CN109182344A - The clone of Aechmea fasciata AfTOE1L gene and its application - Google Patents
The clone of Aechmea fasciata AfTOE1L gene and its application Download PDFInfo
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- CN109182344A CN109182344A CN201810964319.4A CN201810964319A CN109182344A CN 109182344 A CN109182344 A CN 109182344A CN 201810964319 A CN201810964319 A CN 201810964319A CN 109182344 A CN109182344 A CN 109182344A
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Abstract
The present invention relates to a kind of clone of Aechmea fasciata AfTOE1L gene and its applications, specifically Aechmea fasciata AP2/EREBP transcription factor AfTOE1L encoding gene and its plant expression vector.Aechmea fasciata lobus cardiacus total serum IgE is extracted, reverse transcription synthesizes cDNA, as template, designs special primer, the full-length cDNA of AfTOE1L gene is obtained using RACE technology, and obtain its CDS sequence on this basis.The CDS sequence is connected on plant expression vector Cam35S-gfp, a new plant expression vector is constructed.To arabidopsis transgenic line research shows that the gene is related with the regulation of flowering time and seed size, therefore, the acquisition of AfTOE1L gene, it blooms for artificial finely regulating bromelia and cultivates pineapple new varieties by genetic engineering means and lay a good foundation, there is important theory and practice significance.
Description
Technical field
The invention belongs to molecular biology and field of biotechnology, it is related to APETALA2/ethylene- in Aechmea fasciata
Responsive element binding protein (AP2/EREBP) transcription factor family member's encoding gene AfTOE1-
Clone, recombination and the split floral shape of like (AfTOE1L) determines and the analysis and application of seed size adjusting function.
Background technique
AP2/EREBP transcription factor is a kind of family's egg being widely present in plant, protist, cyanobacteria and bacteriophage
It is white, it plays an important role in organism reply external source environmental signal regulation growth and development process.AP2/EREBP transcription factor man
Race's albumen can be divided into AP2 subtribe and EREBP subtribe two major classes, and wherein AP2 subtribe member contains 2 AP2 structural domains, EREBP
Subtribe then contains only an AP2 structural domain.AP2 structural domain is highly conserved, is made of about 68 amino acid residues, wherein by 18
The nucleus of amino acid composition constitutes hydrophilic α spiral.AP2 structural domain plays the function of AP2 subtribe albumen heavy to closing
The effect wanted.In addition to containing there are two other than conservative AP2 structural domain, most of AP2 subtribes also contain a nuclear localization signal.
Have 6 AP2 subtribe members in arabidopsis, respectively AP2, TARGET OF EAT 1 (TOE1), TOE2, TOE3,(SMZ) and SCHNARCHZAPFEN (SNZ).Wherein AP2 is in seed development, stem cell function
It maintains, bloom starting and floral organ decision etc. all have critical function.For example, AP2 both can be used as transcriptional activation because
Son regulates and controls the expression of repressor AGAMOUS-LIKE15 (AGL15) of blooming, and can be used as a transcription inhibitory factor inhibition and open
Flower activates the expression of sub- SUPPRESSOR OF OVEREXPRESSION OF CONSTANS1 (SOC1), so that regulation has been bloomed
Begin.In addition, as A genoid, AP2 and APETALA1 (AP1) and E genoid are total in the ABCE model that floral organ determines
With the development for having regulated and controled first round calyx and the second wheel petal.In addition, in flowering period conversion, AP2 in arabidopsis and
Other 5 AP2 subtribe member's functional redundancies, in addition to TOE3, being overexpressed other 5 AP2 genoids can significant late blooming.It is quasi-
The southern significant early blossoming of mustard toe1 mutant, toe1toe2 double-mutant flowering time further shift to an earlier date.Although smz or snz mutation
Body flowering time does not change, but four flowering times of the prominent toe1toe2smzsnz than double-mutant toe1toe2 are earlier.This
Outside, the Arabidopsis plant after the mutation of all AP2 subtribe encoding genes relative to four prominent toe1toe2smzsnz flowering time then
Again in advance.The study found that the promoter region that TOE1 and SMZ can be bonded directly to florigen FT inhibits the latter to express, thus
Inhibition is bloomed；AP2 then can be incorporated into AG gene second includes on the AT-rich element of sub-district, determines colored form.
Aechmea fasciata is a kind of very strong sight flower foliage plant of ornamental value, and Aechmea fasciata is mainly at present with flowering naturally
It is main, how the artificial finely regulating florescence, so that pineapple is bloomed in market fast-selling period, and new by genetic engineering means cultivation pineapple
Kind has become one of research direction of researcher.The AP2 subtribe member of AfTOE1L and other plant in Aechmea fasciata are high
Spend homologous, two typical AP2 structural domains and nuclear localization signal are also all very conservative, thus it is speculated that may rise in florescence control important
Effect.
Summary of the invention
The object of the present invention is to provide a kind of clone of Aechmea fasciata AfTOE1L gene and its applications, with the Aechmea fasciata heart
Leaf is material, extracts total serum IgE using CTAB method and is respectively synthesized 5 ' and 3 ' RACE templates, obtains Aechmea fasciata through PCR amplification
AfTOE1L gene.The CDS sequence construct of Aechmea fasciata AfTOE1L gene is transferred to arabidopsis, heterologous table under 35S promoter
Seed up to the arabidopsis transgenic line of AfTOE1L becomes smaller, mass of 1000 kernel decline, and significant evening flower under the conditions of short-day.
Aechmea fasciata AfTOE1L gene provided by the present invention, gene order have SEQ ID NO:1 institute in sequence table
The nucleotide sequence shown, cDNA sequence overall length 2160bp, by the 5 ' noncoding regions of 606bp, 150bp 3 ' noncoding regions and
The open reading frame of 1404bp forms.The albumen of the gene one 467 amino acid residue of coding, molecular weight 51.32kDa, etc.
Electricity point is 6.65.
The clone of above-mentioned Aechmea fasciata AfTOE1L gene:
Using Aechmea fasciata lobus cardiacus as material, total serum IgE is extracted using CTAB method and is respectively synthesized 5 ' and 3 ' RACE templates.It is obtaining
In the transcript profile sequence basis obtained, the special interior Outside primer at amplification 5 ' end and 3 ' terminal sequences is separately designed, primer sequence is as follows:
5 ' end Outside primers: 5 '-GGTTACAGCCTCCCTCCCATTACATT-3 '
5 ' end inner primers: 5 '-TGCCCAGAAACTGTCCCATCCG-3 '
3 ' end Outside primers: 5 '-CAGGTGAAGAAGAGTAGGAGGGGGC-3 '
3 ' end inner primers: 5 '-GAAGCTCGGATGGGACAGTTTCTG-3 '
5 ' the ends come and 3 ' terminal sequences are amplified after sequence verification with transcript profile sequence, finally obtain AfTOE1L gene
The cDNA sequence of overall length 2160bp.
AP2 member in AfTOE1L protein sequence and a variety of species is subjected to phylogenetic analysis, find its in arabidopsis
AtTOE1 parent source relationship in 6 AP2 subtribe members is nearest, implies that the two may have similar function (Fig. 1).
According to the CDS sequence of the AfTOE1L gene of acquisition, inventor devises special primer and has expanded its DNA sequence dna.Expand
The primer for increasing DNA sequence dna is as follows:
5 ' end primers: 5 '-ATGATGCTCGATCTGAACATGGCCG-3 '
3 ' end primers: 5 '-TCAGCTCTTGAAGAAATTCAAAGAA-3 '
The DNA sequence dna overall length 3498bp of the gene, is made of, the AtTOE1 with arabidopsis 8 exons and 7 intrones
Exon it is identical with introne number (Fig. 2).
6 AP2 family members of AfTOE1L protein sequence and arabidopsis are subjected to sequence analysis, discovery AfTOE1L tool
There are 2 conservative AP2 structural domains and 1 highly conserved nuclear localization signal sequence, shows that AfTOE1L albumen is Aechmea fasciata
In AP2 subtribe member (Fig. 3).
According to the CDS sequence of the AfTOE1L of acquisition, design adds KpnI and XbaI enzyme cutting site at 5 ' ends and 3 ' ends respectively
Primer, sequence is as follows:
It is overexpressed primers F:
5’-GGGGTACCATGATGCTCGATCTGAACAT-3’
It is overexpressed primer R:
5’-GCTCTAGATCAGCTCTTGAAGAAATTC-3’
By PCR amplification, the CDS sequence with restriction enzyme site is obtained.Sequence is by KpnI and XbaI double digestion and same pair
Expression vector Cam35S-gfp connection after digestion, finally obtains over-express vector.
After the expression vector Cam35S-gfp with AfTOE1L CDS sequence built is transferred to Agrobacterium EHA105,
Disseminate arabidopsis floral.The T0 of acquisition is sowed for seed and is carried out on the solid plate of MS+2% sucrose+50mg/mL hygromycin
Resistance screening obtains T1 for transgenic positive plant.The seed in T1 generation continues to be seeded on resistance plate, obtains T2 for transgenosis sun
Property plant.In T2 generation, continues to sow, by observing T3 represents whether type separates and molecular level identifies whether be homozygote plant.It sees
The seed for examining the transgenic positive strain in discovery T3 generation becomes smaller (Fig. 4 A, B, C, D, E), and mass of 1000 kernel declines (Fig. 4 F), and in short day
The significant evening flower (Fig. 4 G) according under the conditions of.
For the present invention using Aechmea fasciata lobus cardiacus as material, clone obtains Aechmea fasciata AfTOE1L gene, phylogenetic analysis result
Show that AfTOE1L and the AtTOE1 parent source relationship in arabidopsis in 6 AP2 members are nearest.Gene order structural analysis it has also been found that
AfTOE1L is similar with AtTOE1, all has 8 exons and 7 intrones.Further experimental result confirms that AfTOE1L is
It is positioned at the transcription activator of nucleus.The CDS sequence construct of AfTOE1L is transferred to arabidopsis, heterologous table under 35S promoter
Arabidopsis transgenic line seed up to AfTOE1L becomes smaller, mass of 1000 kernel decline, and the evening flower under short-day, shows that AfTOE1L is
Regulate and control an important transcription factor of Aechmea fasciata seed development and flowering time, blooms for artificial finely regulating bromelia
And cultivate pineapple new varieties by genetic engineering means and lay a good foundation, there is important theory and practice significance.
Detailed description of the invention
Fig. 1 is the phylogenetic analysis of Aechmea fasciata AfTOE1L Yu other AP2 subtribe albumen.
Fig. 2 is Aechmea fasciata AfTOE1L and arabidopsis AP2 subtribe encoding gene structure chart
Fig. 3 is the sequence analysis of Aechmea fasciata AfTOE1L, AfAP2-1 and 6 AP2 subtribe members of arabidopsis.
It is respectively arabidopsis Col-0 that Fig. 4, which is 35S::AfTOE1L arabidopsis transgenic line phenotype (A), (B), (C) (D),
Wild type, the Col-0 type arabidopsis for turning empty carrier Cam35S-gfp, No. 77 strains 35S::AfTOE1L arabidopsis transgenic line
The 35S::AfTOE1L arabidopsis transgenic line drawing of seeds of system and No. 157 strains；It (E) is seed size statistical result；(F) it is
The statistical result of thousand grain weigth；It (G) is (8h light/16h is dark) 35S::AfTOE1L arabidopsis transgenic line under the conditions of short-day
The evening flower phenotypic map of system.
Specific embodiment
With reference to embodiment, the embodiment of the present invention is furthur described in detail.Following embodiment is used for
Illustrate the present invention, but is not intended to limit the scope of the invention.In the following examples, the experimental methods for specific conditions are not specified, usually
According to normal conditions, or according to the normal condition proposed by manufacturer.
Embodiment one: the clone of Aechmea fasciata AfTOE1L gene
1, the extraction of total serum IgE
(1) 850 μ L CTAB extracting solutions are taken to be dispensed into 2mL centrifuge tube, 65 DEG C of preheatings；
(2) 0.2g material is weighed, the last preheating for being transferred to added 2% mercaptoethanol immediately of pulverizing in liquid nitrogen
In the centrifuge tube of CTAB extracting solution, vortex mixed, 65 DEG C of warm bath 6min；
(3) isometric chloroform/isoamyl alcohol (24:1), vortex mixed, room temperature, 10000rpm centrifugation 15min is added；
(4) supernatant is carefully drawn into a new 2mL centrifuge tube, repeats step 3；
(5) supernatant is carefully drawn into a new 1.5mL centrifuge tube, and the 8mol/L LiCl of 1/3 volume, 4 DEG C of precipitatings are added
Overnight；
(6) 4 DEG C, 10000rpm is centrifuged 30min；
(7) supernatant is abandoned, precipitating is washed with 70% ethyl alcohol and dehydrated alcohol respectively, with 20 μ L RNase-free water after drying
Dissolution, solution is the RNA of high-purity.
2, the clone of AfTOE1L full-length cDNA
(1) it is first depending on the SMARTer of ClontechTMRACE cDNA Amplifiation Kit is respectively synthesized 5 ' ends
Template and 3 ' Side Templates, then it is utilized respectively 5 ' end Outside primers and inner primer 5 ' terminal sequences of amplification；Using 3 ' end outside and it is interior
3 ' terminal sequence of side primer amplification.The pcr amplification product of acquisition is connected to pEASY-T1 cloning vector, converts Escherichia coli, warp
Sequencing is carried out after PCR identification.It measures correct sequence to be spliced with original transcript profile sequence, obtains AfTOE1L overall length
CDNA sequence.
(2) according to the above splicing sequence design full-length cDNA amplification primer:
5 ' end primers: 5 '-GGGACACTCTCCTCCTCTCTCTATA-3 '
3 ' end primers: 5 '-ACCAAGAATTATTCCATTCACCC-3 '
According to the T of primermAfter value carries out the optimization of PCR condition, expanded according to following procedure:
PCR system and condition are as follows:
The PCR product of acquisition is purified, purifying uses the Ago-Gel QIAquick Gel Extraction Kit of OMEGA, according to specification into
Row.Product after purification is connect with pEASY-blunt carrier, converts Escherichia coli.With the cDNA amplimer of AfTOE1L to list
Bacterium colony carries out PCR detection, has the clone of purpose band to send sequencing.
3, the extraction of total DNA
(1) it weighs the tender Aechmea fasciata lobus cardiacus blade of 0.2g to enter in 1.5-mL centrifuge tube, be freezed in liquid nitrogen, dismembyator is ground
It clays into power.
(2) it is rapidly added the CTAB lysate (with 2% mercaptoethanol of preceding addition) of 65 DEG C of 500 μ L preheatings, mixing is placed on
65 DEG C of warm bath 30min, mix by phased manner therebetween, so that cracking is abundant.
(3) after room temperature is cooling, isometric phenol/chloroform (1:1) is added, 5min, the room 12000rpm are stood after being mixed by inversion
Temperature centrifugation 15min.
(4) it carefully draws supernatant to enter in new centrifuge tube, isometric chloroform is added, be mixed by inversion to extract.It stands
5min.12000rpm room temperature is centrifuged 15min.
(5) it carefully draws supernatant to enter in new centrifuge tube, the NaAc solution of 1/3 volume 3mol/L is added, mixes.
(6) dehydrated alcohol of the pre-cooling of 2 times of volumes is added, is mixed by inversion and is placed on -20 DEG C of precipitating at least 20min.
(7) 4 DEG C, 12000rpm is centrifuged 15min, abandons supernatant.
(8) suitable 70% ethanol washing 2 or 3 times are added, pay attention to not outwelling white precipitate.
(9) centrifuge tube after washing is inverted on blotting paper and is dried.
(10) DNA of the ddH2O dissolution tube bottom of 20 μ L is added.Next experiment can directly be carried out or be placed in -20 DEG C
It freezes.
4, the clone of AfTOE1L gene order
According to the cDNA sequence of the AfTOE1L gene obtained, the ORFfinder online software of NCBI is utilized
(http://www.ncbi.nlm.nih.gov/orffinder/) predicts the CDS sequence of AfTOE1L.It is set according to CDS sequence
Count primer, using the DNA of extraction as template, PCR amplification full length gene.Primer sequence is as follows:
5 ' end primers: 5 '-ATGATGCTCGATCTGAACATGGCCG-3 '
3 ' end primers: 5 '-TCAGCTCTTGAAGAAATTCAAAGAA-3 '
According to the T of primermAfter value carries out the optimization of PCR condition, expanded according to following procedure:
PCR system and condition are as follows:
The PCR product of acquisition is purified, purifying uses the Ago-Gel QIAquick Gel Extraction Kit of OMEGA, according to specification into
Row.Product after purification is connect with pEASY-blunt carrier, converts Escherichia coli.With the CDS amplimer of AfTOE1L to list
Bacterium colony carries out PCR detection, has the clone of purpose band to send sequencing.
Embodiment two: the building of Aechmea fasciata AfTOE1L gene plant expression vector
1, according to the CDS sequence of Aechmea fasciata AfTOE1L, referring to plant expression vector Cam35S-gfp map, selection is closed
Suitable restriction enzyme site design primer.Primer sequence is as follows:
It is overexpressed primers F:
5’-GGGGTACCATGATGCTCGATCTGAACAT-3’
It is overexpressed primer R:
5’-GCTCTAGATCAGCTCTTGAAGAAATTC-3’
Using the cDNA of total serum IgE reverse transcription as template, PCR amplification is carried out.Body of the PCR reaction system with amplification full-length gene
System, in addition to annealing temperature becomes 62 DEG C from 60 DEG C, other reaction conditions are all the same.
2, the PCR product of acquisition is purified, purifying uses the Ago-Gel QIAquick Gel Extraction Kit of OMEGA, according to specification
It carries out.Product after purification is connect with pEASY-blunt carrier, converts Escherichia coli.Method for transformation is carried according to pEASY-blunt
Body specification carries out.PCR detection is carried out to single colonie with the expression vector amplimer of AfTOE1L, there is the clone of purpose band
Send sequencing.
3, plasmid extraction is carried out to the correct monoclonal of sequencing result.It should with two restriction enzymes of KpnI and XbaI
Gene is cut from pEASY-blunt carrier, is connect with the Cam35S-gfp of same enzyme digestion.It is thin that connection product converts DH5 α
Then born of the same parents cultivate on the LB solid plate of the kanamycins containing 100mg/L, the digestion of PCR identification and Plasmid DNA is carried out to bacterium colony
Analysis.The recon built is named as 35S::AfTOE1L.
The Function Identification of embodiment three: AfTOE1L
1, (23 DEG C, 16h light/8h is dark, 120 μm of ol m of light intensity between culture-2s-1) plantation arabidopsis Col-0 wild type plant
Strain is simultaneously bloomed to it.
2, expression vector 35S::AfTOE1L is converted into Agrobacterium EHA105, the Agrobacterium monoclonal that picking has been identified is in containing
In the LB liquid medium of 50mg/L kanamycins and 100mg/L rifampin, 28 DEG C of shaken cultivations.
3,5 minutes collection Agrobacteriums of 5000rpm room temperature centrifugation, and precipitating conversion fluid (5% sucrose, 0.02%Silwet, 1
× MS) it is suspended into OD600It is 0.8~1.0.
4, the inflorescence that the previous day is watered with to the plant of water is immersed in 20~50sec in the outstanding conversion fluid for having thallus, is protected from light training
It supports for 24 hours, moves under dim light and grow later.
5, it so converts 3~5 times, collects the mature seed of plant.
6, the seed 10min collected is sterilized using 0.1% mercuric chloride, then point is multicast to the MS solid of the hygromycin containing 50mg/L
Resistance screening is carried out on plating medium, obtains T1 for positive transgenic plant.
7, T1 is collected for seed, is carried out resistance screening on the MS solid plate culture medium of 50mg/L hygromycin again, is obtained
The T2 of trait segregation is obtained for positive transgenic plant.
8, in T2 generation, continues to sow, and represents whether type separates and molecular level identifies whether be that homozygote is planted by observing T3
Strain.The seed of the transgenic positive strain in observation discovery T3 generation becomes smaller, mass of 1000 kernel decline, and significant evening under the conditions of short-day
Flower.
The above is only a preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For member, without departing from the technical principles of the invention, several improvements and modifications can also be made, these improvements and modifications
Also it should be regarded as protection scope of the present invention.
Sequence table
<110>Tropical Crop Variety Resource Institute of Chinese Academy of Tropical Agricultural Sciences
<120>clone of Aechmea fasciata AfTOE1L gene and its application
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 2186
<212> DNA
<213>Aechmea fasciata (Aechmea fasciata)
<400> 1
gggacactct cctcctctct ctatatctct ctctctctct ctctcatctc gttcatcgcc 60
gactctctct tcctttgaga gaactcgtcg ccgactcctt tactgttcgc gacctcgctt 120
ccctcgatta gattgttact atcctcctct tcgtcctcct cttcgtcttc ttcttcttct 180
tctatttgtt tttaacattt tcatcgtcaa tcgactcttc ttggatctga ttgttagggt 240
atttgggatt cgttcatgta gtttttcgat cgatttagct gaggaagggg aggaatcgga 300
ggggggagag gaggaggagg tggtaacttg aggaatttgg aaggattcgt attgattaat 360
gtattggtcg atcgattttt gcttccgagc tcgtaatccg agtccaaatg tttcgtaaga 420
acgactcgtt ttcatcctgc ttctaccggt tcctgctccg gctcttcgtt tttgtagtag 480
agctggggtc gaaaccctag cttgcccgga atccctgatt cagtgtctcg gaacagcgga 540
gccggctccg atcggatctt agtaggctga gtttttgttt tgagttatct tataacgacg 600
aggaggatga tgctcgatct gaacatggcc ggggagccct ccgatttagc ggcgaagccc 660
cggtcgaagg gctgcggcag tgattccgcc acctccgact cctcggtcct caatgccgag 720
gccgggggta atcctgccga tgaggattcg agcactgcac agccgacgac ggctggtatg 780
gccttgcagt tcagtatact gaggagctcg gcgtcggccg agctggagaa cgaggcggag 840
gacgagaact gcggttcccc ggagtccgac gttgtgacca agcagttatt tcccctcaac 900
gcggtctcgg agtggccgca ggcgctgcag acgtcttcgt cgtctaatag gtcgcaattg 960
atcgatctcc cgttctgcca ctccgatgct cagccggaga ttaaggtttt tccgcagccg 1020
cagctgcagc cgcctcagca gcagcaggtg aagaagagta ggagggggcc gcggtcccgg 1080
agctctcagt atagaggggt tacgttctac cgtagaaccg gtagatggga gtcgcacata 1140
tgggactgcg ggaaacaagt ttatttggga ggattcgaca ctgctcatgc tgctgctaga 1200
gcatacgatc gagctgcgat caagtttcga ggtgtcgatg ccgacatcaa tttcaacctt 1260
agtgactatg aggaggatat gaagcagatg aggaatctaa caaaggaaga gtttgtgcac 1320
attctgcgtc ggcaaagcac cgggttctcc agaggtagct caaaatatag aggggttacg 1380
cttcataagt gtggccgctg ggaagctcgg atgggacagt ttctgggcaa gaaggcttat 1440
gacaaggcag ctataaaatg taatgggagg gaggctgtaa ccaattttga tcctaacgtc 1500
tatgatggcg agctacttgc tgatgtcgat gttgaaggtc atgaagttga tttgagcttg 1560
aggatatctc aacctatcat ccaaagtccg aagagggatc atgaatctgt tggcatccag 1620
ttccagtatg gatcgttgga aacatctgaa accaagaaat cgaaggttga cagctccttc 1680
caacaggcta gtcagccaca caacacagtg atgctgcctg agcaccctta ttcccgaact 1740
gcaataggcc ctgccatctt tactgctagt gaggaaagag ccatagagaa gaggccggag 1800
gcgcccggcg cgcaatctct ccccatctgg gcaaggatga cgcatggccg taatactagt 1860
attgctccat tactgccgct gttctcgtct gcagcatcat caggattctc tacacctact 1920
accgtgactg ccatgctcgc caccccgcgc tcttcgaaca cgcatttcca ttcaagccct 1980
gcatcttctt tgaatttctt caagagctga ttctaatgga aaagccaaaa gaggcatgcg 2040
taatgtgtat acacacacct tgatgtttta agccactgta tgtatactat cactacacac 2100
aaacatcaaa ctcgatagct taaaagaagc taatcttggg tgaatggaat aattcttggt 2160
aaaaaaaaaa aaaaaaaaaa aaaaaa 2186
Claims (3)
1. a kind of Aechmea fasciata AfTOE1L gene, it is characterised in that the gene order has in sequence table shown in SEQ ID NO:1
Nucleotide sequence.
2. a kind of expression vector of Aechmea fasciata AfTOE1L gene described in claim 1, it is characterised in that utilize claim
Aechmea fasciata AfTOE1L gene described in 1 is the expression vector of purpose gene.
3. Aechmea fasciata AfTOE1L expression vector according to claim 2, it is characterised in that use claim 1 institute
The Aechmea fasciata AfTOE1L gene stated is inserted on plant expression vector Cam35S-gfp, is built under CaMV35S promoter
Swim the efficient expression vector containing Aechmea fasciata AfTOE1L gene.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201810964319.4A CN109182344A (en) | 2018-08-23 | 2018-08-23 | The clone of Aechmea fasciata AfTOE1L gene and its application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN201810964319.4A CN109182344A (en) | 2018-08-23 | 2018-08-23 | The clone of Aechmea fasciata AfTOE1L gene and its application |
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