CN102505021A - Soluble acid invertase (So INV) gene of sugarcane GH32 family and protein sequence thereof - Google Patents
Soluble acid invertase (So INV) gene of sugarcane GH32 family and protein sequence thereof Download PDFInfo
- Publication number
- CN102505021A CN102505021A CN2011103657488A CN201110365748A CN102505021A CN 102505021 A CN102505021 A CN 102505021A CN 2011103657488 A CN2011103657488 A CN 2011103657488A CN 201110365748 A CN201110365748 A CN 201110365748A CN 102505021 A CN102505021 A CN 102505021A
- Authority
- CN
- China
- Prior art keywords
- ala
- gly
- leu
- val
- thr
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Landscapes
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The invention discloses a soluble acid invertase (So INV) gene of the sugarcane GH32 family and a protein sequence thereof. On the basis of analyzing the full length of the So INV gene sequences of all plants and by taking the conserved region of So INV gene sequences of the same families to design the prime, total RNA (ribose nucleic acid) is extracted from young leaves of sugarcane; the total RNA is processed with reverse transcription and is amplified through a conventional PCR (Polymerase Chain Reaction); method and the total RNA is cloned to total length of the c DNA (deoxyribonucleic acid) sequence of the three members of the sugarcane So INV gene family through combining a RACE PCR (rapid-amplification of c DNA ends) technology. The invention lays a foundation for the study of transcription and expression mechanism of sugarcane So INV as well as for the further study of accumulation mechanism of sugarcane; with the amino acid sequence, purified protein with bioactivity can be obtained; the invention overcomes the technical difficulty that the sugarcane INV gene of the glycosyl hydrolase family GH32 gene family so far only clones a few c DNA sequence fragments and cannot obtain the total length of the nucleotide sequence and provides a foundation for the study of the biological function of the So INV.
Description
Technical field
The present invention relates to a kind of sugarcane GH32 family's soluble acid invertase (SoINV) gene and protein sequence thereof.
Background technology
Soluble acid invertase (Soluble Acid Invertase) (hereinafter to be referred as INV) is the key enzyme of higher plant body inner control Sucrose Metabolism, and irreversible catalysis sucrose cracking forms glucose and fructose.Mainly be present in the vacuole, in the process of regulation and control hexose and sucrose level, play a significant role, and closely related with fruit development, maturation and sugared accumulation.
Different plant seedlings, spire, young root, young fruit are being discovered highly active INV is often relevant with expanding rapidly of the growth of fruit or storage organ.Lingle research shows that in sweet sorghum stalk, the activity of SAI enzyme and the length of internode have very high positive correlation, and Li Xingjun studies during to the red bayberry flower bud initiation, and the discovery sucrose content becomes remarkable negative correlation property with the work of Inv enzyme.INV mainly is responsible for the degraded of sucrose in the vacuole, and its active shortage is that sucrose begins the cumulative prerequisite.Plant that sugar degree is high such as sweet sorghum, beet, citrus, sugarcane all have the active characteristics of the acid Inv of shortage.When sweeter and sweetless cucurbitaceous plant fruit sucrose accumulation characteristic, find that the sucrose of input maybe be by the district at a distance from vacuole in sweetless fruit, sucrose decomposition becomes hexose under the effect of INV.Further the two genotypic difference of research is not to be to import and distinguish separated ability but the further metabolism of the sucrose that is to import, and the shortage of INV can hinder the Sucrose Metabolism in this step, and sucrose then accumulates subsequently, and fruit sweetens.
The Inv vigor has expression specificity at the different steps and the different tissues organ of growth and development of plants.Yam Inv expression amount in the seed of the tender source organ of children such as leaf, root, sprouting is very high, and expression amount is seldom in the cell of storehouse.Handle with GA, can improve that acid Inv is active to reduce sucrose level, cause the prosperous length of young sprout and consume reducing sugar through inducing.The Inv expression of gene also receives the adjusting of sugar, ethene, ABA simultaneously, and concrete regulation mechanism also is not very clear.
The active rising of INV receives the control of mRAN level in the tamato fruit ripening process, and to compare with the GUS transfer-gen plant with the non-transgenic plant be low-down to the vigor of INV in the tamato fruit that it is changed over to antisense INV gene plant.And it is extremely low to the sucrose content in the non-transgenic tamato fruit; But then significantly improve at all content that have the transfer-gen plant sucrose of inverted defined gene; The content of hexose descends, and shows that non-transgenic tomato highly active INV in mellow fruit has stoped the accumulation of sucrose.Receive effective inhibition although have the transgenic Fructus Lycopersici esculenti INV activity of inverted defined gene, these transgenic Fructus Lycopersici esculentis still accumulate a certain amount of hexose in full ripe stage.
Exist the isozyme form of multiple saccharase in the higher plant tissue.Can be divided into two big types according to ph optimum: acid invertase and neutrality/alkaline saccharase; Can be divided three classes in the residing position of cell according to it: cell walls saccharase, vacuolus converzyme and tenuigenin saccharase.Cell walls saccharase and vacuolus converzyme all are partial to acidity owing to ph optimum, therefore lump together to belong to the acid invertase class.All acid invertases all are glycosylated, the hydrolysis that they not only can catalysis sucrose, and can some oligosaccharides of catalysis such as the hydrolysis of raffinose and stachyose.Glycosyl hydrolase (glycosyl hydrolase familie) is divided into four gene family: GH32, GH43, GH62, GH68.Sugarcane soluble acid invertase of the present invention (SoINV) gene belongs to GH32 family.Up to the present sugarcane has only cloned some INV cDNA sequence fragments, and does not have the further report of research.
Summary of the invention
The objective of the invention is: function and application sugarcane GH32 family's soluble acid invertase (SoINV) genetically modified crops kind for further studying sugarcane GH32 family's soluble acid invertase (SoINV) gene provide a kind of sugarcane GH32 family's soluble acid invertase (SoINV) gene and protein sequence thereof.
The technical scheme that the present invention taked is following:
A kind of sugarcane GH32 family's soluble acid invertase (SoINV) gene and protein sequence thereof; Be that all plant soluble acid invertase full length gene sequences are carried out on the basis of evolutionary analysis; Compare the soluble acid invertase gene cDNA sequence of same family, and the segmental primer of design amplification SoINV gene core, total RNA extracted from the sugarcane young leaflet tablet; And through reverse transcription, with conventional PCR method amplification SoINV gene core fragment; Some in core fragment sequence 5 ' end design then to special primer; At core fragment sequence a 3 ' end special primer of design and two anchor primers; Through RACE round pcr 5 of SoINV gene core district ' three different sequence fragments of end and the 3 ' sequence fragment of end that increases respectively; Three 5 ' terminal sequences are spliced with core area, 3 ' terminal sequence and AY302083 fragment respectively; Obtain sugarcane GH32 three soluble acid invertases of family (SoINV) full length gene cDNA sequence, be designated as SoINV1, SoINV2 and SoINV3 respectively.
Above-described SoINV1 length overall 2387bp; Through Vector NTI Advance 11 software analysis, the ORF of this sequence is 2055bp, 685 amino acid of encoding; Initiator codon (ATG) is positioned at 215bp place behind the transcription initiation site, and terminator codon (TAG) is positioned at 2272bp, also has the non-coding sequence of one section 115bp thereafter, and has the typical polyA tail of eukaryote; The proteic aminoacid sequence of the sugarcane GH32 SoINV1 of family coded by said gene:
METRDTTAPLPYSYTPLPAADAASAEVTGTGGRSRRRSLCAAALVLSAALLLAVAALAAAGRRPTTAVGETAGVGVVPGVGTPQATSTRSISRGPDAGVSEKTSGAWSGVVDDGGRLRADGGGNAFPWSNAMLQWQRTGFHFQPQRNWMNDPNGPVYYKGWYHLFYQYNPDGAIWGNKIAWGHAVSRDLIHWRHLPLAMLPDQWYDTNGVWTGSATTLPDGRLAMLYTGSTNTSVQVQCLAVPADDDDPLLTNWTKYEGNPALYPPPGIGPRDFRDPTTAWFDPSDSTWRIVIGSKDDAEGDHAGIAVVYRTRDFVHFELLPDLLHRVAGTGMWECIDFYPVATRGKASGNGVDMSDALAKNGAVVGDVVHVMKASMDDDRHDYYALGRYDAAANAWTPLDAEKDVGTGLRYDWGKFYASKTFYDPAKRRRVLWGWVGETDSERADVSKGWASLQGIPRTVLLDTKTGSNLLQWPVEEVE?TLRTNSTDLSGITIDYGSTFPLNLRRATQLDIEAEFELDRRAVMSLNEADVGYNCSTSGGAAARGALGPFGLLVLTDKHLHEQTAVYFYVAKGLDGSLTTHFCQDESRSSSANDIVKRVVGSAVPVLEDETTLSLRVLVDHSIVESFAQGGRSTATSRVYPTKAIYANAGVFLFNNATAARVTAKKLVVHEMDSSYNHDYMVTDI
Above-described SoINV2 length overall 2429bp; Through Vector NTI Advance 11 software analysis, the ORF of this sequence is 2085bp, 695 amino acid of encoding; Initiator codon (ATG) is positioned at 227bp place behind the transcription initiation site, and terminator codon (TAA) is positioned at 2314bp, also has the non-coding sequence of one section 115bp thereafter, and has the typical polyA tail of eukaryote; The proteic aminoacid sequence of the sugarcane GH32 SoINV2 of family coded by said gene:
METRDTTAPLPYSYTPLPAADAASAEVTGTGHRGGGRSRRSSLCAAALVLSAALLLAVAALAGVGGRVAVVPRPTTAVGETAGVGVGPGAGTPQATSTRSISRGPDAGVSEKTSGAWSGVVDDGGRLRADGGGNAFPWSNAMLQWQRTGFHFQPQRNWMNDPNGPVYYKGWYHLFYQYNPDGAIWGNKIAWGHAVSRDLIHWRHLPLAMLPDQWYDTNGVWTGSATTLPDGRLAMLYTGSTNTSVQVQCLAVPAGDDDPLLTNWTKYEGNPALYPPPGIGPRDFRDPTTAWFDPSDSTWRIVIGSKDDAEGDHAGIAVVYRTRDFVHFELLPDLLHRVAGTGMWECIDFYPVATRGKASGNGVDMSDALAKNGAVVGDVVHVMKASMDDDRHDYYALGRYDAAANAWTPLDAEKDVGTGLRYDWGKFYASKTFYDPAKRRRVLWGWVGETDSERADVSKGWASLQGIPRTVLLDTKTGSNLLQWPVEEVETLRTNSTDLSGITIDYGSTFPLNLRRATQLDIEAEFELDRRAVMSLNEADVGYNCSTSGGAAARGALGPFGLLVLTDKHLHEQTAVYFYVAKGLDGSLTTHFCQDESRSSSANDIVKRVVGSAVPVLEDETTLSLRVLVDHSIVESFAQGGRSTATSRVYPTKAIYANAGVFLFNNATAARVTAKKLVVHEMDSSYNHDYMVTDI
Above-described SoINV3 length overall 2373bp; Through Vector NTI Advance 11 software analysis, the ORF of this sequence is 2088bp, 696 amino acid of encoding; Initiator codon (ATG) is positioned at 168bp place behind the transcription initiation site, and terminator codon (TAA) is positioned at 2258bp, also has the non-coding sequence of one section 115bp thereafter, and has the typical polyA tail of eukaryote; The proteic aminoacid sequence of the sugarcane GH32 SoINV3 of family coded by said gene is:
METRDTTAPLPYSYTPLPAADAASAEVTGTGGSRSRRRRPLCAAALVLSAALLLAVAALAGVGSRVAAVVPRPTTAVGETAGVGVGVVPGAGTPQATSTRSRSRGPDAGVSEKTSGVWTGVIDDGARLRTDAGGNAFPWSNAMLQWQRTGFHFQPQRNWMNDPNGPVYYKGWYHLFYQYNPDGAIWGNKIAWGHAVSRDLIHWRHLPLAMLPDQWYDTNGVWTGSATTLPDGRLAMLYTGSTNTSVQVQCLAVPADDDDPLLTNWTKYEGNPALYPPPGIGPRDFRDPTTAWFDPSDSTWRIVIGSKDDAEGDHAGIAVVYRTRDFVHFELLPDLLHRVAGTGMWECIDFYPVATRGKASGNGVDMSDALAKNGAVVGDVVHVMKASMDDDRHDYYALGRYDAAANAWTPLDAEKDVGTGLRYDWGKFYASKTFYDPAKRRRVLWGWVGETDSERADVSKGWASLQGIPRTVLLDTKTGSNLLQWPVEEVETLRTNSTDLSGITIDYGSTFPLNLRRATQLDIEAEFELDRRAVMSLNEADVGYNCSTSGGAAARGALGPFGLLVLTDKHLHEQTAVYFYVAKGLDGSLTTHFCQDESRSSSANDIVKRVVGSAVPVLEDETTLSLRVLVDHSIVESFAQGGRSTATSRVYPTKAIYANAGVFLFNNATAARVTAKKLVVHEMDSSYNHDYMVTDI
Above-described sugarcane GH32 three soluble acid invertases of family (SoINV) full length gene cDNA sequence, core area sequence (M-SoINV) is to use following nucleotide sequence to obtain as primer amplification:
N?f1：5′-tctggggcaacaagatcgcgt-3′；
N?r1：5′-aaattgggtgcagcggtgggt-3′。
The sequence of above-described 3 ' end (3 '-SoINV) be to use following nucleotide sequence to be primer, with 3 '-primer that Full RACEKit (TAKARA) provides combines, the acquisition of increasing of Bird's Nest formula:
GZ?f2：5′-cctcttcaacaacgccaccgccg-3′。
Above-described 5 ' terminal sequence (5 '-SoINV) be respectively to use following nucleotide sequence to be primer, with 5 '-primer that Full RACEKit (TAKARA) provides combines, and the amplification of Bird's Nest formula obtains:
The primer of the soluble acid invertase SoINV1 of sugarcane GH32 family gene 5 ' end is used to increase:
INVSYR1：(5′-gttggtggagccggtgtagagcat-3′)；
INVR1：(5′-gcccctgctgatgctcctggtcg-3′)；
The primer of the soluble acid invertase SoINV2 of sugarcane GH32 family gene 5 ' end is used to increase:
INVSY?R1：(5′-gttggtggagccggtgtagagcat-3′)；
INVR2：(5′-acgtcgcctgtggtgtcccc-3′)；
The primer of the soluble acid invertase SoINV3 of sugarcane GH32 family gene 5 ' end is used to increase:
INVSY?R1：(5′-gttggtggagccggtgtagagcat-3′)；
INVR3：(5′-ggggtcgttcatccagttcctct-3′)。
Advantage of the present invention and effect thereof:
A kind of sugarcane GH32 of the present invention family's soluble acid invertase (SoINV) gene order; Be not merely the transcript and expression mechanism of research sugarcane soluble acid invertase; The accumulation mechanism of further inquiring into sucrose lays the foundation; And can obtain the purifying protein of biologically active through its aminoacid sequence, for the biological function of research soluble acid invertase and utilize the SoINV of GH32 family gene to carry out the sugar cane breed improvement basis is provided.
Description of drawings
Fig. 1: the full length cDNA sequence of sugarcane GH32 family soluble acid invertase (SoINV1) gene;
Fig. 2: the proteic aminoacid sequence of sugarcane GH32 family soluble acid invertase (SoINV1) coded by said gene;
Fig. 3: the full length cDNA sequence of sugarcane GH32 family soluble acid invertase (SoINV2) gene;
Fig. 4: the proteic aminoacid sequence of sugarcane GH32 family soluble acid invertase (SoINV2) coded by said gene;
Fig. 5: the full length cDNA sequence of sugarcane GH32 family soluble acid invertase (SoINV3) gene;
Fig. 6: the proteic aminoacid sequence of sugarcane GH32 family soluble acid invertase (SoINV3) coded by said gene.
Embodiment
Embodiment 1 (the sugarcane GH32 SoINV1 of family)
1, plant INV gene evolution is analyzed: with " soluble acid invertase " search NCBI website (http://www.ncbi.nlm.nih.gov) nonredundancy pool of amino acids, obtain all plant INV sequences.All sequences to being obtained is analyzed, and removes repeating sequences.Obtain 20 of plant INV sequences altogether, wherein full length sequence is 7.With ClustalX the aminoacid sequence of all total length INV is compared, and comparison result is saved as the PHYLIP form; Use the Seqboot of PHYLIP software package then, Protdi st, Neighbor and Consense program are carried out evolutionary analysis, and all total length INV genes of plant are divided into corresponding family.
2, the extraction of sugarcane RNA:
(1) gets fresh cane lobus cardiacus (osmanthus sugar 28), in liquid nitrogen, fully be ground into powder.Take by weighing the product after 100mg grinds, put into 1.5ml centrifuge tube, add day TRNzol-A of root biochemical technology ltd production with the liquid nitrogen precooling
+Extracting solution 1ml;
(2) with the abundant mixing of sample, room temperature held 5min makes the nucleic acid-protein mixture separate fully;
(3) control is 4 ℃, and 14,000g, centrifugal 10min;
(4) draw supernatant in another 1.5ml centrifuge tube, adding the chloroform volume is 1/5 of supernatant volume, the mixing 15sec that fully vibrates, and room temperature is placed 2～3min;
(5) control is 4 ℃, and 14,000g, centrifugal 15min;
(6) repeating step 4,5 once;
(7) draw the upper strata aqueous phase solution in another 1.5ml centrifuge tube, add the Virahol of equal-volume precooling, mixing 10min places 25min on ice;
(8) control is 4 ℃, and 14,000g behind the centrifugal 10min, forms white gelatinous precipitate at the pipe side and the pipe end;
(9) remove supernatant, stay deposition.Add 1ml 75% washing with alcohol deposition 2 times (each 4 ℃, 2,300g, centrifugal 3min);
(10) pour out liquid, room temperature is placed 3-5min.Add 30-50 μ l DEPC treating water, fully the dissolving back is in-70 ℃ of preservations;
(11) get 1 μ l RNA sample and carry out electrophoresis detection with 1% sepharose, other gets 1 μ l RNA sample and surveys the OD value.
3, RT-PCR (reverse transcription PCR):
(1) centrifuge tube of a 0.2ml of preparation adds following composition:
The total RNA:0.1-5 μ of sugarcane g;
Oligo (dT) 18 primers (0.5 μ g/ μ l): 1 μ l;
Add DEPC-treated water to TV: 12 μ l.
(2) slight mixing, brief centrifugal after, 65 ℃ of incubation 5min place cooled on ice.
(3) add following composition on request:
5×Reaction?Buffer： 4μl；
RiboLock
TM?RNase?Inhibitor(20u/μl)： 1μl；
10mM?dNTP?Mix： 2μl；
ReverAid
TM?M-MuLV?Reverse?Tranxcriptase：1μl；
TV is 20 μ l.
(4) slight mixing is briefly centrifugal.
(5) 42 ℃ of incubation 60min.
(6) 70 ℃ of incubation 5min termination reactions ,-20 ℃ of preservations are subsequent use.
4, SoINV gene intermediate segment clone: the cDNA sequence of comparison monocotyledons INV gene on Vector NTI Advance 11, at the primer of a pair of SoINV intermediate segment that is used to increase of its conserved regions design.Carry out conventional pcr amplification with Nf1 and Nr1 primer, the PCR product reclaims the purifying rear clone to T carrier and order-checking through glue.Sequencing result shows, PCR product length 1601bp (M-SoINV).
Nf1：5′-tctggggcaacaagatcgcgt-3′；
Nr1：5′-aaattgggtgcagcggtgggt-3′。
5, SoINV gene 3 '-RACE clone: according to the intermediate segment sequence that a last pacing preface obtains, designed 3 '-gene specific primer of RACE.
GZ?f2：5′-cctcttcaacaacgccaccgccg-3′。
3 '-anchor primer that Full RACE Kit (TAKARA) provides:
GZ?r1：5′-ggccacgcgtcgactagtac-3′；
GZ?r2：5′-ggccacgcgtcgactagtacttttttttttttttt-3′。
Extract the total RNA of sugarcane young tender leaf, according to above-mentioned RT-PCR method, special primer GZr2 is the reverse transcription primer, and rt synthesizes cDNA.Carry out conventional pcr amplification with primer GZ f2 and GZ r1.Product reclaims the purifying rear clone to T carrier and order-checking through glue.Sequencing result shows, product length 218bp (3 '-SoINV).
6, SoINV gene 5 '-the RACE clone: with reference to intermediate segment and gene pool AY302083 gene upstream sequence, design two 5 '-gene specific primer of RACE,
Two 5 of SoINV1 genes '-gene specific primer of RACE:
INVSYR1：5′-gttggtggagccggtgtagagcat-3′；
INVR1：5′-gcccctgctgatgctcctggtcg-3′。
5 '-primer that Full RACE Kit (TAKARA) test kit provides:
Outer?Primer：5′-catggctacatgctgacagccta-3；
Inner?Primer：5′-cgcggatccacagcctactgatgatcagtcgatg-3′。
Extract the total RNA of sugarcane young tender leaf, according to 5 '-Full RACE Kit (TAKARA) operational manual carries out 5 '-RACE.Bird Testis type PCR: outside primers INVSYR1 and Outer? Primer PCR product as a template, and then less and InnerPrimer primer INVR1 second round of PCR, PCR products were gel purified and cloned into T vector and sequenced.Sequencing result shows, obtain 5 '-RACE product length be 496bp (5 '-SoINV1).
7, the soluble acid invertase SoINV1 of sugarcane GH32 family full length gene nucleotide sequence analysis:
With three sequences that obtained (5 '-SoINV1, M-SoINV and 3 '-SoINV) 0 splices with the AY302083 fragment, obtained soluble acid invertase (SoINV1) full length gene cDNA sequence, length overall 2387bp.Through Vector NTIAdvance 11 software analysis, the ORF of this sequence is 2055bp, 685 amino acid of encoding.Initiator codon (ATG) is positioned at 215bp place behind the transcription initiation site, and terminator codon (TAA) is positioned at 2272bp, also has the non-coding sequence of one section 115bp thereafter, and has the typical polyA tail of eukaryote.
Sugarcane GH32 family soluble acid invertase SoINV1 gene and protein sequence table see sequence table 1 for details.Embodiment 2 (the sugarcane GH32 SoINV2 of family)
1 to 5 step is with embodiment 1.
6, SoINV gene 5 '-the RACE clone: with reference to intermediate segment and gene pool AY302083 gene upstream sequence, design two 5 '-gene specific primer of RACE,
Two 5 of SoINV2 genes '-gene specific primer of RACE:
INVSYR1：(5′-gttggtggagccggtgtagagcat-3′)；
INVR2：(5′-acgtcgcctgtggtgtcccc-3′)。
5 '-primer that Full RACE Kit (TAKARA) test kit provides is with embodiment 1.
Extract the total RNA of sugarcane young tender leaf, according to 5 '-Full RACE Kit (TAKARA) operational manual carries out 5 '-RACE.Bird Testis type PCR: outside primers INVSYR1 and Outer? Primer PCR product as a template, and then less and InnerPrimer primer INVR2 second round of PCR, PCR products were gel purified and cloned into T vector and sequenced.Sequencing result shows, obtain 5 '-RACE product length be 515bp (5 '-SoINV2).
7, the soluble acid invertase SoINV2 of sugarcane GH32 family full length gene nucleotide sequence analysis:
With three sequences that obtained (5 '-SoINV2, M-SoINV and 3 '-SoINV) 0 splices with the AY302083 fragment, obtained SoINV2 full length gene cDNA sequence, length overall 2429bp.Through Vector NTI Advance 11 software analysis, the ORF of this sequence is 2085bp, 695 amino acid of encoding.Initiator codon (ATG) is positioned at 227bp place behind the transcription initiation site, and terminator codon (TAA) is positioned at 2314bp, also has the non-coding sequence of one section 115bp thereafter, and has the typical polyA tail of eukaryote.
Sugarcane GH32 family soluble acid invertase SoINV2 gene and protein sequence table see sequence table 2 for details.
Embodiment 3 (the sugarcane GH32 SoINV3 of family)
1 to 5 step is with embodiment 1.
6, SoINV gene 5 '-the RACE clone: with reference to intermediate segment and gene pool AY302083 gene upstream sequence, design two 5 '-gene specific primer of RACE,
Two 5 of SoINV3 genes '-gene specific primer of RACE:
INVSYR1：(5′-gttggtggagccggtgtagagcat-3′)；
INVR3：(5′-ggggtcgttcatccagttcctct-3′)。
5 '-primer that Full RACE Kit (TAKARA) test kit provides is with embodiment 1.
Extract the total RNA of sugarcane young tender leaf, according to 5 '-Full RACE Kit (TAKARA) operational manual carries out 5 '-RACE.Bird Testis type PCR: outside primers INVSYR1 and Outer? Primer PCR product as a template, and then within the primer INVR1, INVR2, INVR3 respectively, and the Inner? Primer for the second round of PCR, PCR products were gel purified and cloned into T vector and sequenced. .Sequencing result shows, obtain 5 '-RACE product length be respectively 496bp (5 '-SoINV1), 515bp (5 '-SoINV2) and 656bp (5 '-SoINV3).
7, the soluble acid invertase SoINV3 of sugarcane GH32 family full length gene nucleotide sequence analysis:
With three sequences that obtained (5 '-SoINV3, M-SoINV and 3 '-SoINV) splice with the AY302083 fragment, obtained SoINV3 full length gene cDNA sequence, length overall 2373bp.Through Vector NTI Advance 11 software analysis, the ORF of this sequence is 2088bp, 696 amino acid of encoding.Initiator codon (ATG) is positioned at 168bp place behind the transcription initiation site, and terminator codon (TAA) is positioned at 2258bp, also has the non-coding sequence of one section 115bp thereafter, and has the typical polyA tail of eukaryote.
Sugarcane GH32 family soluble acid invertase SoINV3 gene and protein sequence table see sequence table 3 for details.
Sequence table
Sequence table 1
< 110>Guangxi University
< 120>sugarcane soluble acid invertase (SoINV1) gene and protein sequence
<160>2
<210>1
<211>2387
<212>DNA
< 213>sugarcane (Saccharum officinarum)
<220>
<221>CDS
<222>(215)...(2272)
<220>
<221>5′UTP
<222>(1)...(214)
<220>
<221>3′UTP
<222>(2273)...(2387)
<400>1
GAAAAATGTG?TTGTTGTATG?TACTACTATA?TTCTAGTTAC?AATCGAGTCG?AATCGCCGCA?60
TTGCGCGGCT?CCAGATCGAA?TCGGCCGGCG?AGGAGTCGGT?CATCGTCGCT?CCGGCCGCCG?120
CCGTAGCCGC?CCCTAGAGAG?GGTCGCCCGC?CGTCGTAACC?GCAACACAAG?TCGCCGGCGG?180
CCTCCTTCCG?ATCGATCCTC?TTCCGCCGTC?GGCA?214
ATG?GAG?ACC?CGG?GAC?ACG?ACG?GCG?CCG?CTC?CCC?TAC?TCG?TAC?ACG?259
Met?Glu?Thr?Arg?Asp?Thr?Thr?Ala?Pro?Leu?Pro?Tyr?Ser?Tyr?Thr
5 10 15
CCG?CTG?CCG?GCC?GCC?GAC?GCC?GCG?TCC?GCC?GAG?GTC?ACC?GGC?ACC?304
Pro?Leu?Pro?Ala?Ala?Asp?Ala?Ala?Ser?Ala?Glu?Val?Thr?Gly?Thr
20 25 30
GGC?GGC?AGG?AGC?AGG?CGG?AGG?TCC?CTC?TGC?GCC?GCG?GCG?CTC?GTC?349
Gly?Gly?Arg?Ser?Arg?Arg?Arg?Ser?Leu?Cys?Ala?Ala?Ala?Leu?Val
35 40 45
CTC?TCC?GCC?GCG?CTG?CTC?CTC?GCC?GTG?GCC?GCG?CTC?GCC?GCC?GCC?394
Leu?Ser?Ala?Ala?Leu?Leu?Leu?Ala?Val?Ala?Ala?Leu?Ala?Ala?Ala
50 55 60
GGC?CGC?CGC?CCA?ACG?ACC?GCG?GTG?GGA?GAA?ACG?GCC?GGC?GTC?GGC?439
Gly?Arg?Arg?Pro?Thr?Thr?Ala?Val?Gly?Glu?Thr?Ala?Gly?Val?Gly
65 70 75
GTC?GTC?CCT?GGC?GTG?GGG?ACA?CCA?CAG?GCG?ACG?TCG?ACC?AGG?AGC?484
Val?Val?Pro?Gly?Val?Gly?Thr?Pro?Gln?Ala?Thr?Ser?Thr?Arg?Ser
80 85 90
ATC?AGC?AGG?GGC?CCC?GAC?GCC?GGC?GTG?TCG?GAG?AAG?ACG?TCC?GGC?529
lle?Ser?Arg?Gly?Pro?Asp?Ala?Gly?Val?Ser?Glu?Lys?Thr?Ser?Gly
95 100 105
GCG?TGG?AGC?GGC?GTC?GTC?GAC?GAT?GGC?GGG?AGG?CTC?CGT?GCT?GAC?574
Ala?Trp?Ser?Gly?Val?Val?Asp?Asp?Gly?Gly?Arg?Leu?Arg?Ala?Asp
110 115 120
GGC?GGC?GGG?AAC?GCG?TTC?CCG?TGG?AGC?AAT?GCG?ATG?CTG?CAG?TGG?619
Gly?Gly?Gly?Asn?Ala?Phe?Pro?Trp?Ser?Asn?Ala?Met?Leu?Gln?Trp
125 130 135
CAG?CGC?ACG?GGA?TTC?CAC?TTC?CAG?CCG?CAG?AGG?AAC?TGG?ATG?AAC?664
Gln?Arg?Thr?Gly?Phe?His?Phe?Gln?Pro?Gln?Arg?Asn?Trp?Met?Asn
140 145 150
GAC?CCC?AAT?GGC?CCG?GTG?TAC?TAC?AAG?GGC?TGG?TAC?CAC?CTG?TTC?709
Asp?Pro?Asn?Gly?Pro?Val?Tyr?Tyr?Lys?Gly?Trp?Tyr?His?Leu?Phe
155 160 165
TAC?CAA?TAC?AAC?CCG?GAC?GGC?GCC?ATC?TGG?GGC?AAC?AAG?ATC?GCG?754
Tyr?Gln?Tyr?Asn?Pro?Asp?Gly?Ala?Ile?Trp?Gly?Asn?Lys?Ile?Ala
170 175 180
TGG?GGC?CAC?GCC?GTC?TCC?CGC?GAC?CTC?ATC?CAC?TGG?CGC?CAC?CTC?799
Trp?Gly?His?Ala?Val?Ser?Arg?Asp?Leu?Ile?His?Trp?Arg?His?Leu
185 190 195
CCG?CTG?GCC?ATG?CTG?CCC?GAC?CAG?TGG?TAC?GAC?ACC?AAC?GGC?GTC?844
Pro?Leu?Ala?Met?Leu?Pro?Asp?Gln?Trp?Tyr?Asp?Thr?Asn?Gly?Val
200 205 210
TGG?ACG?GGC?TCC?GCC?ACC?ACG?CTC?CCC?GAC?GGC?CGC?CTC?GCC?ATG?889
Trp?Thr?Gly?Ser?Ala?Thr?Thr?Leu?Pro?Asp?Gly?Arg?Leu?Ala?Met
215 220 225
CTC?TAC?ACC?GGC?TCC?ACC?AAC?ACC?TCC?GTG?CAG?GTG?CAG?TGC?CTC?934
Leu?Tyr?Thr?Gly?Ser?Thr?Asn?Thr?Ser?Val?Gln?Val?Gln?Cys?Leu
230 235 240
GCC?GTC?CCC?GCC?GAC?GAC?GAC?GAC?CCG?CTG?CTC?ACC?AAC?TGG?ACC?979
Ala?Val?Pro?Ala?Asp?Asp?Asp?Asp?Pro?Leu?Leu?Thr?Asn?Trp?Thr
245 250 255
AAG?TAC?GAG?GGC?AAC?CCG?GCG?CTG?TAC?CCG?CCG?CCG?GGG?ATC?GGG?1024
Lys?Tyr?Glu?Gly?Asn?Pro?Ala?Leu?Tyr?Pro?Pro?Pro?Gly?Ile?Gly
260 265 270
CCC?AGG?GAC?TTC?CGC?GAC?CCC?ACC?ACG?GCG?TGG?TTC?GAC?CCG?TCG?1069
Pro?Arg?Asp?Phe?Arg?Asp?Pro?Thr?Thr?Ala?Trp?Phe?Asp?Pro?Ser
275 280 285
GAC?TCC?ACC?TGG?CGC?ATC?GTC?ATC?GGC?TCC?AAG?GAC?GAC?GCC?GAG?1114
Asp?Ser?Thr?Trp?Arg?Ile?Val?Ile?Gly?Ser?Lys?Asp?Asp?Ala?Glu
290 295 300
GGC?GAC?CAC?GCC?GGC?ATC?GCC?GTG?GTG?TAC?CGC?ACC?AGG?GAC?TTC?1159
Gly?Asp?His?Ala?Gly?Ile?Ala?Val?Val?Tyr?Arg?Thr?Arg?Asp?Phe
305 310 315
GTG?CAC?TTC?GAG?CTC?CTC?CCG?GAC?CTG?CTC?CAC?CGC?GTC?GCG?GGG?1204
Val?His?Phe?Glu?Leu?Leu?Pro?Asp?Leu?Leu?His?Arg?Val?Ala?Gly
320 325 330
ACG?GGG?ATG?TGG?GAG?TGC?ATC?GAC?TTC?TAC?CCC?GTC?GCC?ACC?CGC?1249
Thr?Gly?Met?Trp?Glu?Cys?Ile?Asp?Phe?Tyr?Pro?Val?Ala?Thr?Arg
335 340 345
GGC?AAG?GCG?TCC?GGG?AAC?GGC?GTC?GAC?ATG?TCC?GAC?GCC?CTC?GCC?1294
Gly?Lys?Ala?Ser?Gly?Asn?Gly?Val?Asp?Met?Ser?Asp?Ala?Leu?Ala
350 355 360
AAG?AAC?GGC?GCC?GTC?GTC?GGG?GAC?GTG?GTG?CAC?GTC?ATG?AAG?GCC?1339
Lys?Asn?Gly?Ala?Val?Val?Gly?Asp?Val?Val?His?Val?Met?Lys?Ala
365 370 375
AGC?ATG?GAC?GAC?GAC?CGA?CAT?GAC?TAC?TAC?GCG?CTC?GGG?AGG?TAT?1384
Ser?Met?Asp?Asp?Asp?Arg?His?Asp?Tyr?Tyr?Ala?Leu?Gly?Arg?Tyr
380 385 390
GAC?GCG?GCT?GCC?AAC?GCG?TGG?ACG?CCG?CTC?GAC?GCC?GAG?AAG?GAC?1429
Asp?Ala?Ala?Ala?Asn?Ala?Trp?Thr?Pro?Leu?Asp?Ala?Glu?Lys?Asp
395 400 405
GTC?GGC?ACC?GGC?CTG?CGG?TAC?GAC?TGG?GGC?AAG?TTC?TAC?GCG?TCC?1474
Val?Gly?Thr?Gly?Leu?Arg?Tyr?Asp?Trp?Gly?Lys?Phe?Tyr?Ala?Ser
410 415 420
AAG?ACG?TTC?TAC?GAC?CCG?GCC?AAG?CGC?CGC?CGC?GTG?CTC?TGG?GGA?1519
Lys?Thr?Phe?Tyr?Asp?Pro?Ala?Lys?Arg?Arg?Arg?Val?Leu?Trp?Gly
425 430 435
TGG?GTC?GGC?GAG?ACC?GAC?TCG?GAG?CGC?GCT?GAC?GTC?TCC?AAG?GGA?1564
Trp?Val?Gly?Glu?Thr?Asp?Ser?Glu?Arg?Ala?Asp?Val?Ser?Lys?Gly
440 445 450
TGG?GCA?TCG?CTG?CAG?GGT?ATC?CCC?CGG?ACG?GTG?CTG?CTG?GAC?ACC?1609
Trp?Ala?Ser?Leu?Gln?Gly?Ile?Pro?Arg?Thr?Val?Leu?Leu?Asp?Thr
455 460 465
AAG?ACG?GGC?AGC?AAC?CTG?CTG?CAG?TGG?CCC?GTG?GAG?GAA?GTG?GAG?1654
Lys?Thr?Gly?Ser?Asn?Leu?Leu?Gln?Trp?Pro?Val?Glu?Glu?Val?Glu
470 475 480
ACG?CTG?CGC?ACC?AAC?TCC?ACG?GAC?CTC?AGC?GGC?ATC?ACC?ATC?GAC?1699
Thr?Leu?Arg?Thr?Asn?Ser?Thr?Asp?Leu?Ser?Gly?Ile?Thr?Ile?Asp
485 490 495
TAC?GGC?TCC?ACG?TTC?CCG?CTC?AAC?CTC?CGC?CGC?GCC?ACG?CAG?CTG?1744
Tyr?Gly?Ser?Thr?Phe?Pro?Leu?Asn?Leu?Arg?Arg?Ala?Thr?Gln?Leu
500 505 510
GAC?ATC?GAG?GCG?GAG?TTC?GAG?CTG?GAC?CGC?CGC?GCC?GTC?ATG?TCC?1789
Asp?Ile?Glu?Ala?Glu?Phe?Glu?Leu?Asp?Arg?Arg?Ala?Val?Met?Ser
515 520 525
CTC?AAC?GAG?GCC?GAC?GTG?GGC?TAC?AAC?TGC?AGC?ACC?AGC?GGC?GGC?1834
Leu?Asn?Glu?Ala?Asp?Val?Gly?Tyr?Asn?Cys?Ser?Thr?Ser?Gly?Gly
530 535 540
GCC?GCC?GCC?CGC?GGC?GCG?CTG?GGC?CCC?TTC?GGC?CTG?CTC?GTC?CTC 1879
Ala?Ala?Ala?Arg?Gly?Ala?Leu?Gly?Pro?Phe?Gly?Leu?Leu?Val?Leu
545 550 555
ACC?GAC?AAG?CAC?CTG?CAC?GAG?CAG?ACG?GCC?GTC?TAC?TTC?TAC?GTG?1924
Thr?Asp?Lys?His?Leu?His?Glu?Gln?Thr?Ala?Val?Tyr?Phe?Tyr?Val
560 565 570
GCC?AAA?GGC?CTG?GAC?GGC?TCC?CTC?ACC?ACG?CAC?TTC?TGC?CAG?GAC?1969
Ala?Lys?Gly?Leu?Asp?Gly?Ser?Leu?Thr?Thr?His?Phe?Cys?Gln?Asp
575 580 585
GAG?TCC?CGG?TCG?TCC?AGC?GCC?AAC?GAC?ATC?GTC?AAG?CGC?GTC?GTC?2014
Glu?Ser?Arg?Ser?Ser?Ser?Ala?Asn?Asp?Ile?Val?Lys?Arg?Val?Val
590 595 600
GGC?AGC?GCC?GTC?CCC?GTG?CTG?GAG?GAC?GAG?ACC?ACA?CTC?TCG?CTT?2059
Gly?Ser?Ala?Val?Pro?Val?Leu?Glu?Asp?Glu?Thr?Thr?Leu?Ser?Leu
605 610 615
CGC?GTG?CTC?GTC?GAC?CAC?TCC?ATC?GTC?GAG?AGC?TTC?GCG?CAG?GGT?2104
Arg?Val?Leu?Val?Asp?His?Ser?Ile?Val?Glu?Ser?Phe?Ala?Gln?Gly
620 625 630
GGA?AGG?TCA?ACG?GCC?ACC?TCG?CGC?GTC?TAC?CCC?ACC?AAG?GCC?ATC?2149
Gly?Arg?Ser?Thr?Ala?Thr?Ser?Arg?Val?Tyr?Pro?Thr?Lys?Ala?Ile
635 640 645
TAC?GCC?AAC?GCC?GGC?GTG?TTC?CTC?TTC?AAC?AAC?GCC?ACC?GCC?GCG?2194
Tyr?Ala?Asn?Ala?Gly?Val?Phe?Leu?Phe?Asn?Asn?Ala?Thr?Ala?Ala
650 655 660
CGC?GTC?ACC?GCC?AAG?AAG?CTC?GTC?GTC?CAC?GAG?ATG?GAC?TCG?TCC?2239
Arg?Val?Thr?Ala?Lys?Lys?Leu?Val?Val?His?Glu?Met?Asp?Ser?Ser
665 670 675
TAC?AAC?CAC?GAC?TAC?ATG?GTC?ACG?GAC?ATC?2269
Tyr?Asn?His?Asp?Tyr?Met?Val?Thr?Asp?Ile
680 685
TGATGCTGCT GCTGCTGCTG CTGCTGCTGA CCCGTCGTCC ATCCAACCCA CCGCTGCACC?2329
CAATTTTTTG AACCCATATA TAGCGAAGCA TCTTCTTGTA CCTAAAAAAA AAAAAAAA ?2387
<210>2
<211>685
<212>PRT
< 213>sugarcane (Saccharum officinarum)
<400>2
Met?Glu?Thr?Arg?Asp?Thr?Thr?Ala?Pro?Leu?Pro?Tyr?Ser?Tyr?Thr
5 10 15
Pro?Leu?Pro?Ala?Ala?Asp?Ala?Ala?Ser?Ala?Glu?Val?Thr?Gly?Thr
20 25 30
Gly?Gly?Arg?Ser?Arg?Arg?Arg?Ser?Leu?Cys?Ala?Ala?Ala?Leu?Val
35 40 45
Leu?Ser?Ala?Ala?Leu?Leu?Leu?Ala?Val?Ala?Ala?Leu?Ala?Ala?Ala
50 55 60
Gly?Arg?Arg?Pro?Thr?Thr?Ala?Val?Gly?Glu?Thr?Ala?Gly?Val?Gly
65 70 75
Val?Val?Pro?Gly?Val?Gly?Thr?Pro?Gln?Ala?Thr?Ser?Thr?Arg?Ser
80 85 90
Ile?Ser?Arg?Gly?Pro?Asp?Ala?Gly?Val?Ser?Glu?Lys?Thr?Ser?Gly
95 100 105
Ala?Trp?Ser?Gly?Val?Val?Asp?Asp?Gly?Gly?Arg?Leu?Arg?Ala?Asp
110 115 120
Gly?Gly?Gly?Asn?Ala?Phe?Pro?Trp?Ser?Asn?Ala?Met?Leu?Gln?Trp
125 130 135
Gln?Arg?Thr?Gly?Phe?His?Phe?Gln?Pro?Gln?Arg?Asn?Trp?Met?Asn
140 145 150
Asp?Pro?Asn?Gly?Pro?Val?Tyr?Tyr?Lys?Gly?Trp?Tyr?His?Leu?Phe
155 160 165
Tyr?Gln?Tyr?Asn?Pro?Asp?Gly?Ala?Ile?Trp?Gly?Asn?Lys?Ile?Ala
170 175 180
Trp?Gly?His?Ala?Val?Ser?Arg?Asp?Leu?Ile?His?Trp?Arg?His?Leu
185 190 195
Pro?Leu?Ala?Met?Leu?Pro?Asp?Gln?Trp?Tyr?Asp?Thr?Asn?Gly?Val
200 205 210
Trp?Thr?Gly?Ser?Ala?Thr?Thr?Leu?Pro?Asp?Gly?Arg?Leu?Ala?Met
215 220 225
Leu?Tyr?Thr?Gly?Ser?Thr?Asn?Thr?Ser?Val?Gln?Val?Gln?Cys?Leu
230 235 240
Ala?Val?Pro?Ala?Asp?Asp?Asp?Asp?Pro?Leu?Leu?Thr?Asn?Trp?Thr
245 250 255
Lys?Tyr?Glu?Gly?Asn?Pro?Ala?Leu?Tyr?Pro?Pro?Pro?Gly?Ile?Gly
260 265 270
Pro?Arg?Asp?Phe?Arg?Asp?Pro?Thr?Thr?Ala?Trp?Phe?Asp?Pro?Ser
275 280 285
Asp?Ser?Thr?Trp?Arg?Ile?Val?Ile?Gly?Ser?Lys?Asp?Asp?Ala?Glu
290 295 300
Gly?Asp?His?Ala?Gly?Ile?Ala?Val?Val?Tyr?Arg?Thr?Arg?Asp?Phe
305 310 315
Val?His?Phe?Glu?Leu?Leu?Pro?Asp?Leu?Leu?His?Arg?Val?Ala?Gly
320 325 330
Thr?Gly?Met?Trp?Glu?Cys?Ile?Asp?Phe?Tyr?Pro?Val?Ala?Thr?Arg
335 340 345
Gly?Lys?Ala?Ser?Gly?Asn?Gly?Val?Asp?Met?Ser?Asp?Ala?Leu?Ala
350 355 360
Lys?Asn?Gly?Ala?Val?Val?Gly?Asp?Val?Val?His?Val?Met?Lys?Ala
365 370 375
Ser?Met?Asp?Asp?Asp?Arg?His?Asp?Tyr?Tyr?Ala?Leu?Gly?Arg?Tyr
380 385 390
Asp?Ala?Ala?Ala?Asn?Ala?Trp?Thr?Pro?Leu?Asp?Ala?Glu?Lys?Asp
395 400 405
Val?Gly?Thr?Gly?Leu?Arg?Tyr?Asp?Trp?Gly?Lys?Phe?Tyr?Ala?Ser
410 415 420
Lys?Thr?Phe?Tyr?Asp?Pro?Ala?Lys?Arg?Arg?Arg?Val?Leu?Trp?Gly
425 430 435
Trp?Val?Gly?Glu?Thr?Asp?Ser?Glu?Arg?Ala?Asp?Val?Ser?Lys?Gly
440 445 450
Trp?Ala?Ser?Leu?Gln?Gly?Ile?Pro?Arg?Thr?Val?Leu?Leu?Asp?Thr
455 460 465
Lys?Thr?Gly?Ser?Asn?Leu?Leu?Gln?Trp?Pro?Val?Glu?Glu?Val?Glu
470 475 480
Thr?Leu?Arg?Thr?Asn?Ser?Thr?Asp?Leu?Ser?Gly?Ile?Thr?Ile?Asp
485 490 495
Tyr?Gly?Ser?Thr?Phe?Pro?Leu?Asn?Leu?Arg?Arg?Ala?Thr?Gln?Leu
500 505 510
Asp?Ile?Glu?Ala?Glu?Phe?Glu?Leu?Asp?Arg?Arg?Ala?Val?Met?Ser
515 520 525
Leu?Asn?Glu?Ala?Asp?Val?Gly?Tyr?Asn?Cys?Ser?Thr?Ser?Gly?Gly
530 535 540
Ala?Ala?Ala?Arg?Gly?Ala?Leu?Gly?Pro?Phe?Gly?Leu?Leu?Val?Leu
545 550 555
Thr?Asp?Lys?His?Leu?His?Glu?Gln?Thr?Ala?Val?Tyr?Phe?Tyr?Val
560 565 570
Ala?Lys?Gly?Leu?Asp?Gly?Ser?Leu?Thr?Thr?His?Phe?Cys?Gln?Asp
575 580 585
Glu?Ser?Arg?Ser?Ser?Ser?Ala?Asn?Asp?Ile?Val?Lys?Arg?Val?Val
590 595 600
Gly?Ser?Ala?Val?Pro?Val?Leu?Glu?Asp?Glu?Thr?Thr?Leu?Ser?Leu
605 610 615
Arg?Val?Leu?Val?Asp?His?Ser?Ile?Val?Glu?Ser?Phe?Ala?Gln?Gly
620 625 630
Gly?Arg?Ser?Thr?Ala?Thr?Ser?Arg?Val?Tyr?Pro?Thr?Lys?Ala?Ile
635 640 645
Tyr?Ala?Asn?Ala?Gly?Val?Phe?Leu?Phe?Asn?Asn?Ala?Thr?Ala?Ala
650 655 660
Arg?Val?Thr?Ala?Lys?Lys?Leu?Val?Val?His?Glu?Met?Asp?Ser?Ser
665 670 675
Tyr?Asn?His?Asp?Tyr?Met?Val?Thr?Asp?Ile
680 685
Sequence table
Sequence table 2
< 110>Guangxi University
< 120>sugarcane soluble acid invertase gene (SoINV2) gene and protein sequence
<160>2
<210>l
<211>2429
<212>DNA
< 213>sugarcane (Saccharum officinarum)
<220>
<22l>CDS
<222>(227)...(2314)
<220>
<22l>5’UTP
<222>(1)...(226)
<220>
<22l>3’UTP
<222>(2315)...(2429)
<400>l
GAAACCACAT CGCTCGTCCA ATGTGTTGTT GAAATCTAGT TACAATCGAG?TCGAGTCGCC ?60
GCATTGCTCG CTTCCGCGGC TCCAGATCGA ATCGGCCGGC GAGGAGTCGG?TCATCGTCGC 120
TCCGGCCGCC GCCGTAGCCG CCCCTAGAGA GAGTCGCCCG CCGTCGTAAC CGTAACACAA?180
GTCGCCCGGC GGCCTCCTTC CGATCGATCC TCTTCCGTCG TCGGCA ?226
ATG?GAG?ACC?CGG?GAC?ACG?ACG?GCG?CCG?CTC?CCC?TAC?TCG?TAC?ACG?271
Met?Glu?Thr?Arg?Asp?Thr?Thr?Ala?Pro?Leu?Pro?Tyr?Ser?Tyr?Thr
5 10 15
CCG?CTG?CCG?GCC?GCC?GAC?GCC?GCG?TCC?GCC?GAG?GTC?ACC?GGC?ACC?316
Pro?Leu?Pro?Ala?Ala?Asp?Ala?Ala?Ser?Ala?Glu?Val?Thr?Gly?Thr
20 25 30
GGC?CAC?CGC?GGC?GGC?GGC?AGG?AGC?AGG?CGT?AGT?TCC?CTC?TGC?GCC?361
Gly?His?Arg?Gly?Gly?Gly?Arg?Ser?Arg?Arg?Ser?Ser?Leu?Cys?Ala
35 40 45
GCG?GCG?CTC?GTC?CTC?TCC?GCC?GCG?CTG?CTC?CTC?GCC?GTG?GCC?GCG?406
Ala?Ala?Leu?Val?Leu?Ser?Ala?Ala?Leu?Leu?Leu?Ala?Val?Ala?Ala
50 55 60
CTC?GCC?GGC?GTC?GGC?GGC?CGC?GTC?GCC?GTC?GTC?CCC?CGC?CCA?ACG?451
Leu?Ala?Gly?Val?Gly?Gly?Arg?Val?Ala?Val?Val?Pro?Arg?Pro?Thr
65 70 75
ACC?GCG?GTG?GGA?GAA?ACG?GCC?GGC?GTC?GGC?GTC?GGC?CCT?GGC?GCG?496
Thr?Ala?Val?Gly?Glu?Thr?Ala?Gly?Val?Gly?Val?Gly?Pro?Gly?Ala
80 85 90
GGG?ACA?CCA?CAG?GCG?ACG?TCG?ACC?AGG?AGC?ATC?AGC?AGG?GGC?CCC?541
Gly?Thr?Pro?Gln?Ala?Thr?Ser?Thr?Arg?Ser?lle?Ser?Arg?Gly?Pro
95 100 105
GAC?GCC?GGC?GTG?TCG?GAG?AAG?ACG?TCC?GGC?GCG?TGG?AGC?GGC?GTC?586
Asp?Ala?Gly?Val?Ser?Glu?Lys?Thr?Ser?Gly?Ala?Trp?Ser?Gly?Val
110 115 120
GTC?GAC?GAT?GGC?GGG?AGG?CTC?CGT?GCT?GAC?GGC?GGC?GGG?AAC?GCG?631
Val?Asp?Asp?Gly?Gly?Arg?Leu?Arg?Ala?Asp?Gly?Gly?Gly?Asn?Ala
125 130 135
TTC?CCG?TGG?AGC?AAT?GCG?ATG?CTG?CAG?TGG?CAG?CGC?ACG?GGA?TTC?676
Phe?Pro?Trp?Ser?Asn?Ala?Met?Leu?Gln?Trp?Gln?Arg?Thr?Gly?Phe
140 145 150
CAC?TTC?CAG?CCG?CAG?AGG?AAC?TGG?ATG?AAC?GAC?CCC?AAT?GGC?CCG?721
His?Phe?Gln?Pro?Gln?Arg?Asn?Trp?Met?Asn?Asp?Pro?Asn?Gly?Pro
155 160 165
GTG?TAC?TAC?AAG?GGC?TGG?TAC?CAC?CTG?TTC?TAC?CAA?TAC?AAC?CCG?766
Val?Tyr?Tyr?Lys?Gly?Trp?Tyr?His?Leu?Phe?Tyr?Gln?Tyr?Asn?Pro
170 175 180
GAC?GGC?GCC?ATC?TGG?GGC?AAC?AAG?ATC?GCG?TGG?GGC?CAC?GCC?GTC?811
Asp?Gly?Ala?Ile?Trp?Gly?Asn?Lys?Ile?Ala?Trp?Gly?His?Ala?Val
185 190 195
TCC?CGC?GAC?CTC?ATC?CAC?TGG?CGC?CAC?CTC?CCG?CTG?GCC?ATG?CTG?856
Ser?Arg?Asp?Leu?Ile?His?Trp?Arg?His?Leu?Pro?Leu?Ala?Met?Leu
200 205 210
CCC?GAC?CAG?TGG?TAC?GAC?ACC?AAC?GGC?GTC?TGG?ACG?GGC?TCC?GCC?901
Pro?Asp?Gln?Trp?Tyr?Asp?Thr?Asn?Gly?Val?Trp?Thr?Gly?Ser?Ala
215 220 225
ACC?ACG?CTC?CCC?GAC?GGC?CGC?CTC?GCC?ATG?CTC?TAC?ACC?GGC?TCC?946
Thr?Thr?Leu?Pro?Asp?Gly?Arg?Leu?Ala?Met?Leu?Tyr?Thr?Gly?Ser
230 235 240
ACC?AAC?ACC?TCC?GTG?CAG?GTG?CAG?TGC?CTC?GCC?GTC?CCC?GCC?GGC?991
Thr?Asn?Thr?Ser?Val?Gln?Val?Gln?Cys?Leu?Ala?Val?Pro?Ala?Asp
245 250 255
GAC?GAC?GAC?CCG?CTG?CTC?ACC?AAC?TGG?ACC?AAG?TAC?GAG?GGC?AAC?1036
Asp?Asp?Asp?Pro?Leu?Leu?Thr?Asn?Trp?Thr?Lys?Tyr?Glu?Gly?Asn
260 265 270
CCG?GCG?CTG?TAC?CCG?CCG?CCG?GGG?ATC?GGG?CCC?AGG?GAC?TTC?CGC?1081
Pro?Ala?Leu?Tyr?Pro?Pro?Pro?Gly?Ile?Gly?Pro?Arg?Asp?Phe?Arg
275 280 285
GAC?CCC?ACC?ACG?GCG?TGG?TTC?GAC?CCG?TCG?GAC?TCC?ACC?TGG?CGC?1126
Asp?Pro?Thr?Thr?Ala?Trp?Phe?Asp?Pro?Ser?Asp?Ser?Thr?Trp?Arg
290 295 300
ATC?GTC?ATC?GGC?TCC?AAG?GAC?GAC?GCC?GAG?GGC?GAC?CAC?GCC?GGC?1171
Ile?Val?Ile?Gly?Ser?Lys?Asp?Asp?Ala?Glu?Gly?Asp?His?Ala?Gly
305 310 315
ATC?GCC?GTG?GTG?TAC?CGC?ACC?AGG?GAC?TTC?GTG?CAC?TTC?GAG?CTC?1216
Ile?Ala?Val?Val?Tyr?Arg?Thr?Arg?Asp?Phe?Val?His?Phe?Glu?Leu
320 325 330
CTC?CCG?GAC?CTG?CTC?CAC?CGC?GTC?GCG?GGG?ACG?GGG?ATG?TGG?GAG?1261
Leu?Pro?Asp?Leu?Leu?His?Arg?Val?Ala?Gly?Thr?Gly?Met?Trp?Glu
335 340 345
TGC?ATC?GAC?TTC?TAC?CCC?GTC?GCC?ACC?CGC?GGC?AAG?GCG?TCC?GGG?1306
Cys?Ile?Asp?Phe?Tyr?Pro?Val?Ala?Thr?Arg?Gly?Lys?Ala?Ser?Gly
350 355 360
AAC?GGC?GTC?GAC?ATG?TCC?GAC?GCC?CTC?GCC?AAG?AAC?GGC?GCC?GTC?1351
Asn?Gly?Val?Asp?Met?Ser?Asp?Ala?Leu?Ala?Lys?Asn?Gly?Ala?Val
365 370 375
GTC?GGG?GAC?GTG?GTG?CAC?GTC?ATG?AAG?GCC?AGC?ATG?GAC?GAC?GAC?1396
Val?Gly?Asp?Val?Val?His?Val?Met?Lys?Ala?Ser?Met?Asp?Asp?Asp
380 385 390
CGA?CAT?GAC?TAC?TAC?GCG?CTC?GGG?AGG?TAT?GAC?GCG?GCT?GCC?AAC?1441
Arg?His?Asp?Tyr?Tyr?Ala?Leu?Gly?Arg?Tyr?Asp?Ala?Ala?Ala?Asn
395 400 405
GCG?TGG?ACG?CCG?CTC?GAC?GCC?GAG?AAG?GAC?GTC?GGC?ACC?GGC?CTG?1486
Ala?Trp?Thr?Pro?Leu?Asp?Ala?Glu?Lys?Asp?Val?Gly?Thr?Gly?Leu
410 415 420
CGG?TAC?GAC?TGG?GGC?AAG?TTC?TAC?GCG?TCC?AAG?ACG?TTC?TAC?GAC?1531
Arg?Tyr?Asp?Trp?Gly?Lys?Phe?Tyr?Ala?Ser?Lys?Thr?Phe?Tyr?Asp
425 430 435
CCG?GCC?AAG?CGC?CGC?CGC?GTG?CTC?TGG?GGA?TGG?GTC?GGC?GAG?ACC?1576
Pro?Ala?Lys?Arg?Arg?Arg?Val?Leu?Trp?Gly?Trp?Val?Gly?Glu?Thr
440 445 450
GAC?TCG?GAG?CGC?GCT?GAC?GTC?TCC?AAG?GGA?TGG?GCA?TCG?CTG?CAG?1621
Asp?Ser?Glu?Arg?Ala?Asp?Val?Ser?Lys?Gly?Trp?Ala?Ser?Leu?Gln
455 460 465
GGT?ATC?CCC?CGG?ACG?GTG?CTG?CTG?GAC?ACC?AAG?ACG?GGC?AGC?AAC?1666
Gly?Ile?Pro?Arg?Thr?Val?Leu?Leu?Asp?Thr?Lys?Thr?Gly?Ser?Asn
470 475 480
CTG?CTG?CAG?TGG?CCC?GTG?GAG?GAA?GTG?GAG?ACG?CTG?CGC?ACC?AAC?1711
Leu?Leu?Gln?Trp?Pro?Val?Glu?Glu?Val?Glu?Thr?Leu?Arg?Thr?Asn
485 490 495
TCC?ACG?GAC?CTC?AGC?GGC?ATC?ACC?ATC?GAC?TAC?GGC?TCC?ACG?TTC?1756
Ser?Thr?Asp?Leu?Ser?Gly?Ile?Thr?Ile?Asp?Tyr?Gly?Ser?Thr?Phe
500 505 510
CCG?CTC?AAC?CTC?CGC?CGC?GCC?ACG?CAG?CTG?GAC?ATC?GAG?GCG?GAG?1801
Pro?Leu?Asn?Leu?Arg?Arg?Ala?Thr?Gln?Leu?Asp?Ile?Glu?Ala?Glu
515 520 525
TTC?GAG?CTG?GAC?CGC?CGC?GCC?GTC?ATG?TCC?CTC?AAC?GAG?GCC?GAC?1846
Phe?Glu?Leu?Asp?Arg?Arg?Ala?Val?Met?Ser?Leu?Asn?Glu?Ala?Asp
530 535 540
GTG?GGC?TAC?AAC?TGC?AGC?ACC?AGC?GGC?GGC?GCC?GCC?GCC?CGC?GGC?1891
Val?Gly?Tyr?Asn?Cys?Ser?Thr?Ser?Gly?Gly?Ala?Ala?Ala?Arg?Gly
545 550 555
GCG?CTG?GGC?CCC?TTC?GGC?CTG?CTC?GTC?CTC?ACC?GAC?AAG?CAC?CTG?1936
Ala?Leu?Gly?Pro?Phe?Gly?Leu?Leu?Val?Leu?Thr?Asp?Lys?His?Leu
560 565 570
CAC?GAG?CAG?ACG?GCC?GTC?TAC?TTC?TAC?GTG?GCC?AAA?GGC?CTG?GAC?1981
His?Glu?Gln?Thr?Ala?Val?Tyr?Phe?Tyr?Val?Ala?Lys?Gly?Leu?Asp
575 580 585
GGC?TCC?CTC?ACC?ACG?CAC?TTC?TGC?CAG?GAC?GAG?TCC?CGG?TCG?TCC?2026
Gly?Ser?Leu?Thr?Thr?His?Phe?Cys?Gln?Asp?Glu?Ser?Arg?Ser?Ser
590 595 600
AGC?GCC?AAC?GAC?ATC?GTC?AAG?CGC?GTC?GTC?GGC?AGC?GCC?GTC?CCC?2071
Ser?Ala?Asn?Asp?Ile?Val?Lys?Arg?Val?Val?Gly?Ser?Ala?Val?Pro
605 610 615
GTG?CTG?GAG?GAC?GAG?ACC?ACA?CTC?TCG?CTT?CGC?GTG?CTC?GTC?GAC?2116
Val?Leu?Glu?Asp?Glu?Thr?Thr?Leu?Ser?Leu?Arg?Val?Leu?Val?Asp
620 625 630
CAC?TCC?ATC?GTC?GAG?AGC?TTC?GCG?CAG?GGT?GGA?AGG?TCA?ACG?GCC?2161
His?Ser?Ile?Val?Glu?Ser?Phe?Ala?Gln?Gly?Gly?Arg?Ser?Thr?Ala
635 640 645
ACC?TCG?CGC?GTC?TAC?CCC?ACC?AAG?GCC?ATC?TAC?GCC?AAC?GCC?GGC?2206
Thr?Ser?Arg?Val?Tyr?Pro?Thr?Lys?Ala?Ile?Tyr?Ala?Asn?Ala?Gly
650 655 660
GTG?TTC?CTC?TTC?AAC?AAC?GCC?ACC?GCC?GCG?CGC?GTC?ACC?GCC?AAG?2251
Val?Phe?Leu?Phe?Asn?Asn?Ala?Thr?Ala?Ala?Arg?Val?Thr?Ala?Lys
665 670 675
AAG?CTC?GTC?GTC?CAC?GAG?ATG?GAC?TCG?TCC?TAC?AAC?CAC?GAC?TAC?2296
Lys?Leu?Val?Val?His?Glu?Met?Asp?Ser?Ser?Tyr?Asn?His?Asp?Tyr
680 685 690
ATG?GTC?ACG?GAC?ATC?2311
Met?Val?Thr?Asp?Ile
695
TGATGCTGCT GCTGCTGCTG CTGCTGCTGA CCCGTCGTCC ATCCAACCCA CCGCTGCACC?2371
CAATTTTTTG AACCCATATA TAGCGAAGCA TCTTCTTGTA CCTAAAAAAA AAAAAAAA ?2429
<210>2
<211>695
<212>PRT
< 213>sugarcane (Saccharum officinarum)
<400>2
Met?Glu?Thr?Arg?Asp?Thr?Thr?Ala?Pro?Leu?Pro?Tyr?Ser?Tyr?Thr
5 10 15
Pro?Leu?Pro?Ala?Ala?Asp?Ala?Ala?Ser?Ala?Glu?Val?Thr?Gly?Thr
20 25 30
Gly?His?Arg?Gly?Gly?Gly?Arg?Ser?Arg?Arg?Ser?Ser?Leu?Cys?Ala
35 40 45
Ala?Ala?Leu?Val?Leu?Ser?Ala?Ala?Leu?Leu?Leu?Ala?Val?Ala?Ala
50 55 60
Leu?Ala?Gly?Val?Gly?Gly?Arg?Val?Ala?Val?Val?Pro?Arg?Pro?Thr
65 70 75
Thr?Ala?Val?Gly?Glu?Thr?Ala?Gly?Val?Gly?Val?Gly?Pro?Gly?Ala
80 85 ?90
Gly?Thr?Pro?Gln?Ala?Thr?Ser?Thr?Arg?Ser?Ile?Ser?Arg?Gly?Pro
95 100 105
Asp?Ala?Gly?Val?Ser?Glu?Lys?Thr?Ser?Gly?Ala?Trp?Ser?Gly?Val
110 115 120
Val?Asp?Asp?Gly?Gly?Arg?Leu?Arg?Ala?Asp?Gly?Gly?Gly?Asn?Ala
125 130 135
Phe?Pro?Trp?Ser?Asn?Ala?Met?Leu?Gln?Trp?Gln?Arg?Thr?Gly?Phe
140 145 150
His?Phe?Gln?Pro?Gln?Arg?Asn?Trp?Met?Asn?Asp?Pro?Asn?Gly?Pro
155 160 165
Val?Tyr?Tyr?Lys?Gly?Trp?Tyr?His?Leu?Phe?Tyr?Gln?Tyr?Asn?Pro
170 175 180
Asp?Gly?Ala?Ile?Trp?Gly?Asn?Lys?Ile?Ala?Trp?Gly?His?Ala?Val
185 190 195
Ser?Arg?Asp?Leu?Ile?His?Trp?Arg?His?Leu?Pro?Leu?Ala?Met?Leu
200 205 210
Pro?Asp?Gln?Trp?Tyr?Asp?Thr?Asn?Gly?Val?Trp?Thr?Gly?Ser?Ala
215 220 225
Thr?Thr?Leu?Pro?Asp?Gly?Arg?Leu?Ala?Met?Leu?Tyr?Thr?Gly?Ser
230 235 240
Thr?Asn?Thr?Ser?Val?Gln?Val?Gln?Cys?Leu?Ala?Val?Pro?Ala?Asp
245 250 255
Asp?Asp?Asp?Pro?Leu?Leu?Thr?Asn?Trp?Thr?Lys?Tyr?Glu?Gly?Asn
260 265 270
Pro?Ala?Leu?Tyr?Pro?Pro?Pro?Gly?Ile?Gly?Pro?Arg?Asp?Phe?Arg
275 280 285
Asp?Pro?Thr?Thr?Ala?Trp?Phe?Asp?Pro?Ser?Asp?Ser?Thr?Trp?Arg
290 295 300
Ile?Val?Ile?Gly?Ser?Lys?Asp?Asp?Ala?Glu?Gly?Asp?His?Ala?Gly
305 310 315
Ile?Ala?Val?Val?Tyr?Arg?Thr?Arg?Asp?Phe?Val?His?Phe?Glu?Leu
320 325 330
Leu?Pro?Asp?Leu?Leu?His?Arg?Val?Ala?Gly?Thr?Gly?Met?Trp?Glu
335 340 345
Cys?Ile?Asp?Phe?Tyr?Pro?Val?Ala?Thr?Arg?Gly?Lys?Ala?Ser?Gly
350 355 360
Asn?Gly?Val?Asp?Met?Ser?Asp?Ala?Leu?Ala?Lys?Asn?Gly?Ala?Val
365 370 375
Val?Gly?Asp?Val?Val?His?Val?Met?Lys?Ala?Ser?Met?Asp?Asp?Asp
380 385 390
Arg?His?Asp?Tyr?Tyr?Ala?Leu?Gly?Arg?Tyr?Asp?Ala?Ala?Ala?Asn
395 400 405
Ala?Trp?Thr?Pro?Leu?Asp?Ala?Glu?Lys?Asp?Val?Gly?Thr?Gly?Leu
410 415 420
Arg?Tyr?Asp?Trp?Gly?Lys?Phe?Tyr?Ala?Ser?Lys?Thr?Phe?Tyr?Asp
425 430 435
Pro?Ala?Lys?Arg?Arg?Arg?Val?Leu?Trp?Gly?Trp?Val?Gly?Glu?Thr
440 445 450
Asp?Ser?Glu?Arg?Ala?Asp?Val?Ser?Lys?Gly?Trp?Ala?Ser?Leu?Gln
455 460 465
Gly?Ile?Pro?Arg?Thr?Val?Leu?Leu?Asp?Thr?Lys?Thr?Gly?Ser?Asn
470 475 480
Leu?Leu?Gln?Trp?Pro?Val?Glu?Glu?Val?Glu?Thr?Leu?Arg?Thr?Asn
485 490 495
Ser?Thr?Asp?Leu?Ser?Gly?Ile?Thr?Ile?Asp?Tyr?Gly?Ser?Thr?Phe
500 505 510
Pro?Leu?Asn?Leu?Arg?Arg?Ala?Thr?Gln?Leu?Asp?Ile?Glu?Ala?Glu
515 520 525
Phe?Glu?Leu?Asp?Arg?Arg?Ala?Val?Met?Ser?Leu?Asn?Glu?Ala?Asp
530 535 540
Val?Gly?Tyr?Asn?Cys?Ser?Thr?Ser?Gly?Gly?Ala?Ala?Ala?Arg?Gly
545 550 555
Ala?Leu?Gly?Pro?Phe?Gly?Leu?Leu?Val?Leu?Thr?Asp?Lys?His?Leu
560 565 570
His?Glu?Gln?Thr?Ala?Val?Tyr?Phe?Tyr?Val?Ala?Lys?Gly?Leu?Asp
575 580 585
Gly?Ser?Leu?Thr?Thr?His?Phe?Cys?Gln?Asp?Glu?Ser?Arg?Ser?Ser
590 595 600
Ser?Ala?Asn?Asp?Ile?Val?Lys?Arg?Val?Val?Gly?Ser?Ala?Val?Pro
605 610 615
Val?Leu?Glu?Asp?Glu?Thr?Thr?Leu?Ser?Leu?Arg?Val?Leu?Val?Asp
620 625 630
His?Ser?Ile?Val?Glu?Ser?Phe?Ala?Gln?Gly?Gly?Arg?Ser?Thr?Ala
635 640 645
Thr?Ser?Arg?Val?Tyr?Pro?Thr?Lys?Ala?Ile?Tyr?Ala?Asn?Ala?Gly
650 655 660
Val?Phe?Leu?Phe?Asn?Asn?Ala?Thr?Ala?Ala?Arg?Val?Thr?Ala?Lys
665 670 675
Lys?Leu?Val?Val?His?Glu?Met?Asp?Ser?Ser?Tyr?Asn?His?Asp?Tyr
680 685 690
Met?Val?Thr?Asp?Ile
695
Sequence table
Sequence table 3
< 110>Guangxi University
< 120>sugarcane soluble acid invertase (SoINV3) gene and protein sequence
<160>2
<210>l
<211>2373
<212>DNA
< 213>sugarcane (Saccharum officinarum)
<220>
<22l>CDS
<222>(168)...(2258)
<220>
<22l>5’UTP
<222>(1)...(167)
<220>
<22l>3’UTP
<222>(2259)...(2373)
<400>l
GAAAGTTGTT GTATGTACTA CTAGATTCTA GTTACAATCG AGTCGCATTG CGCGGCTCCA?60
GATCGAATCG GCCAGGAGTC GGTCATCGTC GTCCGCTCCG CTCCGGCCGC CACCGTAACC?120
GTAACACAAG TCGCCGGCGC CCTCCTTCCG ATCCTCTTCC TTCGACA?167
ATG?GAG?ACC?CGG?GAC?ACG?ACG?GCG?CCG?CTC?CCC?TAC?TCG?TAC?ACG?212
Met?Glu?Thr?Arg?Asp?Thr?Thr?Ala?Pro?Leu?Pro?Tyr?Ser?Tyr?Thr
5 10 15
CCG?CTG?CCG?GCC?GCC?GAC?GCC?GCG?TCC?GCC?GAG?GTC?ACC?GGC?ACC?257
Pro?Leu?Pro?Ala?Ala?Asp?Ala?Ala?Ser?Ala?Glu?Val?Thr?Gly?Thr
20 25 ?30
GGC?GGC?AGC?AGG?AGC?AGG?CGG?AGG?CGG?CCG?CTC?TGC?GCC?GCG?GCG?302
Gly?Gly?Ser?Arg?Ser?Arg?Arg?Arg?Arg?Pro?Leu?Cys?Ala?Ala?Ala
35 40 ?45
CTC?GTC?CTC?TCC?GCC?GCG?CTG?CTC?CTC?GCC?GTG?GCC?GCG?CTC?GCC?347
Leu?Val?Leu?Ser?Ala?Ala?Leu?Leu?Leu?Ala?Val?Ala?Ala?Leu?Ala
50 55 ?60
GGC?GTC?GGC?AGC?CGC?GTC?GCC?GCC?GTC?GTC?CCC?CGC?CCA?ACG?ACC?392
Gly?Val?Gly?Ser?Arg?Val?Ala?Ala?Val?Val?Pro?Arg?Pro?Thr?Thr
65 70 ?75
GCG?GTG?GGA?GAA?ACG?GCC?GGC?GTC?GGC?GTC?GGC?GTC?GTC?CCT?GGC?437
Ala?Val?Gly?Glu?Thr?Ala?Gly?Val?Gly?Val?Gly?Val?Val?Pro?Gly
80 85 ?90
GCG?GGG?ACA?CCA?CAG?GCG?ACG?TCG?ACC?AGG?AGC?CGC?AGC?AGG?GGC?482
Ala?Gly?Thr?Pro?Gln?Ala?Thr?Ser?Thr?Arg?Ser?Arg?Ser?Arg?Gly
95 100 105
CCC?GAT?GCC?GGC?GTG?TCG?GAG?AAG?ACG?TCC?GGC?GTG?TGG?ACC?GGC?527
Pro?Asp?Ala?Gly?Val?Ser?Glu?Lys?Thr?Ser?Gly?Val?Trp?Thr?Gly
110 115 120
GTC?ATC?GAC?GAT?GGC?GCC?AGG?CTG?CGG?ACT?GAC?GCC?GGC?GGC?AAC?572
Val?Ile?Asp?Asp?Gly?Ala?Arg?Leu?Arg?Thr?Asp?Ala?Gly?Gly?Asn
125 130 135
GCG?TTC?CCG?TGG?AGC?AAT?GCG?ATG?CTG?CAG?TGG?CAG?CGC?ACG?GGC?617
Ala?Phe?Pro?Trp?Ser?Asn?Ala?Met?Leu?Gln?Trp?Gln?Arg?Thr?Gly
140 145 150
TTC?CAC?TTC?CAG?CCG?CAG?AGG?AAC?TGG?ATG?AAC?GAC?CCC?AAT?GGC?662
Phe?His?Phe?Gln?Pro?Gln?Arg?Asn?Trp?Met?Asn?Asp?Pro?Asn?Gly
155 160 165
CCG?GTG?TAC?TAC?AAG?GGC?TGG?TAC?CAC?CTG?TTC?TAC?CAA?TAC?AAC?707
Pro?Val?Tyr?Tyr?Lys?Gly?Trp?Tyr?His?Leu?Phe?Tyr?Gln?Tyr?Asn
170 175 180
CCG?GAC?GGC?GCC?ATC?TGG?GGC?AAC?AAG?ATC?GCG?TGG?GGC?CAC?GCC?752
Pro?Asp?Gly?Ala?Ile?Trp?Gly?Asn?Lys?Ile?Ala?Trp?Gly?His?Ala
185 190 195
GTC?TCC?CGC?GAC?CTC?ATC?CAC?TGG?CGC?CAC?CTC?CCG?CTG?GCC?ATG?797
Val?Ser?Arg?Asp?Leu?Ile?His?Trp?Arg?His?Leu?Pro?Leu?Ala?Met
200 205 210
CTG?CCC?GAC?CAG?TGG?TAC?GAC?ACC?AAC?GGC?GTC?TGG?ACG?GGC?TCC?842
Leu?Pro?Asp?Gln?Trp?Tyr?Asp?Thr?Asn?Gly?Val?Trp?Thr?Gly?Ser
215 220 225
GCC?ACC?ACG?CTC?CCC?GAC?GGC?CGC?CTC?GCC?ATG?CTC?TAC?ACC?GGC?887
Ala?Thr?Thr?Leu?Pro?Asp?Gly?Arg?Leu?Ala?Met?Leu?Tyr?Thr?Gly
230 235 240
TCC?ACC?AAC?ACC?TCC?GTG?CAG?GTG?CAG?TGC?CTC?GCC?GTC?CCC?GCC?932
Ser?Thr?Asn?Thr?Ser?Val?Gln?Val?Gln?Cys?Leu?Ala?Val?Pro?Ala
245 250 255
GAC?GAC?GAC?GAC?CCG?CTG?CTC?ACC?AAC?TGG?ACC?AAG?TAC?GAG?GGC?977
Asp?Asp?Asp?Asp?Pro?Leu?Leu?Thr?Asn?Trp?Thr?Lys?Tyr?Glu?Gly
260 265 270
AAC?CCG?GCG?CTG?TAC?CCG?CCG?CCG?GGG?ATC?GGG?CCC?AGG?GAC?TTC?1022
Asn?Pro?Ala?Leu?Tyr?Pro?Pro?Pro?Gly?Ile?Gly?Pro?Arg?Asp?Phe
275 280 285
CGC?GAC?CCC?ACC?ACG?GCG?TGG?TTC?GAC?CCG?TCG?GAC?TCC?ACC?TGG?1067
Arg?Asp?Pro?Thr?Thr?Ala?Trp?Phe?Asp?Pro?Ser?Asp?Ser?Thr?Trp
290 295 300
CGC?ATC?GTC?ATC?GGC?TCC?AAG?GAC?GAC?GCC?GAG?GGC?GAC?CAC?GCC?1112
Arg?Ile?Val?Ile?Gly?Ser?Lys?Asp?Asp?Ala?Glu?Gly?Asp?His?Ala
305 310 315
GGC?ATC?GCC?GTG?GTG?TAC?CGC?ACC?AGG?GAC?TTC?GTG?CAC?TTC?GAG?1157
Gly?Ile?Ala?Val?Val?Tyr?Arg?Thr?Arg?Asp?Phe?Val?His?Phe?Glu
320 325 330
CTC?CTC?CCG?GAC?CTG?CTC?CAC?CGC?GTC?GCG?GGG?ACG?GGG?ATG?TGG?1202
Leu?Leu?Pro?Asp?Leu?Leu?His?Arg?Val?Ala?Gly?Thr?Gly?Met?Trp
335 340 345
GAG?TGC?ATC?GAC?TTC?TAC?CCC?GTC?GCC?ACC?CGC?GGC?AAG?GCG?TCC?1247
Glu?Cys?Ile?Asp?Phe?Tyr?Pro?Val?Ala?Thr?Arg?Gly?Lys?Ala?Ser
350 355 360
GGG?AAC?GGC?GTC?GAC?ATG?TCC?GAC?GCC?CTC?GCC?AAG?AAC?GGC?GCC?1292
Gly?Asn?Gly?Val?Asp?Met?Ser?Asp?Ala?Leu?Ala?Lys?Asn?Gly?Ala
365 370 375
GTC?GTC?GGG?GAC?GTG?GTG?CAC?GTC?ATG?AAG?GCC?AGC?ATG?GAC?GAC?1337
Val?Val?Gly?Asp?Val?Val?His?Val?Met?Lys?Ala?Ser?Met?Asp?Asp
380 385 390
GAC?CGA?CAT?GAC?TAC?TAC?GCG?CTC?GGG?AGG?TAT?GAC?GCG?GCT?GCC?1382
Asp?Arg?His?Asp?Tyr?Tyr?Ala?Leu?Gly?Arg?Tyr?Asp?Ala?Ala?Ala
395 400 405
AAC?GCG?TGG?ACG?CCG?CTC?GAC?GCC?GAG?AAG?GAC?GTC?GGC?ACC?GGC?1427
Asn?Ala?Trp?Thr?Pro?Leu?Asp?Ala?Glu?Lys?Asp?Val?Gly?Thr?Gly
410 415 420
CTG?CGG?TAC?GAC?TGG?GGC?AAG?TTC?TAC?GCG?TCC?AAG?ACG?TTC?TAC?1472
Leu?Arg?Tyr?Asp?Trp?Gly?Lys?Phe?Tyr?Ala?Ser?Lys?Thr?Phe?Tyr
425 430 435
GAC?CCG?GCC?AAG?CGC?CGC?CGC?GTG?CTC?TGG?GGA?TGG?GTC?GGC?GAG?1517
Asp?Pro?Ala?Lys?Arg?Arg?Arg?Val?Leu?Trp?Gly?Trp?Val?Gly?Glu
440 445 450
ACC?GAC?TCG?GAG?CGC?GCT?GAC?GTC?TCC?AAG?GGA?TGG?GCA?TCG?CTG?1562
Thr?Asp?Ser?Glu?Arg?Ala?Asp?Val?Ser?Lys?Gly?Trp?Ala?Ser?Leu
455 460 465
CAG?GGT?ATC?CCC?CGG?ACG?GTG?CTG?CTG?GAC?ACC?AAG?ACG?GGC?AGC?1607
Gln?Gly?Ile?Pro?Arg?Thr?Val?Leu?Leu?Asp?Thr?Lys?Thr?Gly?Ser
470 475 480
AAC?CTG?CTG?CAG?TGG?CCC?GTG?GAG?GAA?GTG?GAG?ACG?CTG?CGC?ACC?1652
Asn?Leu?Leu?Gln?Trp?Pro?Val?Glu?Glu?Val?Glu?Thr?Leu?Arg?Thr
485 490 495
AAC?TCC?ACG?GAC?CTC?AGC?GGC?ATC?ACC?ATC?GAC?TAC?GGC?TCC?ACG?1697
Asn?Ser?Thr?Asp?Leu?Ser?Gly?Ile?Thr?Ile?Asp?Tyr?Gly?Ser?Thr
500 505 510
TTC?CCG?CTC?AAC?CTC?CGC?CGC?GCC?ACG?CAG?CTG?GAC?ATC?GAG?GCG?1742
Phe?Pro?Leu?Asn?Leu?Arg?Arg?Ala?Thr?Gln?Leu?Asp?Ile?Glu?Ala
515 520 525
GAG?TTC?GAG?CTG?GAC?CGC?CGC?GCC?GTC?ATG?TCC?CTC?AAC?GAG?GCC?1787
Glu?Phe?Glu?Leu?Asp?Arg?Arg?Ala?Val?Met?Ser?Leu?Asn?Glu?Ala
530 535 540
GAC?GTG?GGC?TAC?AAC?TGC?AGC?ACC?AGC?GGC?GGC?GCC?GCC?GCC?CGC?1832
Asp?Val?Gly?Tyr?Asn?Cys?Ser?Thr?Ser?Gly?Gly?Ala?Ala?Ala?Arg
545 550 555
GGC?GCG?CTG?GGC?CCC?TTC?GGC?CTG?CTC?GTC?CTC?ACC?GAC?AAG?CAC?1877
Gly?Ala?Leu?Gly?Pro?Phe?Gly?Leu?Leu?Val?Leu?Thr?Asp?Lys?His
560 565 570
CTG?CAC?GAG?CAG?ACG?GCC?GTC?TAC?TTC?TAC?GTG?GCC?AAA?GGC?CTG?1922
Leu?His?Glu?Gln?Thr?Ala?Val?Tyr?Phe?Tyr?Val?Ala?Lys?Gly?Leu
575 580 585
GAC?GGC?TCC?CTC?ACC?ACG?CAC?TTC?TGC?CAG?GAC?GAG?TCC?CGG?TCG?1967
Asp?Gly?Ser?Leu?Thr?Thr?His?Phe?Cys?Gln?Asp?Glu?Ser?Arg?Ser
590 595 600
TCC?AGC?GCC?AAC?GAC?ATC?GTC?AAG?CGC?GTC?GTC?GGC?AGC?GCC?GTC?2012
Ser?Ser?Ala?Asn?Asp?Ile?Val?Lys?Arg?Val?Val?Gly?Ser?Ala?Val
605 610 615
CCC?GTG?CTG?GAG?GAC?GAG?ACC?ACA?CTC?TCG?CTT?CGC?GTG?CTC?GTC?2057
Pro?Val?Leu?Glu?Asp?Glu?Thr?Thr?Leu?Ser?Leu?Arg?Val?Leu?Val
620 625 630
GAC?CAC?TCC?ATC?GTC?GAG?AGC?TTC?GCG?CAG?GGT?GGA?AGG?TCA?ACG?2102
Asp?His?Ser?Ile?Val?Glu?Ser?Phe?Ala?Gln?Gly?Gly?Arg?Ser?Thr
635 640 645
GCC?ACC?TCG?CGC?GTC?TAC?CCC?ACC?AAG?GCC?ATC?TAC?GCC?AAC?GCC?2147
Ala?Thr?Ser?Arg?Val?Tyr?Pro?Thr?Lys?Ala?Ile?Tyr?Ala?Asn?Ala
650 655 660
GGC?GTG?TTC?CTC?TTC?AAC?AAC?GCC?ACC?GCC?GCG?CGC?GTC?ACC?GCC?2192
Gly?Val?Phe?Leu?Phe?Asn?Asn?Ala?Thr?Ala?Ala?Arg?Val?Thr?Ala
665 670 675
AAG?AAG?CTC?GTC?GTC?CAC?GAG?ATG?GAC?TCG?TCC?TAC?AAC?CAC?GAC?2237
Lys?Lys?Leu?Val?Val?His?Glu?Met?Asp?Ser?Ser?Tyr?Ash?His?Asp
680 685 690
TAC?ATG?GTC?ACG?GAC?ATC?2255
Tyr?Met?Val?Thr?Asp?Ile
695?696
TGATGCTGCT GCTGCTGCTG CTGCTGCTGA CCCGTCGTCC ATCCAACCCA CCGCTGCACC?2315
CAATTTTTTG AACCCATATA TAGCGAAGCA TCTTCTTGTA CCTAAAAAAA AAAAAAAA ?2373
<210>2
<211>696
<212>PRT
< 213>sugarcane (Saccharum officinarum)
<400>2
Met?Glu?Thr?Arg?Asp?Thr?Thr?Ala?Pro?Leu?Pro?Tyr?Ser?Tyr?Thr
5 10 15
Pro?Leu?Pro?Ala?Ala?Asp?Ala?Ala?Ser?Ala?Glu?Val?Thr?Gly?Thr
20 25 30
Gly?Gly?Ser?Arg?Ser?Arg?Arg?Arg?Arg?Pro?Leu?Cys?Ala?Ala?Ala
35 40 45
Leu?Val?Leu?Ser?Ala?Ala?Leu?Leu?Leu?Ala?Val?Ala?Ala?Leu?Ala
50 55 60
Gly?Val?Gly?Ser?Arg?Val?Ala?Ala?Val?Val?Pro?Arg?Pro?Thr?Thr
65 70 75
Ala?Val?Gly?Glu?Thr?Ala?Gly?Val?Gly?Val?Gly?Val?Val?Pro?Gly
80 85 ?90
Ala?Gly?Thr?Pro?Gln?Ala?Thr?Ser?Thr?Arg?Ser?Arg?Ser?Arg?Gly
95 100 105
Pro?Asp?Ala?Gly?Val?Ser?Glu?Lys?Thr?Ser?Gly?Val?Trp?Thr?Gly
110 115 120
Val?Ile?Asp?Asp?Gly?Ala?Arg?Leu?Arg?Thr?Asp?Ala?Gly?Gly?Asn
125 130 135
Ala?Phe?Pro?Trp?Ser?Asn?Ala?Met?Leu?Gln?Trp?Gln?Arg?Thr?Gly
140 145 150
Phe?His?Phe?Gln?Pro?Gln?Arg?Asn?Trp?Met?Asn?Asp?Pro?Asn?Gly
155 160 165
Pro?Val?Tyr?Tyr?Lys?Gly?Trp?Tyr?His?Leu?Phe?Tyr?Gln?Tyr?Asn
170 175 180
Pro?Asp?Gly?Ala?Ile?Trp?Gly?Asn?Lys?Ile?Ala?Trp?Gly?His?Ala
185 190 195
Val?Ser?Arg?Asp?Leu?Ile?His?Trp?Arg?His?Leu?Pro?Leu?Ala?Met
200 205 210
Leu?Pro?Asp?Gln?Trp?Tyr?Asp?Thr?Asn?Gly?Val?Trp?Thr?Gly?Ser
215 220 225
Ala?Thr?Thr?Leu?Pro?Asp?Gly?Arg?Leu?Ala?Met?Leu?Tyr?Thr?Gly
230 235 240
Ser?Thr?Asn?Thr?Ser?Val?Gln?Val?Gln?Cys?Leu?Ala?Val?Pro?Ala
245 250 255
Asp?Asp?Asp?Asp?Pro?Leu?Leu?Thr?Asn?Trp?Thr?Lys?Tyr?Glu?Gly
260 265 270
Asn?Pro?Ala?Leu?Tyr?Pro?Pro?Pro?Gly?Ile?Gly?Pro?Arg?Asp?Phe
275 280 285
Arg?Asp?Pro?Thr?Thr?Ala?Trp?Phe?Asp?Pro?Ser?Asp?Ser?Thr?Trp
290 295 300
Arg?Ile?Val?Ile?Gly?Ser?Lys?Asp?Asp?Ala?Glu?Gly?Asp?His?Ala
305 310 315
Gly?Ile?Ala?Val?Val?Tyr?Arg?Thr?Arg?Asp?Phe?Val?His?Phe?Glu
320 325 330
Leu?Leu?Pro?Asp?Leu?Leu?His?Arg?Val?Ala?Gly?Thr?Gly?Met?Trp
335 340 345
Glu?Cys?Ile?Asp?Phe?Tyr?Pro?Val?Ala?Thr?Arg?Gly?Lys?Ala?Ser
350 355 360
Gly?Asn?Gly?Val?Asp?Met?Ser?Asp?Ala?Leu?Ala?Lys?Asn?Gly?Ala
365 370 375
Val?Val?Gly?Asp?Val?Val?His?Val?Met?Lys?Ala?Ser?Met?Asp?Asp
380 385 390
Asp?Arg?His?Asp?Tyr?Tyr?Ala?Leu?Gly?Arg?Tyr?Asp?Ala?Ala?Ala
395 400 405
Asn?Ala?Trp?Thr?Pro?Leu?Asp?Ala?Glu?Lys?Asp?Val?Gly?Thr?Gly
410 415 420
Leu?Arg?Tyr?Asp?Trp?Gly?Lys?Phe?Tyr?Ala?Ser?Lys?Thr?Phe?Tyr
425 430 435
Asp?Pro?Ala?Lys?Arg?Arg?Arg?Val?Leu?Trp?Gly?Trp?Val?Gly?Glu
440 445 450
Thr?Asp?Ser?Glu?Arg?Ala?Asp?Val?Ser?Lys?Gly?Trp?Ala?Ser?Leu
455 460 465
Gln?Gly?Ile?Pro?Arg?Thr?Val?Leu?Leu?Asp?Thr?Lys?Thr?Gly?Ser
470 475 480
Asn?Leu?Leu?Gln?Trp?Pro?Val?Glu?Glu?Val?Glu?Thr?Leu?Arg?Thr
485 490 495
Asn?Ser?Thr?Asp?Leu?Ser?Gly?Ile?Thr?Ile?Asp?Tyr?Gly?Ser?Thr
500 505 510
Phe?Pro?Leu?Asn?Leu?Arg?Arg?Ala?Thr?Gln?Leu?Asp?Ile?Glu?Ala
515 520 525
Glu?Phe?Glu?Leu?Asp?Arg?Arg?Ala?Val?Met?Ser?Leu?Asn?Glu?Ala
530 535 540
Asp?Val?Gly?Tyr?Asn?Cys?Ser?Thr?Ser?Gly?Gly?Ala?Ala?Ala?Arg
545 550 555
Gly?Ala?Leu?Gly?Pro?Phe?Gly?Leu?Leu?Val?Leu?Thr?Asp?Lys?His
560 565 570
Leu?His?Glu?Gln?Thr?Ala?Val?Tyr?Phe?Tyr?Val?Ala?Lys?Gly?Leu
575 580 585
Asp?Gly?Ser?Leu?Thr?Thr?His?Phe?Cys?Gln?Asp?Glu?Ser?Arg?Ser
590 595 600
Ser?Ser?Ala?Asn?Asp?Ile?Val?Lys?Arg?Val?Val?Gly?Ser?Ala?Val
605 610 615
Pro?Val?Leu?Glu?Asp?Glu?Thr?Thr?Leu?Ser?Leu?Arg?Val?Leu?Val
620 625 630
Asp?His?Ser?Ile?Val?Glu?Ser?Phe?Ala?Gln?Gly?Gly?Arg?Ser?Thr
635 640 645
Ala?Thr?Ser?Arg?Val?Tyr?Pro?Thr?Lys?Ala?Ile?Tyr?Ala?Asn?Ala
650 655 660
Gly?Val?Phe?Leu?Phe?Asn?Asn?Ala?Thr?Ala?Ala?Arg?Val?Thr?Ala
665 670 675
Lys?Lys?Leu?Val?Val?His?Glu?Met?Asp?Ser?Ser?Tyr?Asn?His?Asp
680 685 690
Tyr?Met?Val?Thr?Asp?Ile
695?696
Claims (7)
1. sugarcane GH32 family soluble acid invertase (SoINV) gene and protein sequence thereof; It is characterized in that: all plant soluble acid invertase full length gene sequences are carried out on the basis of evolutionary analysis; Compare the soluble acid invertase gene cDNA sequence of same family, and the segmental primer of design amplification SoINV gene core, total RNA extracted from the sugarcane young leaflet tablet; And through reverse transcription, with conventional PCR method amplification SoINV gene core fragment; Some in core fragment sequence 5 ' end design then to special primer; At core fragment sequence a 3 ' end special primer of design and two anchor primers; Through RACE round pcr 5 of SoINV gene core district ' three different sequence fragments of end and the 3 ' sequence fragment of end that increases respectively; Three 5 ' terminal sequences are spliced with core area, 3 ' terminal sequence and AY302083 fragment respectively; Obtain sugarcane GH32 three soluble acid invertases of family (SoINV) full length gene cDNA sequence, be designated as SoINV1, SoINV2 and SoINV3 respectively.
2. sugarcane GH32 family's soluble acid invertase (SoINV) gene according to claim 1 and protein sequence thereof is characterized in that: described SoINV1 length overall 2387bp; Through Vector NTI Advance 11 software analysis, the ORF of this sequence is 2055bp, 685 amino acid of encoding; Initiator codon (ATG) is positioned at 215bp place behind the transcription initiation site, and terminator codon (TAG) is positioned at 2272bp, also has the non-coding sequence of one section 115bp thereafter, and has the typical polyA tail of eukaryote; The proteic aminoacid sequence of the sugarcane GH32 SoINV1 of family coded by said gene:
METRDTTAPLPYSYTPLPAADAASAEVTGTGGRSRRRSLCAAALVLSAALLLAVAALAAAGRRPTTAVGETAGVGVVPGVGTPQATSTRSISRGPDAGVSEKTSGAWSGVVDDGGRLRADGGGNAFPWSNAMLQWQRTGFHFQPQRNWMNDPNGPVYYKGWYHLFYQYNPDGAIWGNKIAWGHAVSRDLIHWRHLPLAMLPDQWYDTNGVWTGSATTLPDGRLAMLYTGSTNTSVQVQCLAVPADDDDPLLTNWTKYEGNPALYPPPGIGPRDFRDPTTAWFDPSDSTWRIVIGSKDDAEGDHAGIAVVYRTRDFVHFELLPDLLHRVAGTGMWECIDFYPVATRGKASGNGVDMSDALAKNGAVVGDVVHVMKASMDDDRHDYYALGRYDAAANAWTPLDAEKDVGTGLRYDWGKFYASKTFYDPAKRRRVLWGWVGETDSERADVSKGWASLQGIPRTVLLDTKTGSNLLQWPVEEVETLRTNSTDLSGITIDYGSTFPLNLRRATQLDIEAEFELDRRAVMSLNEADVGYNCSTSGGAAARGALGPFGLLVLTDKHLHEQTAVYFYVAKGLDGSLTTHFCQDESRSSSANDIVKRVVGSAVPVLEDETTLSLRVLVDHSIVESFAQGGRSTATSRVYPTKAIYANAGVFLFNNATAARVTAKKLVVHEMDSSYNHDYMVTDI。
3. sugarcane GH32 family's soluble acid invertase (SoINV) gene according to claim 1 and protein sequence thereof is characterized in that: described SoINV2 length overall 2429bp; Through Vector NTI Advance 11 software analysis, the ORF of this sequence is 2085bp, 695 amino acid of encoding; Initiator codon (ATG) is positioned at 227bp place behind the transcription initiation site, and terminator codon (TAA) is positioned at 2314bp, also has the non-coding sequence of one section 115bp thereafter, and has the typical polyA tail of eukaryote; The proteic aminoacid sequence of the sugarcane GH32 SoINV2 of family coded by said gene:
METRDTTAPLPYSYTPLPAADAASAEVTGTGHRGGGRSRRSSLCAAALVLSAALLLAVAALAGVGGRVAVVPRPTTAVGETAGVGVGPGAGTPQATSTRSISRGPDAGVSEKTSGAWSGVVDDGGRLRADGGGNAFPWSNAMLQWQRTGFHFQPQRNWMNDPNGPVYYKGWYHLFYQYNPDGAIWGNKIAWGHAVSRDLIHWRHLPLAMLPDQWYDTNGVWTGSATTLPDGRLAMLYTGS?TNTSVQVQCLAVPAGDDDPLLTNWTKYEGNPALYPPPGIGPRDFRDPTTAWFDPSDSTWRIVIGSKDDAEGDHAGIAVVYRTRDFVHFELLPDLLHRVAGTGMWECIDFYPVATRGKASGNGVDMSDALAKNGAVVGDVVHVMKASMDDDRHDYYALGRYDAAANAWTPLDAEKDVGTGLRYDWGKFYASKTFYDPAKRRRVLWGWVGETDSERADVSKGWASLQGIPRTVLLDTKTGSNLLQWPVEEVETLRTNSTDLSGITIDYGSTFPLNLRRATQLDIEAEFELDRRAVMSLNEADVGYNCSTSGGAAARGALGPFGLLVLTDKHLHEQTAVYFYVAKGLDGSLTTHFCQDESRSSSANDIVKRVVGSAVPVLEDETTLSLRVLVDHSIVESFAQGGRSTATSRVYPTKAIYANAGVFLFNNATAARVTAKKLVVHEMDSSYNHDYMVTDI。
4. sugarcane GH32 family's soluble acid invertase (SoINV) gene according to claim 1 and protein sequence thereof is characterized in that: described SoINV3 length overall 2373bp; Through Vector NTI Advance 11 software analysis, the ORF of this sequence is 2088bp, 696 amino acid of encoding; Initiator codon (ATG) is positioned at 168bp place behind the transcription initiation site, and terminator codon (TAA) is positioned at 2258bp, also has the non-coding sequence of one section 115bp thereafter, and has the typical polyA tail of eukaryote; The proteic aminoacid sequence of the sugarcane GH32 SoINV3 of family coded by said gene is:
METRDTTAPLPYSYTPLPAADAASAEVTGTGGSRSRRRRPLCAAALVLSAALLLAVAALAGVGSRVAAVVPRPTTAVGETAGVGVGVVPGAGTPQATSTRSRSRGPDAGVSEKTSGVWTGVIDDGARLRTDAGGNAFPWSNAMLQWQRTGFHFQPQRNWMNDPNGPVYYKGWYHLFYQYNPDGAIWGNKIAWGHAVSRDLIHWRHLPLAMLPDQWYDTNGVWTGSATTLPDGRLAMLYTGSTNTSVQVQCLAVPADDDDPLLTNWTKYEGNPALYPPPGIGPRDFRDPTTAWFDPSDSTWRIVIGSKDDAEGDHAGIAVVYRTRDFVHFELLPDLLHRVAGTGMWECIDFYPVATRGKASGNGVDMSDALAKNGAVVGDVVHVMKASMDDDRHDYYALGRYDAAANAWTPLDAEKDVGTGLRYDWGKFYASKTFYDPAKRRRVLWGWVGETDSERADVSKGWASLQGIPRTVLLDTKTGSNLLQWPVEEVETLRTNSTDLSGITIDYGSTFPLNLRRATQLDIEAEFELDRRAVMSLNEADVGYNCSTSGGAAARGALGPFGLLVLTDKHLHEQTAVYFYVAKGLDGSLTTHFCQDESRSSSANDIVKRVVGSAVPVLEDETTLSLRVLVDHSIVESFAQGGRSTATSRVYPTKAIYANAGVFLFNNATAARVTAKKLVVHEMDSSYNHDYMVTDI。
5. sugarcane GH32 family's soluble acid invertase (SoINV) gene according to claim 1 and protein sequence thereof; It is characterized in that: described sugarcane GH32 three soluble acid invertases of family (SoINV) full length gene cDNA sequence, core area sequence (M-SoINV) is to use following nucleotide sequence to obtain as primer amplification:
N?f1：5′-tctggggcaacaagatcgcgt-3′；
N?r1：5′-aaattgggtgcagcggtgggt-3′。
6. sugarcane GH32 family's soluble acid invertase (SoINV) gene according to claim 1 and protein sequence thereof; It is characterized in that: the sequence of described 3 ' end (3 '-SoINV) be to use following nucleotide sequence to be primer; With 3 '-primer that Full RACEKit (TAKARA) provides combines, and the amplification of Bird's Nest formula obtains:
GZ?f2：5′-cctcttcaacaacgccaccgccg-3′。
7. sugarcane GH32 family's soluble acid invertase (SoINV) gene according to claim 1 and protein sequence thereof; It is characterized in that: described 5 ' terminal sequence (5 '-SoINV) be respectively to use following nucleotide sequence to be primer; With 5 '-primer that Full RACE Kit (TAKARA) provides combines, and the amplification of Bird's Nest formula obtains:
The primer of the soluble acid invertase SoINV1 of sugarcane GH32 family gene 5 ' end is used to increase:
INVSYR1：(5′-gttggtggagccggtgtagagcat-3′)；
INVR1：(5′-gcccctgctgatgctcctggtcg-3′)；
The primer of the soluble acid invertase SoINV2 of sugarcane GH32 family gene 5 ' end is used to increase:
INVSY?R1：(5′-gttggtggagccggtgtagagcat-3′)；
INVR2：(5′-acgtcgcctgtggtgtcccc-3′)；
The primer of the soluble acid invertase SoINV3 of sugarcane GH32 family gene 5 ' end is used to increase:
INVSY?R1：(5′-gttggtggagccggtgtagagcat-3′)；
INVR3：(5′-ggggtcgttcatccagttcctct-3′)。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011103657488A CN102505021A (en) | 2011-11-17 | 2011-11-17 | Soluble acid invertase (So INV) gene of sugarcane GH32 family and protein sequence thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011103657488A CN102505021A (en) | 2011-11-17 | 2011-11-17 | Soluble acid invertase (So INV) gene of sugarcane GH32 family and protein sequence thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN102505021A true CN102505021A (en) | 2012-06-20 |
Family
ID=46217151
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2011103657488A Pending CN102505021A (en) | 2011-11-17 | 2011-11-17 | Soluble acid invertase (So INV) gene of sugarcane GH32 family and protein sequence thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102505021A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102888414A (en) * | 2012-09-13 | 2013-01-23 | 广西大学 | Sugarcane neutral/alkaline invertase gene and encoded protein sequence thereof |
| CN104805102A (en) * | 2015-05-06 | 2015-07-29 | 中国农业科学院作物科学研究所 | Method for cloning soluble acid invertase gene SAI-1 of sorghum |
| CN105420250A (en) * | 2015-10-10 | 2016-03-23 | 华南农业大学 | Lychee cell wall acid invertase gene and application thereof |
-
2011
- 2011-11-17 CN CN2011103657488A patent/CN102505021A/en active Pending
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102888414A (en) * | 2012-09-13 | 2013-01-23 | 广西大学 | Sugarcane neutral/alkaline invertase gene and encoded protein sequence thereof |
| CN104805102A (en) * | 2015-05-06 | 2015-07-29 | 中国农业科学院作物科学研究所 | Method for cloning soluble acid invertase gene SAI-1 of sorghum |
| CN105420250A (en) * | 2015-10-10 | 2016-03-23 | 华南农业大学 | Lychee cell wall acid invertase gene and application thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| BRPI1002997A2 (en) | transgenic saccharin sorghum with altered lignin composition and process of preparation thereof | |
| CN109694872A (en) | The method of controlling gene expression | |
| CN103333901B (en) | Liriodendron hybrid LhWOX1 gene and application thereof | |
| CN115927029A (en) | Recombinant saccharomyces cerevisiae for producing cannabigerol acid and construction method and application thereof | |
| CN102505021A (en) | Soluble acid invertase (So INV) gene of sugarcane GH32 family and protein sequence thereof | |
| CN101886086B (en) | Trehalose-6-phosphate synthase gene sequence derived from Selaginella pulvinata Maxim and method for cloning same | |
| CN102226184B (en) | Method for cultivating transgenic nitrogen-fixing plants | |
| CN108048469B (en) | The Ravenna grass class Calmodulin Gene ErCML30 expressed in Ravenna grass wild species by low temperature stress | |
| CN107365371B (en) | Sugarcane flowering regulatory protein ScFT-2 and coding gene thereof | |
| CN110846293B (en) | Lysophosphatidic acid acyltransferase | |
| CN110452917B (en) | Wild grape VyGOLS gene and application of encoding protein thereof in drought stress | |
| CN1606623A (en) | Production of stilbenes in transgenic plants and the method of producing thereof | |
| CN113584051B (en) | Application of GhGAI gene in regulation and control of plant flowering | |
| CN108795944A (en) | Cotton long-chain non-coding RNA-lnc973 and its application in plant salt endurance | |
| CN107417782A (en) | Sugarcane florescence modulin ScFT 3 and its encoding gene | |
| CN102021188A (en) | Artemisia sphaerocephala delta 12 fatty acid desaturase (As FAD2) gene and application | |
| CN113337522A (en) | Application of cotton GhNFYC4 gene in promoting plant flowering | |
| CN102586264B (en) | Method for improving plant yield | |
| CN102533817A (en) | Sugarcane cell wall-bound invertase genes and coding protein sequence thereof | |
| CN105969797A (en) | Autofluorescence relevant micromolecule RNA of chloroplast and application of autofluorescence relevant micromolecule RNA | |
| CN106834343B (en) | Sucrose synthase is in the regulation developmental application of fruit | |
| CN104293808A (en) | Liriodendron hybrids LhMKK2 gene and expression protein and application thereof | |
| CN105802976A (en) | Gene for regulating rice resistance to drought and salt/alkali and application thereof | |
| CN110272906A (en) | A kind of Chinese tamarisk salt stress responsive genes TcSBP1 and its miRNA resistance target rTcSBP1 and application | |
| CN105647928B (en) | Potato stu-miR5992a and stu-miR5992b and its application |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20120620 |