgacg acgccctcgt cacatgtgca

11881 tgccgcactg ctgcatggag agagagagag ggggaaacag gcgtgggtcc cacatatggc

11941 tagccggttt tggctggcca actttagcta gtctggtggg gaatttagac agccggatgg

12001 cttggagagg catttggcta gtctgttgga gtagattttt tttgtgtaag ttagccaaat

12061 tttaacttag agagccattt ggctaggctc ttggagatgc tctaaggcct ataactacaa

12121 gatcattcag aacgatttga tctccattgt ttgcacttaa gtttaccagt acgtcgttag

12181 aaattgaatt tttaagttgc tcaaagtata tgtatataaa atttacacat tattttttcg

12241 ctattaagcg ttgctgctta tccaaacatt agcattgttt agataggtgt tgtattcgct

12301 cggtcttatc tctattctag gtttcggcaa taaacctact aaatatactt taattaaacg

12361 tatagagaaa acacctctag aactatgtat ttacggttaa agacttttgt cctacttgct

12421 agcgactcaa ttacatttat aaccttccta gccactttcc tcattttccc ctttttcaac

12481 atttcctctt cctctcccct gagggtgcgc caccctaacc cgatcttccc tatctccagc

12541 gtcacctcca tctccctacc ctacgatgcc actcaagcta tcctgttaac atagaaccat

12601 ctcgaaattg cttcgtagtc atcacgcgac gtgtgattcg ttagaatcgc tctttgcaga

12661 tctacctctc agtgtccaaa t

Claims (10)

1. a kind of glutelin sorts GAP-associated protein GAP, it is characterised in that selected from such as（a）Or（b）Shown in it is any：

（a）The protein that amino acid sequence forms shown in SEQ ID NO.1；

（b）The substitution by one or several amino acid residues and/or missing by amino acid sequence shown in SEQ ID NO.1 And/or it adds and still there is glutelin to sort the relevant protein derived from sequence 1.

2. glutelin as described in claim 1 sorts GAP-associated protein GAP, it is characterised in that its end, which increases, sequence label, preferably For Poly-Arg, Poly-His, FLAG, Strep-tag II or c-myc.

3. encoding the gene of glutelin sorting GAP-associated protein GAP described in claims 1 or 2.

4. gene according to claim 2, it is characterised in that：The gene is following 1）To 4）DNA shown in any points Son：

1）DNA molecular shown in SEQ ID NO.2；

2）DNA molecular shown in SEQ ID NO.3；

3）Under strict conditions with 1）Or 2）The DNA sequence dna of restriction hybridizes and encodes DNA points of albumen described in SEQ ID NO.1 Son；

4）With 1）Or 2）Or 3）The DNA sequence dna of restriction has 90% or more homology, and encodes glutelin sorting GAP-associated protein GAP DNA molecular.

5. the recombinant expression carrier, expression cassette, transgenic cell line containing the gene of claim 3 or 4 or recombinant bacterium.

6. recombinant expression carrier according to claim 5, it is characterised in that：The recombinant expression carrier be It is inserted into what gene described in Claims 2 or 3 obtained between multiple cloning sites EcoR I and the Nco I of pCAMBIA1305.1 carriers Recombinant plasmid.

7. albumen described in claim 1, gene described in Claims 2 or 3, recombinant expression carrier, expression described in claim 4 The application of at least one of box, transgenic cell line or recombinant bacterium in plant breeding.

8. application according to claim 6, it is characterised in that it is in cultivating glutelin and sorting normal genetically modified plants Application.

9. a kind of method cultivated glutelin and sort normal genetically modified plants, is by channel genes paddy described in Claims 2 or 3 In the abnormal plant of albumen sorting, obtains glutelin and sort normal genetically modified plants；The abnormal wherein described glutelin sorting is paddy Amyloid protein precursor abnormal accumulation.

10. according to the method described in claim 8, it is characterized in that：The gene imports glutelin by recombinant expression carrier It sorts in abnormal plant, wherein the plant is monocotyledon or dicotyledon, preferably tobacco, crowtoe, quasi- south Mustard, rice, wheat, corn, cucumber, tomato, willow, turfgrass or lucerne place.