CN1537943A - Alocasia odora agglutinin protein gene and its application - Google Patents
Alocasia odora agglutinin protein gene and its application Download PDFInfo
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Abstract
A nucleotide (cDNA) sequence of Alocasia agglutinin protein, its coding amino acid (AA) polypeptide sequence, the process for using the gene to transform cotton or other plants, and its application in medicine or other fields are disclosed. Said gene is a member of mannose conjugated agglutinin gene family and is a constitutively expressed gene.
Description
Technical field
The present invention relates to the clone and the application thereof of phytohemagglutinin protein gene, particularly relate to the cDNA sequence and the encoded protein matter sequence thereof of Alocasia ordora agglutinant protein gene.The invention further relates to of the application of Alocasia ordora agglutinin gene at engineering of insect-resistant plant.
Background technology
Phytohemagglutinin is meant the reversible vegetable-protein that is attached to special monose or oligosaccharides of also energy that contains at least one on-catalytic structural domain.
Phytohemagglutinin does not have the common structural performance.Most lectins contain covalently bound glycan molecule, belong to glycoprotein.The composition of carbohydrate is similar to other glycoprotein in the lectin molecule, mainly is seminose, N-acetylglucosamine, semi-lactosi, Fucose, sialic acid, and a small amount of lectin contains wood sugar, pectinose.Some lectin does not contain covalently bound sugar.The protein portion of most lectin molecules contains higher aspartic acid, Serine and Threonine, accounts for 30% of total amino acid content, and sulfur-containing amino acid is few or even do not have fully.The lectin molecular weight differs greatly, from several ten thousand to hundreds thousand of.A lot of aggegations have assembles tendency, is subjected to the influence of pH value of solution, temperature etc.
Phytohemagglutinin be a class tool high degree of specificity carbohydrate in conjunction with active albumen, this is the sign that they distinguish all other vegetable-proteins.Its sugared binding specificity has following characteristics: 1, and the scope of the carbohydrate that lectin can specific combination is very large.2, the lectin of different structure may be discerned identical carbohydrate.The sugared binding specificity of phytohemagglutinin is mainly determined by the three-dimensional structure of binding site.3, most of lectins have higher avidity to oligosaccharides.4, the specificity of most lectins is not the glycan molecule at vegetable cell usually, and usually is the internal surface of microorganism or insect or the glycan of outside surface.5, have stability preferably.The lectin of those involved in plant resistance mechanisms particularly.Their great majority can not degraded by the proteolytic enzyme of animal or insect stomach and intestine, and stable in very big pH scope, thermostability is strong.
The physiological function of phytohemagglutinin mainly contains: 1. defence pathogenic agent invasion, the 2. effect in symbiotic nitrogen fixation, the 3. effect in cell adhesion, 4. mediate pinosome, the 5. effect in pathogenic bacterial infection host macrophage phagocytic, 6. in seed maturity and germination process, play a role, 7. insect-resistance.
Phytohemagglutinin is widely distributed, have 14% to contain phytohemagglutinin in the higher plant approximately, ratio is up to 43% in Papilionaceae, ratio reaches 12% in the Gramineae, from plants such as pulse family, Solanaceae, Euphorbiaceae, Gramineae, Liliaceae, Amaryllidaceae, find 1000 various plants lectins altogether at present, wherein the leguminous plants lectin reaches kind more than 600, and its intraseminal phytohemagglutinin content is the highest, reaches about 10% of seed protein total content.
From evolving and the structural dependence classification, phytohemagglutinin roughly can be divided into 7 different families:
1. legume lectin element; 2. the monocotyledons mannose binding lectin mainly is present in Amaryllidaceae, Araeceae, the orchid family, Liliaceae, these five sections of Bromelia family; 3. the chitin that comprises the Hevin structural domain is conjugated protein; 4.2 type ribosome inactivating protein; 5. Curcurbitaceae phloem lectin; The plain family of 6 jackfruits; 7 Amaranthaceae lectins.
The Alocasia ordora lectin shows that through the preliminary experiment result all kinds of aphids, small white, cabbage caterpillar etc. are had certain stomach poison function, and is wherein relatively good to the bean aphid effect.Also inhibited and influence Reproduction in addition to growing of bean aphid, and suppress the bean aphid honeydew and secrete behavior.Its mechanism of action may be: the Alocasia ordora lectin by insect's food-taking after, combine with Digestive system proteolytic enzyme etc. in digestive tube that (protein contains sugar chain mostly, so lectin can combine with it), the enzyme activity that suppresses the latter, make its decomposing protein effectively, the insect source of nutrition is obstructed, finally causes insect death.
The Progress on Molecular Biology that plant genetic engineering utilization is up-to-date, with advanced genetic engineering technique with relevant gene, as the gene of pest-resistant, disease-resistant, drought resisting, salt tolerant alkali etc., in the plant materials of transduceing, and make it to be expressed.Successfully GNA GNA (Galanthus Nivalisagglutinin) gene transfer has been obtained the significant social economic benefit on tobacco, potato, paddy rice, cotton and tomato at present.
About the clone of phytohemagglutinin gene,, can search the gene order of 388 relevant phytohemagglutinins by the gene database among the NCBI (Genbank).
Only clone its protease inhibitor gene in the Alocasia ordora, before the present patent application, also do not have any report of Alocasia ordora agglutinant protein gene clone.
The present invention seeks to from Alocasia ordora, to clone the agglutinant protein gene, for the Alocasia ordora lectin applies on the transgenic pest-resistant crop, batch production production, and provide molecular basis and condition in medical research and the relevant utilization.
Summary of the invention
Alocasia ordora agglutinant protein gene provided by the invention obtains with the separation of RACE PCR method from Alocasia ordora tissue (plant), it is characterized by:
(1) has Nucleotide (cDNA) sequence of sequence 1 in the sequence table, wherein the 1st～63 bit base is for containing the sequence in promoter regulation element and the non-lectin translation of 5 ' end district, the 64th～873 bit base is opening code-reading frame (ORF), and the 874th～1124 bit base is the sequence in the non-lectin translation of 3 ' end district.
(2) the amino acid polypeptide sequence shown in the sequence 2 in the code sequence tabulation, this sequence is made up of 270 amino-acid residues, the 3 d function structural domain of two seminose specific combination before and after having, a structural domain is by Gln (glutamine Q), Asp (aspartic acid D), Asn (l-asparagine N), Val (Xie Ansuan V), Tyr (tyrosine Y) five amino acid constitutes.The position that wherein constitutes five AA of previous structural domain is respectively: Gln51, Asp53, Asn55, Val57, Tyr59, back one is: Gln170, Asp172, Asn174, Val176, Tyr178.The biological characteristics of Alocasia ordora lectin mainly shows as: specific combination seminose reversibly causes final death thereby make insect produce the apocleisis reaction.
(3) has the nucleotide sequence (cDNA sequence) of sequence 1 in the series of tables; Or with the 64th～873 bit base sequence of sequence 1 in continuous 100 zones more than the base homology more than 95% is arranged, have the nucleotide sequence of the equal functional protein of coding and sequence 2.
(4) the amino acid polypeptide sequence of coding Alocasia ordora lectin precursor protein, this polypeptide has the sequence shown in the sequence 2 in the sequence table; Or coding have same function by sequence 2 polypeptides derived sequences, or its conservative property variation polypeptide, or its active fragments.
According to Alocasia ordora agglutinant protein gene order information provided by the invention, sequence 1, sequence 2, those skilled in the art can obtain the gene that is equal to Alocasia ordora agglutinant protein AML by the following method easily.
1. with the AML gene fragment base group library acquisition of probe screening Alocasia ordora;
2. according to AML gene order information design oligonucleotides primer (containing degenerated primers), obtain with the genome of pcr amplification method from Alocasia ordora;
3. on the basis of AML gene order, obtain with the gene engineering method transformation;
4. the method with chemosynthesis obtains.
Term AML of the present invention etc. are homogenic, be to be defined as with the nucleotide sequence or the aminoacid sequence of AML gene to compare the difference that one or several Nucleotide or amino-acid residue are arranged, comprise change, shortage or the insertion of base or amino-acid residue, or with continuous 100 zones more than the base in the AML gene order homology more than 95% is arranged, and the function of its gene expression product is identical with the function of AML expression product.
Therefore, AML gene provided by the invention comprises that also AML etc. is homogenic, and they are one of following nucleotide sequences, or the nucleotide sequence of one of the following amino acid polypeptide sequence of encoding:
(1) with replacement, shortage or the interpolation of the nucleotide sequence of sequence 1 through one or several Nucleotide, tool coding and the proteinic derivatized nucleotide sequence of sequence 2 identical functions;
(2) with the 64th～873 bit base sequence of sequence 1 in the above zone of continuous 100 bases have homology more than 95%, and the nucleotide sequence that is equal to of coding and sequence 2;
(3) with the replacement of the amino acid polypeptide sequence of sequence 2 through one or several amino-acid residue, lack or add, and have derivative amino peptide sequence with sequence 2 same functions, or its conservative property variation polypeptide, or its active fragments.
AML gene provided by the invention has important use and is worth.One of application is that described AML is made up plant conversion carrier, with any method for transformation the AML gene is imported cotton or other vegetable cell, can obtain to show the genetically modified crops of the above gene, thereby reach pest-resistant effect.
The Another application of AML gene provided by the invention is that expression vector is arrived in this gene transformation, and makes it to express the generation corresponding proteins, afterwards expressing protein is applied to agricultural, medical science or others.
The present invention can produce huge economy, social effect.Cloned genes of the present invention is transformed in the plant, then can makes and do the tangible pest-resistant effect of deposits yields, particularly to the piercing-sucking mouthparts class pest.Thereby reduce using of agricultural chemicals, reduce agricultural chemicals environment, ecology and the whole human detrimentally affect that produces.
Below in conjunction with embodiment and accompanying drawing clone's process to Alocasia ordora agglutinant protein gene provided by the invention, gene function analysis, and application approach specifies.
Description of drawings:
Fig. 1 is the electrophoresis detection figure of 5 ' Race-PCR amplified production of AML protein gene.M represents the Marker of 2000bp among the figure, and swimming lane 1 is about 700bp for being 5 ' Race-PCR product of primer with W4236, and swimming lane 2 is about 600bp for being 5 ' Race-PCR product of primer with W4237.
W4236：5’-GCCGTTGCCGTGGGTGTTGGACTGCCA-3’ (27bp)
W4237：5’-TGCCGTCGCCGTAGAGGACCTG-3’ (22bp)
Fig. 2 is the electrophoresis detection figure of single double digestion product of 5 ' Race-PCR amplified production of AML protein gene and pMD 18-T vector carrier recombinant plasmid.M represents λ-HindIII digest among the figure.
1.1,1.2 be respectively the pUC18 empty plasmid and be the double digestion product with the single endonuclease digestion products of EcoRI, HindIII, 1.3
2.1,2.2 be respectively the recombinant plasmid 1 usefulness EcoRI among Fig. 4 .8, the enzyme of HindIII is cut product
2.3 be the double digestion product
3.1,3.2 be respectively the recombinant plasmid 2 usefulness EcoRI among Fig. 4 .8, the enzyme of HindIII is cut product
3.3 be the double digestion product
Fig. 3 is the electrophoresis detection figure of 3 ' Race-PCR amplified production of AML protein gene.M represents the Marker of 2000bp among the figure, and swimming lane 1 is about 600bp for being 3 ' Race-PCR product of primer with W2989, swimming lane 2 is for being 3 ' Race-PCR product of primer with W2986, be about 770bp, swimming lane 3 is about 800bp for being 3 ' Race-PCR product of primer with W2987.
W2986:5 '-ACTACGCCGCCGTCSTCCATCCGGA-3 ' is (S is G or C) (25bp)
W2987：5’-TCTACGGCCCATCCGTCTTCAAGA-3’ (24bp)
W2989：5’-TGGCAGTCCAACACCCACGGCAACG-3’ (25bp)
Fig. 4 is the electrophoresis detection figure of single double digestion product of 3 ' Race-PCR amplified production of AML protein gene and pMD 18-T vector carrier recombinant plasmid.M represents λ-HindIII digest among the figure.1.1,1.2 be respectively the pUC18 empty plasmid and be the double digestion product with the single endonuclease digestion products of EcoRI, HindHI, 1.3
2.1,2.2 enzymes that are respectively recombinant plasmid 2 usefulness EcoRI, HindIII among Fig. 4 .7 cut product, 2.3 for the double digestion product
3.1,3.2 enzymes that are respectively recombinant plasmid 3 usefulness EcoRI, HindIII among Fig. 4 .7 cut product, 3.3 for the double digestion product
Fig. 5 is the RACE-PCR product electrophorogram of AML full length gene sequence.M represents the Marker of 2000bp among the figure, and swimming lane 1,2 is about 1150bp for being two repetitions of 3 ' Race-PCR product of primer with W6045.
W6045：5’-GATCACGGCGAAGAAGAGGTGT-3’ (22bp)
Embodiment:
1, the clone of Alocasia ordora agglutinant protein gene (AML) 3 ' terminal sequence
The present invention sends out 3 ' terminal sequence of clone AML gene with RACE PCR.At first retrieve Genbank and pertinent literature, search the five kinds of lectins of other three kind of plant under the Araeceae with the nearly edge of Alocasia ordora or the nucleotide sequence and the aminoacid sequence of albuminoid, carry out arrangement analysis with five, find out three sections zones that wherein conservative property is high and be used for designing the essential gene-specific primer (GSP) of 3 ' RACE PCR, carry out 3 ' RACE pcr amplification then and obtain 3 ' terminal sequence fragment, again the PCR product is directly linked to each other with pMP 18-T vector carrier, be transformed in the competent escherichia coli cell, carry out the resistance screening recombinant plasmid by the LB flat board that contains AMP, select single bacterium colony shake-flask culture from flat board, carry out single double digestion reaction behind the extraction recombinant plasmid and add the electrophoresis detection, two kinds of enzymes using are respectively E10RI and HindIII, check order containing the segmental recombinant plasmid of purpose in the double digestion product.
2, the clone of AML gene 5 ' terminal sequence
The present invention uses 5 ' terminal sequence of RACE PCR method clone AML gene.Ultimate principle is identical with 3 ' end RACE PCR with operating process.But it only reacts at high two GSP of two sections zone design of conservative property.
3, the acquisition of the full length sequence of AML gene and clone
After finishing 3 ' and 5 ' order-checking, two terminal sequences are spliced merging by intermediary overlapping part, thereby obtain full length sequence.Choose 22 base synthetic primers from 5 ' end start-up portion, as the required gene-specific primer of RACE PCR of clone's full length sequence.RACE PCR product is carried out electrophoresis detection, and the result shows that length scale just in time conforms to the full length sequence that splicing obtains.
4,,, push over out aminoacid sequence from cDNA by with the translate in several different translation (instrument) softwares such as primer premier 5.0, DNA sis V2.5, the ExPASy website by the full length cDNA sequence of gained.The aminoacid sequence of Alocasia ordora lectin comprises 270 amino acid, comprising the structural domain of former and later two seminose specific combination, by Gln (glutamine), Asp (aspartic acid), Asn (l-asparagine), Val (Xie Ansuan), Tyr (tyrosine) five amino acid is formed.
Alocasia ordora agglutinant protein gene cDNA and AA sequence electronic file
＜110〉Agricultural University Of South China
＜120〉plain cDNA of Alocasia ordora lectin and AA sequence
<160>2
<210>1
<211>1124
<212>DNA
＜213〉Alocasia ordora (Alocasia macrorrhiza (L.) Schott)
<220>
<221>cDNA?sequence
<222>(1)...(1124)
<220>
<221>CDS
<222>(64)...(876)
<400>1
gatcacggcg aagaagaggt gttagggttt tgaatttagt agctagcagc aaccccattc 60
ctc?atg?gcc?aag?ctc?ctc?ctc?ttc?ctc?ctc?ccg?gcc?att?ctc?ggc?ctc?ctc?gtt?cct?cgg 120
Met?Ala?Lys?Leu?Leu?Leu?Phe?Leu?Leu?Pro?Ala?Ile?Leu?Gly?Leu?Leu?Val?Pro?Arg 19
tca?gcc?acg?gca?ata?ggc?atc?aac?tac?ctg?ctg?tcc?gga?gaa?acc?ctg?gac?acg?aac?ggc 180
Ser?Ala?Thr?Ala?Ile?Gly?Ile?Asn?Tyr?Leu?Leu?Ser?Gly?Glu?Thr?Leu?Asp?Thr?Asn?Gly 39
cat?ctc?agg?aac?ggc?aac?ttc?gac?ttg?gtc?atg?cag?gag?gac?tgc?aac?gcc?gtc?ctg?tac 240
His?Leu?Arg?Asn?Gly?Asn?Phe?Asp?Leu?Val?Met?Gln?Glu?Asp?Cys?Asn?Ala?Val?Leu?Tyr 59
aat?ggc?ggt?tgg?cag?tcc?aac?acg?gcc?aac?aga?gga?cga?gac?tgc?aag?ctc?tcc?ctg?acc 300
Asn?Gly?Gly?Trp?Gln?Ser?Asn?Thr?Ala?Asn?Arg?Gly?Arg?Asp?Cys?Lys?Leu?Ser?Leu?Thr 79
gac?tac?ggc?gaa?ctc?gtc?atc?aaa?aat?ggc?gac?gga?tcc?act?gtc?tgg?agg?agc?ggt?tcc 360
Asp?Tyr?Gly?Glu?Leu?Val?Ile?Lys?Asn?Gly?Asp?Gly?Ser?Thr?Val?Trp?Arg?Ser?Gly?Ser 99
cag?tcc?gac?aag?ggc?aag?tac?gcc?gcc?gtc?gtc?cat?ccg?gat?ggg?agg?ctg?gtc?gtc?tac 420
Gln?Ser?Asp?Lys?Gly?Lys?Tyr?Ala?Ala?Val?Val?His?Pro?Asp?Gly?Arg?Leu?Val?Val?Tyr 119
ggg?cca?tcc?gtc?ttt?aat?att?aat?ccc?tgg?gtc?ccc?ggc?ctc?aac?agc?ctg?cgg?ctc?ggc 480
Gly?Pro?Ser?Val?Phe?Asn?Ile?Asn?Pro?Trp?Val?Pro?Gly?Leu?Asn?Ser?Leu?Arg?Leu?Gly 139
aac?atc?ccc?ttc?aca?agc?aac?atg?ctc?ttc?tcc?gaa?caa?gtc?ctc?tac?gaa?gac?ggc?agg 540
Asn?Ile?Pro?Phe?Thr?Ser?Asn?Met?Leu?Phe?Ser?Glu?Gln?Val?Leu?Tyr?Glu?Asp?Gly?Arg 159
ctc?acc?gcg?aag?aac?cac?agg?ctc?gtc?atg?cag?ggc?gac?tgc?aac?ctg?gtc?cta?tac?ggt 600
Leu?Thr?Ala?Lys?Asn?His?Arg?Leu?Val?Met?Gln?Gly?Asp?Cys?Asn?Leu?Val?Leu?Tyr?Gly 179
ggc?aaa?ttc?ggc?tgg?cag?tcc?aac?acc?cac?ggc?aac?ggc?gag?gac?tgc?ttc?gtc?agg?ctc 660
Gly?Lys?Phe?Gly?Trp?Gln?Ser?Asn?Thr?His?Gly?Asn?Gly?Glu?Asp?Cys?Phe?Val?Arg?Leu 199
aac?cac?aag?ggc?gag?ctc?gtc?atc?aag?cac?gac?aac?ttc?agg?acc?atc?tgg?agc?agc?caa 720
Asn?His?Lys?Gly?Glu?Leu?Val?Ile?Lys?Asp?Asp?Asn?Phe?Arg?Thr?Ile?Trp?Ser?Ser?Gln 219
caa?aac?tcg?aat?gag?ggt?gac?tac?gtc?ttc?atc?ctc?cag?gac?gac?ggc?ttc?ggc?gtc?atc 780
Gln?Asn?Ser?Asn?Glu?Gly?Asp?Tyr?Val?Phe?Ile?Leu?Gln?Asp?Asp?Gly?Phe?Gly?Val?Ile 239
tac?ggc?cct?gcc?atc?tgg?gcg?acc?agc?tcg?aag?cgc?tcc?att?gct?gcg?gag?gag?aag?atg 840
Tyr?Gly?Pro?Ala?Ile?Trp?Ala?Thr?Ser?Ser?Lys?Arg?Ser?Ile?Ala?Ala?Glu?Glu?Lys?Met 259
agc?ggc?atg?gtg?cct?gag?aac?gtg?gaa?gcg?aaa?taa 876
Ser?Gly?Met?Val?Pro?Glu?Asn?Val?Glu?Ala?Lys 270
agtt ggcaactata tatcttccgc 900
ctcgctagaa aaataaatga gcgcatgaca tgttgctcca tgctagctag tttctggacc 960
gtaaaataat cgtgtgcagt acgtgcgatt attcgctgtt gtagtctgtc ttcgtcgtag 1020
agtgttctta attaagaggc tttcggccgt tgctttcgta gccataggtc tggctttttg 1080
ctggtgtaaa atggtccttt gaataaatat ggctaccttg agtt 1124
<210>2
<211>270
<212>protein
＜213〉Alocasia ordora (Alocasia macrorrhiza (L.) Schott)
<400>2
Met?Ala?Lys?Leu?Leu?Leu?Phe?Leu?Leu?Pro?Ala?Ile?Leu?Gly?Leu?Leu?Val?Pro?Arg 19
Ser?Ala?Thr?Ala?Ile?Gly?Ile?Asn?Tyr?Leu?Leu?Ser?Gly?Glu?Thr?Leu?Asp?Thr?Asn?Gly 39
His?Leu?Arg?Asn?Gly?Asn?Phe?Asp?Leu?Val?Met?Gln?Glu?Asp?Cys?Asn?Ala?Val?Leu?Tyr 59
Asn?Gly?Gly?Trp?Gln?Ser?Asn?Thr?Ala?Asn?Arg?Gly?Arg?Asp?Cys?Lys?Leu?Ser?Leu?Thr 79
Asp?Tyr?Gly?Glu?Leu?Val?Ile?Lys?Asn?Gly?Asp?Gly?Ser?Thr?Val?Trp?Arg?Ser?Gly?Ser 99
Gln?Ser?Asp?Lys?Gly?Lys?Tyr?Ala?Ala?Val?Val?His?Pro?Asp?Gly?Arg?Leu?Val?Val?Tyr?119
Gly?Pro?Ser?Val?Phe?Asn?Ile?Asn?Pro?Trp?Val?Pro?Gly?Leu?Asn?Ser?Leu?Arg?Leu?Gly?139
Asn?Ile?Pro?Phe?Thr?Ser?Asn?Met?Leu?Phe?Ser?Glu?Gln?Val?Leu?Tyr?Glu?Asp?Gly?Arg?159
Leu?Thr?Ala?Lys?Asn?His?Arg?Leu?Val?Met?Gln?Gly?Asp?Cys?Asn?Leu?Val?Leu?Tyr?Gly?179
Gly?Lys?Phe?Gly?Trp?Gln?Ser?Asn?Thr?His?Gly?Asn?Gly?Glu?Asp?Cys?Phe?Val?Arg?Leu?199
Asn?His?Lys?Gly?Glu?Leu?Val?Ile?Lys?Asp?Asp?Asn?Phe?Arg?Thr?Ile?Trp?Ser?Ser?Gln?219
Gln?Asn?Ser?Asn?Glu?Gly?Asp?Tyr?Val?Phe?Ile?Leu?Gln?Asp?Asp?Gly?Phe?Gly?Val?Ile?239
Tyr?Gly?Pro?Ala?Ile?Trp?Ala?Thr?Ser?Ser?Lys?Arg?Ser?Ile?Ala?Ala?Glu?Glu?Lys?Met?259
Ser?Gly?Met?Val?Pro?Glu?Asn?Val?Glu?Ala?Lys 270
Claims (3)
1, the Alocasia ordora agglutinant protein gene that obtains of a kind of separation is characterized in that:
(1) has the nucleotide sequence of sequence 1 in the sequence table, wherein the 1st～63 bit base is for containing the sequence in promoter regulation element and the non-lectin translation of 5 ' end district, the 64th～873 bit base is opening code-reading frame (ORF), and the 874th～1124 bit base is the sequence in the non-lectin translation of 3 ' end district;
(2) the amino acid polypeptide sequence shown in the sequence 2 in the code sequence tabulation, this sequence is made up of 270 amino-acid residues, the 3 d function structural domain of two seminose specific combination before and after having;
(3) have sequence 1 in the series of tables nucleotide sequence or with the 64th～873 bit base sequence of sequence 1 in continuous 100 zones more than the base homology more than 95% is arranged, have the nucleotide sequence of the equal functional protein of coding and sequence 2;
(4) the amino acid polypeptide sequence of coding Alocasia ordora lectin precursor protein, this polypeptide has the sequence shown in the sequence 2 in the sequence table; Or coding have same function by sequence 2 polypeptides derived sequences, or its conservative property variation polypeptide, or its active fragments.
2, according to the said gene of claim 1, it is characterized in that coded amino acid polypeptide sequence contains the functional domain of being made up of 5 PPR of amino-acid residue repeating unit, this structural domain is by Gln (glutamine Q), Asp (aspartic acid D), Asn (l-asparagine N), Val (Xie Ansuan V), Tyr (tyrosine Y) five amino acid constitutes, five wherein previous AA positions are respectively: Gln51, Asp53, Asn55, Val57, Tyr59, back one is: Gln170, Asp172, Asn174, Val176, Tyr178.
3, the Alocasia ordora agglutinant protein gene that obtains of a kind of separation is characterized in that: with gene or contain carrier converting cotton or other plant of gene, cultivate and can explain the genetically modified crops of said gene, thereby have pest-resistant effect.
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| CNA031399576A CN1537943A (en) | 2003-07-28 | 2003-07-28 | Alocasia odora agglutinin protein gene and its application |
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| CNA031399576A CN1537943A (en) | 2003-07-28 | 2003-07-28 | Alocasia odora agglutinin protein gene and its application |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2007012277A1 (en) * | 2005-07-26 | 2007-02-01 | The Chinese University Of Hong Kong | Isolated proteins from a traditional chinese medicine yuzhu and use thereof |
| CN102952183A (en) * | 2011-08-17 | 2013-03-06 | 中国农业科学院生物技术研究所 | Protein related to plant aphid resistance, and coding gene and application thereof |
| CN107936100A (en) * | 2017-11-17 | 2018-04-20 | 四川大学 | Common calanthe herb mannose-binding protein |
-
2003
- 2003-07-28 CN CNA031399576A patent/CN1537943A/en active Pending
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2007012277A1 (en) * | 2005-07-26 | 2007-02-01 | The Chinese University Of Hong Kong | Isolated proteins from a traditional chinese medicine yuzhu and use thereof |
| US7378513B2 (en) | 2005-07-26 | 2008-05-27 | The Chinese University Of Hong Kong | Isolated proteins from a traditional Chinese medicine Yuzhu and use thereof |
| CN1974598B (en) * | 2005-07-26 | 2010-05-26 | 香港中文大学 | Isolated proteins from a traditional Chinese medicine yuzhu and use thereof |
| CN102952183A (en) * | 2011-08-17 | 2013-03-06 | 中国农业科学院生物技术研究所 | Protein related to plant aphid resistance, and coding gene and application thereof |
| CN102952183B (en) * | 2011-08-17 | 2014-11-05 | 中国农业科学院生物技术研究所 | Protein related to plant aphid resistance, and coding gene and application thereof |
| CN107936100A (en) * | 2017-11-17 | 2018-04-20 | 四川大学 | Common calanthe herb mannose-binding protein |
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