CN111440814A - Insect-resistant fusion gene mCry1AbVip3A, expression vector and application thereof - Google Patents

Abstract

The invention discloses an insect-resistant fusion gene mCry1AbVip3A, wherein the nucleotide sequence of the gene mCry1AbVip3A is shown as (a), (b) or (c): (a) SEQ ID NO: 1; or (b), encodes SEQ ID NO: 2; or (c), and SEQ ID NO: 1 under stringent hybridization conditions, and the protein encoded by the nucleotide has the function of the transcription factor of mCry1AbVip 3A. In vitro toxicity test proves that the modified and synthesized fusion gene toxigenin has obvious insecticidal effect on corn borer and spodoptera littoralis, can be stably and efficiently expressed in monocotyledons, and is further used for producing insect-resistant transgenic plants.

Description

Anti-insect fusion gene mCry1AbVip3A, its expression vector and application

technical field

The present invention relates to the field of genetic engineering and biological control, in particular, to an anti-insect fusion gene, its expression vector and application

Background technique

Pests cost about $8 billion a year in global agricultural production. At present, the control of pests mainly relies on the use of chemical pesticides, but pesticide residues can cause harm to human health. Therefore, people have successfully selected transgenic crops that can produce insecticidal toxins for insect resistance. At present, transgenic insect-resistant corn and cotton have been planted in large quantities.

Substances with strong insecticidal activity against pests such as Agrotis ipsilon, Spodoptera exigua and Spodoptera frugiperda were found in the fermentation supernatant of strain Bt AB88. It is expressed from the mid-phase to the spore stage, and is named vegetative insecticidal proteins (Vip) (Estruch, 1996). The Bt gene from Bacillus thuringHansis can also encode insecticidal protein crystals, which produce delta-endotoxin parasporal crystal proteins during spore formation, and these proteins have high insecticidal activity. The principle of action is that the anti-insect protein can be dissolved by alkaline intestinal fluid and hydrolyzed into smaller active toxin fragments - core fragments (Hofte and Whiteley, 1989). The active fragment is resistant to further hydrolysis by proteases, and the activated protein binds to brush-like vesicles in the insect gut, causing perforation to affect osmotic balance, cell swelling and lysis, and target organisms stopping feeding and eventually dying. Studies have shown that the intestinal epithelial cells of many target pests have high-affinity binding sites for Bt proteins (Hofte and Whiteley, 1989). Compared with Cry-like proteins, the research on the insecticidal mechanism of Vip3-like proteins has just started. The initial mechanism study found that epithelial columnar cells swelled into spherical shape after feeding Vip3A toxin for 24 hours, and the shape of goblet cells changed slightly. , the cell damage was not obvious at this stage; after 48h, the columnar cells continued to deteriorate, the intestinal lumen was filled with fragments of collapsed cells, and the goblet cells were significantly damaged, but they were still connected to the basement membrane; after 72h, the epithelial layer was completely detached, and the insect died, but No obvious cellular damage was seen in the foregut and hindgut. Symptoms triggered by ingestion of Vip3A by susceptible insects were similar to delta-endotoxin, but delayed in time. Meanwhile, in vivo immunolocalization assay showed that Vip3A could specifically bind to the top microvilli, and most of them were concentrated in columnar cells, and no signal was detected in goblet cells. The columnar cells in the midgut tissue mainly perform the functions of secreting digestive enzymes and absorbing digestive products. After Vip3A is activated by the protease in the intestinal juice, in the process of being absorbed by the columnar cells, it may bind to some receptor proteins on the cell membrane to cause the cell membrane Perforation, or triggering a certain signaling pathway, eventually worm death (Yu CG and Mullins MA et al., 1997).

In recent years, there have been many studies on the insecticidal protein Vip in the vegetative period. The Vip-like proteins that have been discovered are mainly divided into four categories, namely Vip1, Vip2, Vip3 and Vip4. Vip1/Vip2 constitute a binary toxin, which mainly acts on Coleoptera and Hemiptera, and there are relatively few studies (Ellis RT and Stamp L, 2002). Huazhong Agricultural University discovered a vip4 gene, but there are no related reports on its insecticidal mode of action and target specificity.

Saraswathy et al. constructed a chimeric protein of Vip3Aa14 and Cry1Ac. The chimeric protein mainly exists in the form of inclusion bodies and has good stability to trypsin, but only maintains the insecticidal activity of Cry1Ac and does not show synergistic effect. In addition, the C-terminus of Cry1C can significantly enhance the synthesis of Vip3Aa7 and help it to form inclusion bodies in BMB171, but its biological activity is significantly lower than that of the protoxin Vip3Aa7 (Song R et al., 2008). The reduced virulence of the above fusion proteins may be caused by the fusion protein not folding properly. However, the fusion protein constructed by Dong et al. fused Vip3Aa7 with the N-terminus of Cry9Ca, and the synergistic effect factor of Plutella xylostella reached 4.79, which was much higher than the synergistic effect of mixing the two protoxins. Yu et al. co-expressed Vip3Aa29 and Cyt2Aa3, and the co-expressed proteins had a synergistic effect on Spodoptera exigua and rice borer. Fusion or co-expression of Vip3 with insecticidal toxins from different sources can effectively enhance its virulence to target insects, expand the insecticidal spectrum, and delay the emergence of insect resistance. In addition, Vip3 can be integrated into the chromosome of insect virus, which can not only effectively spread Vip3A, but also improve the insecticidal virulence of the virus.

Compared with the single Cry1Ab or VIP3A gene, the anti-insect fusion gene of the present invention can resist both corn borer and Spodoptera frugiperda.

SUMMARY OF THE INVENTION

The purpose of the present invention is to provide an anti-insect fusion gene and its expression vector which can be stably and efficiently expressed in plants.

Another object of the present invention is to provide the application of the insect-resistant fusion gene in improving the insect resistance of transgenic plants.

The object of the present invention can be further realized by adopting the following technical measures:

Under the condition that the amino acid composition of the translation protein of the sequence Vip3A remains unchanged, base substitutions are performed with the codons preferred by monocotyledonous plants to initially obtain a modified DNA sequence; Contains AT sequence and commonly used restriction enzyme cleavage site (SacI), and then corrects and eliminates by replacing codons; then removes the terminator from Cry1Ab-Ma sequence and places it on the N-terminus of synthetic Vip3A; confirms and chemically synthesizes the modified The nucleotide sequence of the fusion gene mCry1AbVip3A is shown in SEQ ID No. 1; according to the needs of cloning, restriction endonuclease recognition site sequences are added at both ends of the sequence and constructed into a corresponding expression vector.

The fusion gene of the present invention is operably linked to the prokaryotic expression vector, and the toxicity of the synthetic Bt gene expression product of the present invention to corn borer and Spodoptera frugiperda can be rapidly and preliminarily detected. The fusion gene of the present invention is operably linked to a plant expression vector, and the expression vector is introduced into the corresponding Agrobacterium, and then genetically transformed by the Agrobacterium-mediated method to cultivate insect-resistant transgenic maize. Other crops or fruit trees can also be genetically transformed to have anti-insect activity.

Those skilled in the art can also transform the gene of the present invention into bacteria or fungi, produce the fusion anti-insect protein through large-scale fermentation, and prepare it into an insecticide for the control of crop pests.

In order to achieve the object of the present invention, the present invention first provides an anti-insect fusion gene mCry1AbVip3A, the nucleotide sequence of the gene mCry1AbVip3A is shown in (a), (b), or (c):

(a), the nucleotide shown in SEQ ID NO: 1; or

(b), a nucleotide encoding the amino acid shown in SEQ ID NO: 2; or

(c), a nucleotide capable of hybridizing with the complementary sequence of SEQ ID NO: 1 under stringent hybridization conditions, and the protein encoded by the nucleotide has the function of a mCry1AbVip3A transcription factor.

Further, the present invention also provides that the amino acid sequence encoded by the anti-insect fusion gene mCry1AbVip3A is shown in SEQ ID NO: 2.

Further, the present invention also provides a vector containing an anti-insect fusion gene, preferably a bivalent vector.

Further, the present invention also provides a host cell containing the above-mentioned vector, preferably EHA105.

Further, the present invention also provides transformed plant cells containing the anti-insect fusion gene.

Furthermore, the present invention also provides the application of the insect-resistant fusion gene in improving the insect resistance of transgenic plants. Preferably, the plants are crops, fruit trees or vegetables, such as corn, rice, potato, cotton and the like.

The present invention has at least the following advantages and beneficial effects:

Compared with Cry1Ab-Ma and Vip3A alone, the insecticidal fusion protein of the invention can be compatible with the corn borer resistance of Cry1Ab-Ma and the Spodoptera frugiperda resistance of Vip3A.

After the insect-resistant fusion gene is introduced into maize, a stably inherited transformant can be obtained. In addition, the gene can also transform cotton, rice, vegetables and other crops to have corresponding anti-insect activity, thereby reducing the use of pesticides and reducing environmental pollution, which has important economic value and broad application prospects.

Description of drawings

Figure 1 Map of plant expression vector containing mCry1AbVip3A gene fragment.

Figure 2. Detection results of transgenic maize mCry1AbVip3A colloidal gold test paper. In the figure, the color band on the test strip is the quality control line, and the bottom is the target protein detection line; the left 1 is the negative control, and the others are the transgenic maize samples.

Detailed ways

The following examples are intended to illustrate the present invention, but not to limit the scope of the present invention. Unless otherwise specified, the technical means used in the examples are conventional means well known to those skilled in the art.

Example 1 Synthesis of anti-insect fusion gene

According to the original nucleotide sequence of the Vip3A gene (gene ID DQ539888.1), while keeping the overall amino acid composition of the protein unchanged, the bases were replaced with the codons preferred by monocotyledonous plants, and a modified DNA sequence was initially obtained. ; Exclude AT-rich sequences (such as ATTTA, AATGAA, etc.) and commonly used restriction enzyme cleavage sites (SacI) that cause plant transcription instability in the DNA sequence, and then correct and eliminate them by replacing codons; and in 5 Add Cry1Ab-Ma sequence to the ' end; determine and chemically synthesize the modified anti-insect fusion gene mCry1AbVip3A, whose nucleotide sequence is shown in SEQID No.1; add restriction endonuclease recognition sites at both ends of the sequence according to cloning needs dot sequence and construct into the corresponding expression vector.

Example 2 Expression of anti-insect fusion gene in prokaryotic system and toxicity detection of product

In order to detect the in vitro expression of the modified anti-insect gene and its toxicity to corn borer, we constructed a prokaryotic expression vector pET-mCry1AbVip3A.

Design primer sequences (upstream primer F1: as shown in SEQ ID No. 3, 5'-GACGACGACAAGGCCATGGATGGACAACAACCCG-3'; F2: as shown in SEQ ID No. 4, 5'-ACGACGACAAGGCCATGGATGAACAAGAACAAC-3'; downstream primer R1: 5' -GGTGGTGGTGGTGCTCGAGCTTGATGCTCACGTC-3' as shown in SEQ ID No. 5; R2: as shown in SEQ ID No. 6, 5'-GGTGGTGGTGGTGCTCGAGGTACTCCGCCTCGAA-3').

Using the constructed pTF101.1-mCry1AbVip3A plant expression plasmid (Figure 1) as a template, and using F1 and R1 as primers, amplify the anti-insect fusion gene, gel recovery kit (Tiangen Biochemical Technology (Beijing) Co., Ltd., DP209 ) recovery and purification of the above insect-resistant gene fragments. The pET30a+ was digested with restriction enzymes NcoI and XhoI, and the 4.2kb fragment was recovered and purified with a gel recovery kit. The recovered fragment was ligated with the digested pET30a+ fragment, and the resulting prokaryotic expression plasmid was constructed and named pET-mCry1AbVip3A. Digestion with different restriction enzymes indicated that the vector was constructed correctly.

The constructed prokaryotic vector pET-mCry1AbVip3A was used to transform BL21(DE3) competent cells. Bt protein was extracted from infected Escherichia coli BL21 bacteria after induction by IPTG, and the insect test was carried out with corn borer and Spodoptera frugiperda. Specific steps are as follows:

①Inoculate a single colony of Escherichia coli BL21 in 5ml LB (containing kanamycin) liquid medium, and cultivate overnight at 37°C.

②Inoculate the bacterial liquid into 500ml LB (containing kanamycin) liquid medium, and cultivate to OD≈0.5.

③Add IPTG to the final concentration of 0.5mM and continue to shake for 4 hours.

④ Centrifuge at 4000 rpm for 10 min to collect bacterial cells.

⑤ Add 36ml of lysis buffer (2mM Tris-HCl; 0.2mM CaCl ₂ ; pH=8.0), resuspend the cells, add lysozyme to a final concentration of 1mg/ml, and place on ice for 30min.

⑥Ultrasonic break the cells (breaking parameters: work 10s, interval 5s, 40 times, ice bath breaking), centrifuge at 4000rpm for 10min, and collect the supernatant.

⑦ The collected supernatant was added to the feed to feed corn borer and Spodoptera frugiperda.

Toxicity determination of corn borer: Weigh 5g of Asian corn borer artificial feed, put it in a petri dish, cut the feed into 1-2mm slices, apply 1ml of protein liquid (50ng/ul) evenly on the surface of the feed, and infiltrate it for 2h . After 2 hours, 10 corn borer larvae hatched for 24 hours were inserted into each petri dish. The experiments were performed with the same volume of distilled water as a control. 5 replicates were set for each treatment. Placed in an artificial climate incubator with a temperature of 28°C, a photoperiod of 16h:8h (L:D), and a relative humidity of 80%. The number of surviving larvae was surveyed and recorded every other day.

Toxicity determination of Spodoptera frugiperda: Weigh 5g of artificial feed, put it in a worm box, cut the feed into 1-2mm slices, apply 1ml of protein liquid (50ng/ul) evenly on the surface of the feed, and infiltrate it for 2h . After 2 hours, one Spodoptera frugiperda larvae hatched for 24 hours was inserted into each insect box. The experiments were performed with the same volume of distilled water as a control. 30 replicates were set up for each treatment. Placed in an artificial climate incubator with a temperature of 26°C, a photoperiod of 16h:8h (L:D), and a relative humidity of 60%. The number of surviving larvae was surveyed and recorded daily.

Data analysis: excel 2007 was used to preliminarily organize the data, and the survival rates of corn borer and Spodoptera frugiperda larvae at different feeding times were calculated; spss 23 was used to conduct variance analysis and significant difference analysis on the corn borer survey data. The results showed that on the 6th day, no larvae of corn borer and Spodoptera frugiperda were fed with mCry1AbVip3A protein (Table 1, Table 2). The test results showed that mCry1AbVip3A had strong insecticidal activity.

Table 1: Toxicity to corn borer larvae

Table 2: Toxicity to Spodoptera frugiperda larvae

Example 3 Expression of mCry1AbVip3A gene in transgenic maize

The pTF101.1-mCry1AbVip3A (Fig. 1) insert sequence was introduced into the callus of recipient maize by Agrobacterium-mediated transformation (strain EHA105), and the transgenic maize was obtained after screening with the herbicide bialaphos. Detection of target protein mCry1AbVip3A (Bt-Cry1Ab/1Ac colloidal gold immunological detection):

①Take about 1cm ² of fresh young leaves and put them in a 1.5ml Eppendorf tube, add 500ul SEB4 sample extraction buffer to the tube and grind it with a grinder.

②Take out the test strip (Beijing Gennis Biotechnology Co., Ltd., item number: STX06200/0050), hold the top of the test strip, and insert the marked end into the centrifuge tube. The control line will appear within 3-5 minutes, and if the sample is positive, the test line will appear.

The expression of mCry1AbVip3A in leaves of 6 transgenic plants was determined by colloidal gold strip detection method. The results showed that mCry1AbVip3A protein was expressed in transgenic maize leaf tissue (Figure 2).

Although the present invention has been described in detail above with general description and specific embodiments, some modifications or improvements can be made on the basis of the present invention, which will be obvious to those skilled in the art. Therefore, these modifications or improvements made without departing from the spirit of the present invention fall within the scope of the claimed protection of the present invention.

sequence listing

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

<120> Anti-insect fusion gene mCry1AbVip3A, its expression vector and application

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Ile Met Ala Ser Pro Val Gly Phe Ser Gly Pro Glu Phe Thr Phe Pro

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Asn Leu Gly Ser Gly Thr Ser Val Val Lys Gly Pro Gly Phe Thr Gly

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Tyr Ala Ser Thr Thr Asn Leu Gln Phe His Thr Ser Ile Asp Gly Arg

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Pro Ile Asn Gln Gly Asn Phe Ser Ala Thr Met Ser Ser Gly Ser Asn

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Leu Gln Ser Gly Ser Phe Arg Thr Val Gly Phe Thr Thr Pro Phe Asn

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Phe Ser Asn Gly Ser Ser Val Phe Thr Leu Ser Ala His Val Phe Asn

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Thr Arg Ala Leu Pro Ser Phe Ile Asp Tyr Phe Asn Gly Ile Tyr Gly

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Phe Ala Thr Gly Ile Lys Asp Ile Met Asn Met Ile Phe Lys Thr Asp

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Thr Gly Gly Asp Leu Thr Leu Asp Glu Ile Leu Lys Asn Gln Gln Leu

660 665 670

Leu Asn Asp Ile Ser Gly Lys Leu Asp Gly Val Asn Gly Ser Leu Asn

675 680 685

Asp Leu Ile Ala Gln Gly Asn Leu Asn Thr Glu Leu Ser Lys Glu Ile

690 695 700

Leu Lys Ile Ala Asn Glu Gln Asn Gln Val Leu Asn Asp Val Asn Asn

705 710 715 720

Lys Leu Asp Ala Ile Asn Thr Met Leu Arg Val Tyr Leu Pro Lys Ile

725 730 735

Thr Ser Met Leu Ser Asp Val Ile Lys Gln Asn Tyr Ala Leu Ser Leu

740 745 750

Gln Ile Glu Tyr Leu Ser Lys Gln Leu Gln Glu Ile Ser Asp Lys Leu

755 760 765

Asp Ile Ile Asn Val Asn Val Leu Ile Asn Ser Thr Leu Thr Glu Ile

770 775 780

Thr Pro Ala Tyr Gln Arg Ile Lys Tyr Val Asn Glu Lys Phe Glu Glu

785 790 795 800

Leu Thr Phe Ala Thr Glu Thr Ser Ser Lys Val Lys Lys Asp Gly Ser

805 810 815

Pro Ala Asp Ile Leu Asp Glu Leu Thr Glu Leu Thr Glu Leu Ala Lys

820 825 830

Ser Val Thr Lys Asn Asp Val Asp Gly Phe Glu Phe Tyr Leu Asn Thr

835 840 845

Phe His Asp Val Met Val Gly Asn Asn Leu Phe Gly Arg Ser Ala Leu

850 855 860

Lys Thr Ala Ser Glu Leu Ile Thr Lys Glu Asn Val Lys Thr Ser Gly

865 870 875 880

Ser Glu Val Gly Asn Val Tyr Asn Phe Leu Ile Val Leu Thr Ala Leu

885 890 895

Gln Ala Gln Ala Phe Leu Thr Leu Thr Thr Cys Arg Lys Leu Leu Gly

900 905 910

Leu Ala Asp Ile Asp Tyr Thr Ser Ile Met Asn Glu His Leu Asn Lys

915 920 925

Glu Lys Glu Glu Phe Arg Val Asn Ile Leu Pro Thr Leu Ser Asn Thr

930 935 940

Phe Ser Asn Pro Asn Tyr Ala Lys Val Lys Gly Ser Asp Glu Asp Ala

945 950 955 960

Lys Met Ile Val Glu Ala Lys Pro Gly His Ala Leu Ile Gly Phe Glu

965 970 975

Ile Ser Asn Asp Ser Ile Thr Val Leu Lys Val Tyr Glu Ala Lys Leu

980 985 990

Lys Gln Asn Tyr Gln Val Asp Lys Asp Ser Leu Ser Glu Val Ile Tyr

995 1000 1005

Gly Asp Met Asp Lys Leu Leu Cys Pro Asp Gln Ser Glu Gln Ile Tyr

1010 1015 1020

Tyr Thr Asn Asn Ile Val Phe Pro Asn Glu Tyr Val Ile Thr Lys Ile

1025 1030 1035 1040

Asp Phe Thr Lys Lys Met Lys Thr Leu Arg Tyr Glu Val Thr Ala Asn

1045 1050 1055

Phe Tyr Asp Ser Ser Thr Gly Glu Ile Asp Leu Asn Lys Lys Lys Val

1060 1065 1070

Glu Ser Ser Glu Ala Glu Tyr Arg Thr Leu Ser Ala Asn Asp Asp Gly

1075 1080 1085

Val Tyr Met Pro Leu Gly Val Ile Ser Glu Thr Phe Leu Thr Pro Ile

1090 1095 1100

Asn Gly Phe Gly Leu Gln Ala Asp Glu Asn Ser Arg Leu Ile Thr Leu

1105 1110 1115 1120

Thr Cys Lys Ser Tyr Leu Arg Glu Leu Leu Leu Ala Thr Asp Leu Ser

1125 1130 1135

Asn Lys Glu Thr Lys Leu Ile Val Pro Ser Gly Phe Ile Ser Asn

1140 1145 1150

Ile Val Glu Asn Gly Ser Ile Glu Glu Asp Asn Leu Glu Pro Trp Lys

1155 1160 1165

Ala Asn Asn Lys Asn Ala Tyr Val Asp His Thr Gly Gly Val Asn Gly

1170 1175 1180

Thr Lys Ala Leu Tyr Val His Lys Asp Gly Gly Ile Ser Gln Phe Ile

1185 1190 1195 1200

Gly Asp Lys Leu Lys Pro Lys Thr Glu Tyr Val Ile Gln Tyr Thr Val

1205 1210 1215

Lys Gly Lys Pro Ser Ile His Leu Lys Asp Glu Asn Thr Gly Tyr Ile

1220 1225 1230

His Tyr Glu Asp Thr Asn Asn Asn Leu Glu Asp Tyr Gln Thr Ile Asn

1235 1240 1245

Lys Arg Phe Thr Thr Gly Thr Asp Leu Lys Gly Val Tyr Leu Ile Leu

1250 1255 1260

Lys Ser Gln Asn Gly Asp Glu Ala Trp Gly Asp Asn Phe Ile Ile Leu

1265 1270 1275 1280

Glu Ile Ser Pro Ser Glu Lys Leu Leu Ser Pro Glu Leu Ile Asn Thr

1285 1290 1295

Asn Asn Trp Thr Ser Thr Gly Ser Thr Asn Ile Ser Gly Asn Thr Leu

1300 1305 1310

Thr Leu Tyr Gln Gly Gly Arg Gly Ile Leu Lys Gln Asn Leu Gln Leu

1315 1320 1325

Asp Ser Phe Ser Thr Tyr Arg Val Tyr Phe Ser Val Ser Gly Asp Ala

1330 1335 1340

Asn Val Arg Ile Arg Asn Ser Arg Glu Val Leu Phe Glu Lys Arg Tyr

1345 1350 1355 1360

Met Ser Gly Ala Lys Asp Val Ser Glu Met Phe Thr Thr Lys Phe Glu

1365 1370 1375

Lys Asp Asn Phe Tyr Ile Glu Leu Ser Gln Gly Asn Asn Leu Tyr Gly

1380 1385 1390

Gly Pro Ile Val His Phe Tyr Asp Val Ser Ile Lys

1395 1400

<210> 3

<211> 34

<212> DNA

<213> Artificial sequence

<400> 3

gacgacgaca aggccatgga tggacaacaa cccg 34

<210> 4

<211> 33

<212> DNA

<213> Artificial sequence

<400> 4

acgacgacaa ggccatggat gaacaagaac aac 33

<210> 5

<211> 34

<212> DNA

<213> Artificial sequence

<400> 5

ggtggtggtg gtgctcgagc ttgatgctca cgtc 34

<210> 6

<211> 34

<212> DNA

<213> Artificial sequence

<400> 6

ggtggtggtg gtgctcgagg tactccgcct cgaa 34

Claims (10)

1. an anti-insect fusion gene mCry1AbVip3A, is characterized in that, the nucleotide sequence of described gene mCry1AbVip3A is as shown in (a), (b) or (c): (a), the nucleotide shown in SEQ ID NO: 1; or (b), a nucleotide encoding the amino acid shown in SEQ ID NO: 2; or (c), a nucleotide capable of hybridizing with the complementary sequence of SEQ ID NO: 1 under stringent hybridization conditions, and the protein encoded by the nucleotide has the function of a mCry1AbVip3A transcription factor. 2 . The insect-resistant fusion gene mCry1AbVip3A according to claim 1 , wherein the amino acid sequence encoded by the insect-resistant fusion gene mCry1AbVip3A is shown in SEQ ID NO: 2. 3 . 3. A vector containing the gene of claim 1. 4. The carrier of claim 3, which is a bivalent carrier. 5. A host cell comprising the vector of claim 4. 6. The host cell of claim 5, which is EHA105. 7. A transformed plant cell comprising the gene of claim 1. 8. The application of the gene of claim 1 in improving the insect resistance of transgenic plants. 9. The application according to claim 8, wherein the plants are crops, fruit trees or vegetables. 10. The application according to claim 9, wherein the plant is corn, rice, potato, cotton.

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