CN113151268B - African agapanthus dehydratin ApY 2 SK 2 Gene promoter sequence and use thereof - Google Patents

African agapanthus dehydratin ApY 2 SK 2 Gene promoter sequence and use thereof Download PDF

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CN113151268B
CN113151268B CN202110350763.9A CN202110350763A CN113151268B CN 113151268 B CN113151268 B CN 113151268B CN 202110350763 A CN202110350763 A CN 202110350763A CN 113151268 B CN113151268 B CN 113151268B
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张荻
陈晓童
刘涛
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Shanghai Jiaotong University
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Abstract

The invention discloses a agapanthus dehydratum ApY 2 SK 2 Gene promoter sequence and its use. Which comprises one of the following: (1) a polynucleotide having a nucleotide sequence shown as SEQ ID No. 1; (2) a polynucleotide having a nucleotide sequence which has more than 80% homology, preferably more than 85% homology, more preferably more than 90% homology with the polynucleotide shown in SEQ ID No.1, and having the function of the polynucleotide shown in SEQ ID No. 1. The detection of the activity of GUS enzyme proves that the promoter has the activity of the promoter and is enhanced by PEG, salt, osmosis, low temperature and ABA induction activity, so that the effect of the dehydrin gene on responding to adversity stress in agapanthus is shown.

Description

African agapanthus dehydratin ApY 2 SK 2 Gene promoter sequence and use thereof
Technical Field
The invention relates to the technical field of biology, in particular to agapanthus dehydratum protein ApY 2 SK 2 A promoter for a gene.
Background
Agapanthus praecox (Agapanthus praecox) is a perennial rooted stem flower called "lilium glaucoides" belonging to the genus Agapanthus of the family lycolidaceae. The lotus flower is beautiful, has long florescence and certain stress resistance, is an excellent ornamental plant, is also an important garden flower, can be used as a material for flower arrangement, and is often used as a plant material for pot plants, flower beds and coastal park ground coverings in gardens. Meanwhile, the African agapanthus has certain cultural connotation and enjoys the reputation of 'love flowers' in European and American countries.
Dehydrins are also known as rab (reactive to aba) and belong to the LEA protein family II of late embryonic developmental proteins (LEA proteins). The dehydrated vegetable has strong hydrophilicity and thermal stability. It is widely present in higher plants, algae, yeasts, nematodes and bacteria, and has the functions of stabilizing cell membranes, scavenging free radicals, binding metal ions, protecting cells from dehydration damage, protecting molecular chaperones and DNA/RNA binding. The plant stress resistance has higher correlation with the expression level of the dehydrin in the stress environment, and the expression level of the dehydrin in a resistant plant is higher than that of a sensitive plant. Studies on the regulation of the expression of the dehydrin gene are crucial for the stress resistance of plants.
The promoter is a DNA sequence which is positioned in the upstream region of the upstream 5' -end of the structural gene and used for regulating gene transcription, is a core element of transcription regulation and can respond to abiotic stress and hormone. The analysis of the sequence characteristics of the dehydrin promoter is the key to understanding the transcriptional regulation expression pattern of the gene and its regulation mechanism. At present, researches on the dehydrin promoter mainly focus on plant materials such as arabidopsis thaliana, wheat and bermuda grass, but related documents on the cloning and regulation mode analysis of the dehydrin gene of the agapanthus africanus, which is an ornamental garden flower, are rarely published.
Disclosure of Invention
In view of the above-mentioned disadvantages of the prior art, the present invention aims to fill up the problem of agapanthus dehydratum gene ApY 2 SK 2 The invention also provides a nucleic acid sequence of the dehydrin gene promoter; the invention discloses a agapanthus dehydratum gene ApY 2 SK 2 The promoter activity change mode of the promoter sequence after the arabidopsis is subjected to stress treatment is ApY by utilizing the gene engineering technology in the future 2 SK 2 The method lays a foundation for regulating and controlling the time-space characteristics of gene expression, lays a theoretical foundation for improving the anti-stress capability of the agapanthus and molecular breeding work, and has great application value.
To achieve the above and other related objects, the present invention is achieved by the following aspects.
In a first aspect, the invention provides agapanthus dehydratum ApY 2 SK 2 A gene promoter sequence comprising one of:
(1) the nucleotide sequence is shown as the polynucleotide sequence shown in SEQ ID No. 1;
(2) the polynucleotide sequence has more than 80 percent of homology (preferably more than 85 percent of homology, more preferably more than 90 percent of homology) with the polynucleotide sequence shown in SEQ ID No.1 and has the functions of the polynucleotide shown in SEQ ID No. 1.
The polynucleotide sequence in (2) is specifically a polynucleotide sequence which is obtained by substituting, deleting or adding one or more bases to the polynucleotide sequence shown in SEQ ID No.1 and has the functions of the polynucleotide sequence shown in SEQ ID No. 1.
Specifically, the polynucleotide sequence shown in SEQ ID No.1 is obtained by deletion, insertion and/or substitution of 1-100 bases or addition of 1-50 bases at the 5 '-end and/or the 3' -end, and has the function of the polynucleotide sequence shown in SEQ ID No. 1.
More specifically, the polynucleotide shown in SEQ ID No.1 is a polynucleotide in which 1 to 10 bases in the polynucleotide sequence are replaced by bases with similar or similar properties.
The invention provides agapanthus dehydratus ApY with adversity stress response function and protection function 2 SK 2 The gene promoter consists of a nucleotide sequence shown as SEQ ID No.1, and the activity of the promoter is induced by drought (PEG), high salt, osmosis, low temperature stress and phytohormone ABA.
Further preferably, the promoter or the nucleotide sequence shown by SEQ ID NO.1 is subjected to substitution, deletion or addition of one or more bases and still has agapanthus hygiensis dehydratum ApY 2 SK 2 Gene promoter activity, induced by drought (PEG), high salt, osmotic, low temperature stress and phytohormone ABA.
In the present invention, the term "agapanthus dehydratum ApY 2 SK 2 Gene promoter sequence "means located at ApY on the agapanthus genome 2 SK 2 Gene Open Reading Frame (ORF) 5' -upstream has a promoter agapanthus dehydratus ApY 2 SK 2 Nucleotide sequence of activity of gene expression.
In the present invention, "isolated DNA" or "purified DNA" means that the DNA or fragment has been isolated from the sequences which flank it in the natural state, and that the DNA or fragment has been separated from the components which accompany the nucleic acid in the natural state and from the proteins which accompany it in the cell.
The promoter is obtained by cloning from separated agapanthus DNA by a chromosome walking method, and replaces a CaMV35S gene expression frame on plasmid pBI 121. Modified agapanthus dehydratus ApY 2 SK 2 The recombinant plasmid of the gene promoter sequence was designated ApY 2 SK 2 -P::GUS。
In a second aspect, the invention provides the use of the polynucleotide as a promoter element for the expression of a plurality of proteins or polypeptides in a plant bioreactor.
In particular, the plant bioreactor may be a plant host cell.
More specifically, the promoter elements can be used to direct inducible expression of a variety of proteins or polypeptides of different origin in a plant host cell.
More specifically, the polynucleotide sequence (promoter) does not promote expression of the downstream gene under normal growth conditions, but activity of the polynucleotide sequence (promoter) is stimulated to promote expression of the downstream gene under stress and/or hormone induction.
Further, the proteins or polypeptides from different sources can be proteins with any molecular weight, such as various proteins from plant sources, various proteins from animal or bacterial sources, antibodies, and the like.
In a third aspect, the invention provides the use of the polynucleotide as an expression vector for a plant bioreactor.
The polynucleotide can be used as a promoter for guiding the expression of the foreign protein to construct or modify an expression vector of a plant bioreactor.
The plant bioreactor expression vector to be modified includes but is not limited to: pCAMBIA series vectors, pBI series vectors (e.g., pBI121), and pPZP series vectors, such as pSN1301 (a transformable dicot plant), pUN1301 (a transformable monocotyledon plant), pRTL2, pRTL2-GFP, pRTL2-CFP, pRTL 2-RFP, pRTL2-YFP, and the like.
In a fourth aspect, the present invention provides a plant bioreactor expression vector comprising at least one of said polynucleotides as a promoter element, wherein said promoter directs the expression of a foreign protein. Specifically, the beta-glucuronidase gene is GUS.
Preferably, the expression vector is obtained by replacing the CaMV35S promoter in the vector pBI121 with the polynucleotide (promoter).
In a fifth aspect, the invention provides a host cell comprising said expression vector or having exogenous said polynucleotide integrated into its genome.
In a sixth aspect, the present invention provides a method for expressing a protein or polypeptide, comprising the steps of:
1) constructing recombinant expression vectors of the proteins or the polypeptides from the different sources by adopting the expression vectors;
2) transfecting the recombinant expression vector obtained in step 1 into a host, and culturing the host under conditions suitable for expressing the protein or polypeptide.
Preferably, the host may be a host cell, and in one embodiment of the present invention, the host is tobacco lamina.
Preferably, the induction is specifically performed by PEG, mannitol, NaCl, low temperature (4 ℃) and ABA under the condition of properly expressing the protein or the polypeptide.
Preferably, after step 2), the protein or polypeptide may be isolated from the culture or the plurality of different sources of protein or polypeptide may be detected by a detection instrument. In one embodiment of the invention, the protein or polypeptide is GUS.
Recombinant expression vectors for said various proteins or polypeptides of different origin can be constructed by conventional recombinant DNA techniques, for example by inserting a gene segment of interest into the multiple cloning site of said plant bioreactor cell expression vector under the control of a promoter according to the present invention. The gene segment of interest should at least comprise the entire coding region of the gene for a number of different sources of proteins or polypeptides.
The obtained host cell transferred with the recombinant expression vector can be cultured by a conventional method to express the foreign protein or polypeptide. Depending on the host cell used, the medium used in the culture may be selected from various known suitable media or culture solutions, and the culture may be carried out under conditions suitable for the growth of the host cell and the expression of the foreign protein or polypeptide. If desired, the recombinant protein can be isolated and purified by various separation methods using its physical, chemical and other properties or the addition of a purification tag. These methods are well known to those skilled in the art. Examples of such methods include, but are not limited to: treating with protein precipitant (salting out method), centrifuging, breaking cell wall by osmosis, treating with ultra-high pressure, ultracentrifuging, molecular sieve chromatography (gel filtration), adsorption chromatography, ion exchange chromatography, High Performance Liquid Chromatography (HPLC), and other liquid chromatography techniques and combinations thereof.
According to agapanthus dehydratin ApY 2 SK 2 cDNA sequence of protein gene, through genome walking method we clone to dehydrin ApY closely related to agapanthus responding to stress 2 SK 2 Promoter sequence of gene on genome. ApY will be mixed 2 SK 2 The promoter of the gene is connected with GUS, the arabidopsis thaliana is transformed by an agrobacterium-mediated method, and the response of the promoter to drought, osmosis, high salt, low temperature (4 ℃/8 ℃), high temperature (40 ℃/45 ℃) and ABA (50M/100M) is detected. The detection of GUS enzyme activity proves that the promoter has the activity of the promoter, and is enhanced by four stresses of drought, osmosis, high salt and low temperature and hormone induction activity. In order to further define the regulation function of different cis-acting elements on the promoter on stress signals, a plurality of ApY are constructed 2 SK 2 The expression vector fused with promoter fragments with different deletions in length and GUS gene is transformed into arabidopsis thaliana, and the detection result of GUS enzyme activity shows that ApY 2 SK 2 The response of the promoter to stress and hormone is mainly regulated and controlled by MBS, LTR, ABRE and other cis-acting elements. Thus, the effect of the dehydrin gene in agapanthus responding to drought, osmosis, high salt, low temperature stress and ABA is shown. In addition, compared with other promoters, the promoter provided by the invention is suitable for promoting the expression of a reporter gene such as a beta-glucuronidase gene GUS gene in a plant, and has the activity characteristic of being controllable by induction.
Drawings
Other characteristics, objects of the invention, through reading the detailed description of non-limiting embodiments with reference to the following figures
ApY in FIG. 1 2 SK 2 Bioinformatics analysis of gene promoter sequences for putative cis-regulatory element sequences, wherein the cis-regulatory elements are boxed and element names are noted;
ApY in FIG. 2 2 SK 2 Plant reactor expression vector ApY with gene promoter 2 SK 2 -P: the structural diagram of GUS;
FIG. 3 shows two plant reactor expression vectors (pBI121, ApY) 2 SK 2 GUS) change in GUS enzyme activity expression under different treatments after transformation of Arabidopsis thaliana;
ApY in FIG. 4 2 SK 2 A promoter deletion expression vector segmentation scheme;
FIG. 5 shows five plant reactor expression vectors (ApY) 2 SK 2 GUS, Sp-946 GUS, Sp-670 GUS, Sp-262 GUS and Sp-167 GUS) after Arabidopsis thaliana transformation.
Detailed Description
Example 1 African agapanthus dehydratin ApY 2 SK 2 Cloning of Gene promoters
1. Obtaining plant material
Taking agapanthus leaf tissues for extracting DNA;
extraction of DNA
Use of
Figure GDA0003698690440000051
Extracting agapanthus genome DNA by using a Plant Genomic DNA Kit, taking 100mg of agapanthus leaf tissue, adding liquid nitrogen for full grinding, and transferring the powder into a 1.5ml EP tube; adding 250 μ L RB1 solution and 15 μ L RNase solution, mixing well, incubating in 55 deg.C water bath for 15min, centrifuging, and collecting supernatant. Adding 100 μ L PB1 solution into the supernatant, mixing, standing on ice for 5min, centrifuging, and collecting supernatant; adding 375 μ L BBI solution into the supernatant, adding the mixture into a centrifugal column, and centrifuging to remove eluate; addingAdding 500 μ L CB1 solution, centrifuging, and discarding the effluent; adding 500 μ L of WB1 solution, centrifuging to remove effluent, repeating for 1 time, centrifuging at 12000 Xg for 2min, and removing WB1 solution; placing the column on a new EP tube, adding 100 μ L of preheated EB solution (60-70 deg.C) into the center of the column, and centrifuging at 12000 × g for 1min to elute DNA; repeating the steps, and performing secondary DNA elution. Storing the obtained DNA group at-20 ℃ for later use;
3. cloning of Gene promoters
According to agapanthus dehydratin ApY 2 SK 2 The primers for gene cDNA full-length sequence design were carried out by ApY using chromosome Walking method (Genomic Walking kit from Takara Clotech) 2 SK 2 Cloning gene 5' -upstream promoter;
based on ApY 2 SK 2 Coding gene sequence, first chromosome walking, designing downstream specific primer ApY 2 SK 2 SP1/2/3, cloned ApY using AP Primer from Genome Walking kit as upstream Primer 2 SK 2 The PCR program is shown in table 1, and after 3 rounds of thermal asymmetric nested PCR, the upstream promoter sequence of the 5' -end is obtained, and the fragment is recovered and sequenced; second chromosome walking at ApY 2 SK 2 4 rounds of PCR are carried out by using-SP 2-1/2/3/4 as specific primers, PCR products are recovered and sequenced, and the sequencing results are spliced by using Clustalx (1.81) software to obtain ApY with the length of about 1200bp 2 SK 2 -P sequence SEQ ID NO 1.
Primer ApY 2 SK 2 -SP1:5′-GCCCTGTTGCGGTAGTCGTAGT-3′(SEQ ID NO:2)
Primer ApY 2 SK 2 -SP2:5′-TCTTCTTCCTCCTCCCCCCCAT-3′(SEQ ID NO:3)
Primer ApY 2 SK 2 -SP3:5′-CCGTAAGCGTCCGTCTGTTGGA-3′(SEQ ID NO:4)
Primer ApY 2 SK 2 -SP2-1:5′-GAGAGCGTGGAGGTGAGAATGA-3′(SEQ ID NO:5)
Primer ApY 2 SK 2 -SP2-2:5′-AAGGGCGATTGGGATGTGGGTT-3′(SEQ ID NO:6)
Primer ApY 2 SK 2 -SP2-3:5′-CGTATGGGAAGATCATGGCAGG-3′(SEQ ID NO:7)
Primer ApY 2 SK 2 -SP2-4:5′-TGTGAGTGAAGGCCAGGGCTAA-3′(SEQ ID NO:8)
TABLE 1 thermal asymmetric PCR reaction conditions
Figure GDA0003698690440000061
ApY 2 SK 2 The complete promoter sequence of gene 5' -upstream is (SEQ ID NO:1)
Figure GDA0003698690440000062
Figure GDA0003698690440000071
Plant cis-element database plantarCARE pair ApY 2 SK 2 The promoters were subjected to bioinformatic predictive analysis. The analytical results are shown in Table 2, ApY 2 SK 2 The promoter contains structural elements which are commonly used by various promoters and cis-acting elements related to adversity stress and hormone response.
TABLE 2 ApY 2 SK 2 Cis-acting elements in gene promoters
Figure GDA0003698690440000072
Figure GDA0003698690440000081
Figure GDA0003698690440000091
Example 2 expression vector construction
Plasmid pBI121 (purchased from Youbao, Hunan, China) as a template ApY of example 1 2 SK 2 After the gene promoter sequence is cut by HindIII and Xbal, the gene promoter sequence is connected between HindIII and Xbal cutting sites of pBI121 to replace the original CaMV35S promoter, and ApY is completed 2 SK 2 And (3) establishing an expression frame for expressing a GUS gene in a gene promoter sequence. The obtained recombinant plasmid was designated ApY 2 SK 2 GUS (FIG. 2).
Example 3 analysis of promoter expression Activity under different stress and exogenous hormone treatment
To study the dehydrin gene ApY 2 SK 2 The pBI121 empty vector (35S promoter) and ApY were used as response mechanisms under stress and hormone treatment, respectively 2 SK 2 GUS is transformed into agrobacterium GV3101 by a freeze-thaw method, and the obtained positive monoclonal is used for transforming arabidopsis by using a flower dipping method. Transgenic Arabidopsis thaliana was treated with PEG8000 (20%), 400mM mannitol, 3% NaCl, low temperature (8 ℃/4 ℃), high temperature (40 ℃/45 ℃) and ABA (100. mu.M), respectively, with no treatment as a blank control and a positive control containing a 35S strong promoter. The specific treatment method comprises the following steps: disinfecting T3 generation Arabidopsis seeds, sowing the seeds on an MS culture medium, and respectively transferring the seeds after 14 days into an MS solid culture medium containing 20% PEG, 400mM mannitol and 200 mM NaCl to carry out drought, osmotic stress and salt stress; transferring the mixture into an environment with the temperature of 8 ℃/4 ℃ for low-temperature stress; transferring the mixture into an environment of 40 ℃/45 ℃ for high-temperature stress; spraying 100 mu M abscisic acid solution for hormone treatment, and carrying out quantitative detection on GUS enzyme activity on the obtained sample.
The results show that: under normal growth conditions, ApY 2 SK 2 Full-length promoter (ApY) 2 SK 2 -P) has certain expression activity (110nM MU/min mg) -1 ) (ii) a Under different stresses and hormone signal treatment, the GUS enzyme activity of the positive control has no obvious change (about 742nM MU/min. mg) -1 ) And ApY 2 SK 2 There was a significant difference in the GUS protein activity driven by P (FIG. 3). ApY 2 SK 2 The response of P to drought and low temperature stress was most significant (GUS enzyme activity was 6.9 and 8.4 times higher than the control, respectively); for osmotic, salt stress and ABA treatmentThe positive response also indicates that the successfully constructed promoter expression vector has expression activity and the promoter activity can be greatly up-regulated under the induction of various stresses and hormones (figure 3, CK in the figure represents blank control, namely, the arabidopsis thaliana is not treated at all).
Example 4 analysis of promoter deletion response to stress and hormone treatment
ApY is selected according to the distribution of stress signal, hormone signal response element and endosperm development element 2 SK 2 The promoter was subjected to 5' -end deletion segmentation and cloning to construct a binary expression vector of 5 promoters and GUS reporter genes (FIG. 4, only the action elements of interest are listed). A promoter-deleted expression vector was constructed with reference to example 2, and the obtained recombinant plasmid was designated Sp-1199 (ApY) 2 SK 2 -P), Sp-946, Sp-670, Sp-262 and Sp-167; reference example 3 Arabidopsis thaliana was transformed with the deletion expression vector, and GUS enzyme activity was measured after stress and hormone treatment.
The results show (FIG. 5) that under different stresses and hormone treatments, GUS enzyme activity showed a decreasing trend as the length of the promoter fragment decreased. Under the treatment of stress hormone, compared with a control group, the GUS enzyme activity of each deletion fragment of the promoter is obviously up-regulated. Under the treatment of PEG, GUS enzyme activities of Yp-1199 and Yp946 fragments are respectively and obviously up-regulated by 6.9 and 1.8 times, and the-1199-670 fragments contain MBS cis-acting elements and are important regions for responding drought. Under low-temperature treatment, the GUS enzyme activity of the Yp-1199 fragment is up-regulated by 8.4 times, and compared with a control, the GUS enzyme activities of Sp-946, Sp-670, Sp-262 and Sp-167 have no significant difference, and the-1199-946 segment has a low-temperature stress response element LTR which is an important regulation and control region for low-temperature response. Under NaCl and mannitol treatment, GUS enzyme activities of Yp-1199 and Yp-946 fragments are significantly different from those of a control, and the-1199-670 fragment is a main segment responding to NaCl and mannitol. Under ABA hormone treatment, GUS enzyme activities of Yp-1199, Yp946 and Yp-670 fragments are respectively up-regulated by 4.3, 5.6 and 3.2 times, while the-400-262 area contains 4 tandem ABRE response elements ABRE, the-262-167 contains 1 ABRE, and the experimental result shows that the 4 tandem ABRE of the previous section are very important for the response of exogenous ABA hormone signals. In conclusion, ApY 2 SK 2 The promoter can respond to various stress and ABA hormones, wherein the response to drought, low-temperature stress and ABA hormones is the most remarkable. -1199- "262 is the main segment of stress and hormone response, which contains a number of homeopathic regulatory elements related to stress and hormones, such as MBS, LTR and ABRE. ABRE differs from MBS and LTR elements at ApY 2 SK 2 Multiple tandem is required in a promoter to respond to ABA hormone.
In conclusion, the present invention effectively overcomes various disadvantages of the prior art and has high industrial utilization value.
It should be noted that the prior art in the protection scope of the present invention is not limited to the examples given in the present application, and all the prior art which is not inconsistent with the technical scheme of the present invention, including but not limited to the prior patent documents, the prior publications and the like, can be included in the protection scope of the present invention.
In addition, the combination of the features in the present application is not limited to the combination described in the claims of the present application or the combination described in the embodiments, and all the features described in the present application may be freely combined or combined in any manner unless contradictory to each other.
The foregoing description of specific embodiments of the present invention has been presented. It is to be understood that the present invention is not limited to the specific embodiments described above, and that various changes or modifications may be made by one skilled in the art within the scope of the appended claims without departing from the spirit of the invention. The embodiments and features of the embodiments of the present application may be combined with each other arbitrarily without conflict.
Sequence listing
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<141> 2021-03-12
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aaacaaacag aataacaagg acaagaacag aaaaaaataa aataaagaga aaagagagaa 480
aataagacat accaaatcta cttaaataaa cctacactac ataaactagt gcatccgaat 540
tctattttta cgtatcctct ttctagtaat ctagtaatct cacgttgaag ttgatcaaga 600
ctgttataac tagtccgatg ttcgttatat ggaattcaaa caagaacaac agttttcctt 660
agtttgtaat gaaattttat ttttctaatt tggagaaatt tttatatttg tttattttat 720
tttcttcttt ttcccccgct tttggggtaa tttctcctta attagccctg gccttcactc 780
acatttctcc ccacaacaat taacactctc ccacattacc aggtaccacg tacgtcccct 840
cacatgcatg catcaacacg tgggctcgct ccctgccatg atcttcccat acgtggacac 900
cccacacgtg ccaccctccc cttaacccac atcccaatcg cccttatcct caaacgccac 960
cgcccacacg tgtcccctct ctgtccccga tgcgcaaacg tcgcatgcca ccccttcaca 1020
cgcgtcgctc cccccaaatg aaatctctca ttctcacctc cacgctctca cgtaccctct 1080
atataaactc accaccttct cctcatctct catcacaaaa tcgaaagatc agttgaaaaa 1140
agagagagga aaagaacaag aaaaccattt cgagagaatt tgatcagttt tggagagcga 1200
tg 1202
<210> 2
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 2
gccctgttgc ggtagtcgta gt 22
<210> 3
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 3
tcttcttcct cctccccccc at 22
<210> 4
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 4
ccgtaagcgt ccgtctgttg ga 22
<210> 5
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 5
gagagcgtgg aggtgagaat ga 22
<210> 6
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 6
aagggcgatt gggatgtggg tt 22
<210> 7
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 7
cgtatgggaa gatcatggca gg 22
<210> 8
<211> 22
<212> DNA
<213> Artificial Sequence
<400> 8
tgtgagtgaa ggccagggct aa 22

Claims (5)

1. African agapanthus dehydratin ApY 2 SK 2 Use of a gene promoter sequence, wherein the agapanthus dehydratum ApY 2 SK 2 The gene promoter sequence is used as a promoter element for expression of various proteins or polypeptides in a plant bioreactor;
the agapanthus dehydratum ApY 2 SK 2 The gene promoter sequence is a polynucleotide sequence shown as SEQ ID number 1.
2. The use of claim 1, wherein said promoter element is used to direct the inducible expression of a plurality of proteins or polypeptides of different origin in a plant bioreactor.
3. A plant bioreactor expression vector comprising the polynucleotide sequence of claim 1 as a promoter element, wherein expression of a foreign protein is directed by said promoter element.
4. The plant bioreactor expression vector of claim 3, wherein the expression vector is obtained by replacing the CaMV35S promoter in the vector pBI121 with a polynucleotide sequence shown in SEQ ID No. 1.
5. A method for expressing a protein or polypeptide, comprising the steps of:
step S1: constructing recombinant expression vectors of a plurality of proteins or polypeptides of different origins by using the expression vectors of claim 3 or 4;
step S2: transfecting the recombinant expression vector obtained in step S1 into a host, and culturing the host under conditions suitable for expression of the protein or polypeptide.
CN202110350763.9A 2021-03-31 2021-03-31 African agapanthus dehydratin ApY 2 SK 2 Gene promoter sequence and use thereof Expired - Fee Related CN113151268B (en)

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