Bombyx mori BmIMP gene and application thereof

Abstract

The invention provides a bombyx mori BmIMP gene and application thereof, and belongs to the technical field of biology and genetic engineering. The bombyx mori BmIMP gene capable of inhibiting BmNPV from infecting BmN cells is found through research, the CDS sequence of the bombyx mori BmIMP gene is shown as SEQ ID NO.1, and the amino acid sequence of protein coded by the bombyx mori BmIMP gene is shown as SEQ ID NO.2; the bombyx mori BmIMP gene can be used for preparing bombyx mori virus prevention and treatment drugs, and can be used for preparing a transformant for resisting bombyx mori nuclear polyhedrosis virus BmNPV, so that a new theoretical basis is provided for prevention and treatment of bombyx mori pyosis, and the bombyx mori BmIMP gene has good practicability.

Description

Technical Field

[0001] The invention belongs to the technical field of biology and gene engineering, and specifically relates to a silkworm BmIMP gene and an application thereof. Background Art

[0002] The silkworm, Bombyx mori, is an insect of the order Lepidoptera with important economic value. Research on the silkworm can not only provide a certain theoretical basis for other insects or mammals, but also guide the sericulture industry in a new direction. Bombyx mori nucleopolyhedrovirus (BmNPV) is a pathogen that seriously harms the silkworm production. It enters the digestive tract after oral infection. After the silkworm is infected, it will cause blood-type purulent disease, and then the silkworm will die of suppuration. The reaction is rapid and intense, causing huge economic losses to the silkworm industry around the world. Bombyx mori nucleopolyhedrovirus is highly contagious and difficult to control. It is one of the primary diseases in silkworm breeding. At present, there is still no effective method and drug to control the disease.

[0003] Since BmNPV is a serious threat to silkworm production, researchers have been hoping to find the key genes that affect silkworm resistance to the virus, and then use molecular biotechnology to improve silkworm resistance to the virus. Cloning and identifying the full-length sequence of the key genes that affect silkworm virus resistance has important theoretical value and practical significance for clarifying the mechanism of silkworm resistance to viruses and breeding resistant varieties for use in silkworm production. Summary of the invention

[0004] In view of some deficiencies in the prior art, the present invention provides a silkworm BmIMP gene and an application thereof; through research, the present invention discovers a silkworm BmIMP gene that can inhibit BmNPV from infecting BmN cells, the CDS sequence of the silkworm BmIMP gene is shown in SEQ ID NO.1, and the amino acid sequence of the protein encoded by it is shown in SEQ ID NO.2; the silkworm BmIMP gene can be used to prepare silkworm virus control drugs, and the silkworm BmIMP gene can prepare transformants resistant to silkworm nuclear polyhedrosis virus BmNPV, providing a new theoretical basis for the prevention and treatment of silkworm pus disease, and has good practicality.

[0005] In order to achieve the above technical objectives, the present invention adopts the following technical means:

[0006] The present invention first provides a silkworm BmIMP gene capable of regulating BmNPV infection of BmN cells, wherein the nucleotide sequence of the coding region (coding sequence, CDS) of the silkworm BmIMP gene is shown in SEQ ID NO. 1. Preferably, overexpression of the silkworm BmIMP gene can promote the proliferation of BmNPV in BmN cells, and inhibition / silencing of expression of the silkworm BmIMP gene can inhibit the infection ability of BmNPV in silkworm cells and silkworm bodies.

[0007] Preferably, the amino acid sequence of the protein encoded by the silkworm BmIMP gene is shown as SEQ ID NO.2.

[0008] The present invention also provides a recombinant plasmid, which contains the nucleotide sequence of the coding region of the silkworm BmIMP gene.

[0009] The present invention also provides the use of the silkworm BmIMP gene or recombinant plasmid in preparing a drug against silkworm nuclear polyhedrosis virus.

[0010] Preferably, the application includes: preparing BmIMP protein antibodies.

[0011] The present invention also provides the use of the BmIMP gene in preparing a transformant resistant to Bombyx mori nuclear polyhedrosis virus BmNPV.

[0012] Preferably, the application is to inhibit / silence the expression of the silkworm BmIMP gene.

[0013] Preferably, the transformant is a silkworm cell or a silkworm individual.

[0014] Compared with the prior art, the present invention has the following beneficial effects:

[0015] The present invention observed the effect of the BmIMP gene on BmNPV infecting BmN cells, and studied its function, and found that overexpression of the gene can promote the proliferation of BmNPV in BmN cells, and knocking down the gene can inhibit the replication of BmNPV in BmN cells and silkworm bodies. The present invention significantly reduces the infectivity of BmNPV in silkworm cells and silkworm bodies by knocking down BmIMP, which provides important theoretical and guiding significance for clarifying its response mechanism in the immune response of silkworms, and also provides a theoretical basis for the prevention and treatment of silkworm pus disease and provides new potential targets for drug development.

[0016] The present invention detects the tissue and induced expression spectrum of silkworm BmIMP by qRT-PCR technology, and the detection results show that the gene is specifically expressed in the hemolymph and midgut of silkworms, and the expression level is extremely high; in order to study the effect of the gene on viral infection, the present invention also constructs a recombinant plasmid of pIZT / V5-His-BmIMP, and the results after transfection into BmN cells show that it can promote the infection ability of the virus, so the BmIMP protein can be used for antibody synthesis. In addition, through the experiment of injecting siIMP in silkworms, the results show that the expression effect of the gene in silkworms is significant, and it has a significant effect on the expression of autophagy genes, which is of great value in the research and application of the immune system of silkworms, and can provide potential effective targets for the development of anti-septic drugs and virus control. As a model organism, the significant effect of the gene in silkworms can provide a reference for subsequent virus control research in other lepidopteran insects.

[0017] In addition, the test cycle of the present invention is short, the operation is simple, and the observation is convenient; the present invention has high sensitivity and can detect the fluorescence effect of virus-infected cells; the results obtained by verification of the method of the present invention are consistent with the experimental conditions of silkworm bodies and are highly credible. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 This is a diagram showing the expression of the BmIMP gene in the tissues of the silkworm.

[0019] Figure 2 This is a diagram showing the replication analysis of the virus amount in the midgut tissue of silkworms at different times after infection with BmNPV.

[0020] Figure 3 The fluorescence images were observed under an inverted fluorescence microscope; the control group was transfected with pIZT / V5-His empty plasmid, and the BmIMP group was transfected with pIZT / V5-His-BmIMP plasmid.

[0021] Figure 4 This is a graph showing the changes in the transcription level of the gene 48 hours after overexpression of the BmIMP gene.

[0022] Figure 5 This is a graph showing the changes in the replication analysis of the virus amount after adding BmNPV at different times after overexpressing the BmIMP gene.

[0023] Figure 6 This is a graph showing the changes in gene transcription levels after knocking down the BmIMP gene in BmN cells for 48 hours.

[0024] Figure 7 This is a graph showing the changes in gene transcription levels 48 hours after injection of the BmIMP gene into the silkworm.

[0025] Figure 8This is a diagram showing the effect of knocking down the BmIMP gene in silkworms on the expression levels of 14 key autophagy genes; the autophagy genes are Atg1, Atg2, Atg3, Atg4, Atg5, Atg6, Atg7, Atg8, Atg9, Atg11, Atg12, Atg13, Atg16, and Atg18. DETAILED DESCRIPTION

[0026] The present invention will be further described below in conjunction with the accompanying drawings and specific embodiments, but the protection scope of the present invention is not limited thereto. It will be appreciated by those skilled in the art that the following examples are only used to illustrate the present invention and should not be considered as limiting the scope of the present invention. Where specific techniques or conditions are not indicated in the embodiments, the techniques or conditions described in the literature in this area or the product specifications are followed. The reagents or instruments used are not indicated by the manufacturer and are conventional products that can be purchased on the market.

[0027] The silkworm used in the following examples is Bombyx mori p50, which is sourced from the Sericulture Research Institute of Jiangsu University of Science and Technology, Zhenjiang City, Jiangsu Province.

[0028] Example 1: Acquisition of silkworm BmIMP gene and construction of recombinant vector

[0029] Design of primers specific to the coding region of the silkworm BmIMP gene based on the silkworm genome sequence:

[0030] Forward primer F: 5'-TACATATGCACCCTACAAACAAATTGGC-3' (SEQ ID NO. 3),

[0031] Reverse primer R: 5'-TACTCGAGATTTAATTGTGTAACAGC-3' (SEQ ID NO. 4),

[0032] Bombyx mori p50 was obtained from the Sericulture Research Institute of Jiangsu University of Science and Technology in Zhenjiang, Jiangsu Province. The cDNA of different tissues such as the head, fat body, midgut, hemolymph and Malpighian tubules of the fifth-instar one-day silkworm was used as a template for PCR amplification. The PCR reaction conditions were: pre-denaturation at 94°C for 3 min, then denaturation at 94°C for 30 s, annealing at 60°C for 30 s, and extension at 72°C for 30 s, for a total of 35 cycles, and finally extension at 72°C for 7 min. The amplified PCR products were analyzed by agarose gel electrophoresis and the target fragments were recovered and purified using a gel recovery kit (OMEGA).

[0033] The recovered PCR product was connected to the pMD19-T vector (purchased from TaKaRa), transformed into DH5α competent cells (purchased from Vazyme), and the positive clones were obtained and sent to Shanghai Sangon Biotechnology Co., Ltd. for sequencing. The sequencing results showed that the CDS sequence of the BmIMP gene was successfully cloned, and a T clone containing SEQ ID NO.1 was obtained.

[0034] SEQ ID NO.1

[0035] ATGTGTTGCCCCGAGTTTATAGCTAAATATGTACTCTTCATCGCCAACCTTGTTTTTTCGGATCCACAGTCACGGCAATTTACAAGTACAGACCCAAACGATGAGGACTTGGAGCCAAGCATTCTCCTGTCTGAGTTGGCAGGTCTAGCTATTATACGTCTTGGAGTGGCCGTCTTAAGAAATCTACGGGATCTTCAAGATATTCTACCAGTCAACGCTTTGCCAATAGGGATAATCGTCCTCGGATGCATCATATTCATCATTGCATTCCTAGCGTGCTGTGGAGCGATAAAGGAATCAAGATGTATGCTGATAACCTACTCCATATTCATGGTGATACTGGTGGCCGTGAAGATCTACCTGGCTATCGTGGTATTCGGTTTTCTGTCCGACGTCACCAGCACAATAACCAGCTGGGTGACCACAGCGTTCAACACGAGCAGCTTAAGAGACGTATATCACGTCATGGAAGCTCTATTCAATTGCTGTGGAACAACCGGCCCCTCTTCGTACGACGGTATCTTGTCACAGCTGCCGCCCTCGTGCTGCGCCAGCCCCGTTGACAACACCTTCTATGCCCCCAACGCGTTCCCCGGCTGCACCACTCGACTGATTGATTACTTCGATACCTTCGGTAGGGCCATCGGCAGCGTTCTGATCATTATCATATTTCTTGAGCTCGTCGCCGCCAGATGTCACGCGCCTCTCACCGCGCCTCGCAATGCACCGCAAAACGCACCGCCAGTAGCTGTGGCATACTTCATAGGTTGTCTACCATTTGTTTTCGCCGCTCCCGCTTGCCGCGCCTCACGACGCACCGCAATTCGCACCGCTCTCGGCTGTGGCCAAGGGTTCATCTCGATCATCTTCGCTTGGTTCCTGTCGCATTCTTATTCGCAGAAACGGCGCGGGAATATAGCATAA。

[0036] The obtained T clone containing the target gene and the pIZT / V5-His empty plasmid were double-digested with KpnⅠ and XbaⅠ, respectively, and the target fragments were recovered using a gel recovery kit (OMEGA), ligated with T4 ligase and transformed into BL21 competent cells. After obtaining the positive clone, it was sent to Shanghai Sangon Biotechnology Co., Ltd. for sequencing to obtain the correct recombinant plasmid, which was recorded as pIZT / V5-His-BmIMP.

[0037] The positive clones were cultured in LB medium with kanamycin resistance at 37°C overnight, and the bacteria were inoculated at 1:50 for large-scale culture at 37°C. 600 =0.6-0.8, add IPTG to a final concentration of 0.1 mmol / L, continue culturing at 20°C for 8 h, collect the cells by centrifugation at 4°C, 4000 rpm for 10 min, purify the target protein in the cells with His tag, and perform purification according to the pET instructions to obtain a purified protein as shown in SEQ ID NO.2, i.e., BmIMP protein, which can be used for subsequent antibody preparation.

[0038] SEQ ID NO.2

[0039] MCCPEFIAKYVLFIANLVFSDPQSRQFTSTDPNDEDLEPSILLSELAGLAIIRLGVAVLRNLRDLQDILPVNALPIGIIVLGCIIFIIAFLACCGAIKESRCMLITYSIFMVILVAVKIYLAIVVFGFLSDVTSTITSWVTTAFNTSSLRDVYH VMEALFNCCGTTGPSSYDGILSQLPPSCCASPVDNTFYAPNAFPGCTTRLIDYFDTFGRAIGSVLIIIIFLELVAARCHAPLTAPRNAPQNAPPVAVAYFIGCLPFVFAAPACRASRRTAIRTALGCGQGFISIIFAWFLSSHSYSQKRRGNIA.

[0040] Example 2: Detection of the induced expression profile of the BmIMP gene

[0041] (1) Tissue expression of BmIMP gene:

[0042] RNA from the midgut, fat body, hemolymph, head, Malpighian tubules and other tissues of 5-year-old 1-day silkworms was extracted, and the cDNA after reverse transcription was used as a template. qRT-PCR detection was performed with gene-specific primers BmIMP-qF: GAGGACTTGGAGCCAAGC, BmIMP-qR: GATTCCTTTATCGCTCCACAGC. The conditions for qRT-PCR amplification were: 95℃ pre-denaturation for 15min, 95℃ denaturation for 15s, 60℃ annealing, extension for 30s, and 40 cycles. The obtained Ct value can be used to calculate the copy number of DNA, with GAPDH gene as the internal reference and 2 -ΔΔt To calculate the expression difference of the target gene in the sample to be tested, so as to detect the expression of BmIMP in various silkworm tissues. The test results are as follows Figure 1 As shown in the figure, it can be seen that the expression level of BmIMP gene in the hemolymph and midgut of silkworm is significantly higher than that in other tissues, so the midgut was selected for further experiments.

[0043] (2) Induction expression of BmIMP gene:

[0044] The silkworms on the first day of the fifth instar were fasted for 24 h and fed with 5 μL BmNPV (1.0×10 5 OB / mL) / fetus were infected with BmNPV, and then the larvae were collected 24h, 48h, 72h and 96h after BmNPV infection. The midguts of all larvae were dissected and mixed together to minimize individual genetic differences, and then immediately frozen in liquid nitrogen and stored at -80°C.

[0045] Extract midgut RNA, use the reverse transcribed cDNA as template, and perform qRT-PCR detection with gene-specific primers BmIMP-qF: GAGGACTTGGAGCCAAGC (SEQ ID NO.5), BmIMP-qR: GATTCCTTTATCGCTCCACAGC (SEQ ID NO.6). The conditions for qRT-PCR amplification are: 95℃ pre-denaturation for 15min, 95℃ denaturation for 15s, 60℃ annealing, extension for 30s, 40 cycles. Using GAPDH gene as internal reference, the transcription level of BmIMP gene was detected. The detection results are as follows: Figure 2 As shown in the figure, it can be seen that in the midgut tissues infected with BmNPV at different times, the virus expression level increased significantly 24 hours after virus infection, while the virus expression level was inhibited 48 hours, 72 hours and 96 hours after infection.

[0046] In summary, the relative expression level of BmIMP in the silkworm's immune tissues such as hemolymph and midgut was high, and its transcription level increased significantly 24 hours after BmNPV infection, indicating that it may play a role in the silkworm's immune response against BmNPV.

[0047] Example 3: Detection of inducible expression profile of overexpression and in vitro knockdown of BmIMP gene

[0048] (1) Gene overexpression at the cellular level:

[0049] In order to determine the expression effect of BmIMP in BmN cells (ovarian cell line cultured from the Sericulture Research Institute of Jiangsu University of Science and Technology in Zhenjiang, Jiangsu Province), the empty vector and BmIMP were transfected into BmN cells, and the cells were collected and RNA was extracted after 48 hours. The cDNA after reverse transcription was used as a template, and the gene-specific primers BmIMP-qF: GAGGACTTGGAGCCAAGC, BmIMP-qR: GATTCCTTTATCGCTCCACAGC were used for qRT-PCR detection. The conditions for qRT-PCR amplification were: 95℃ pre-denaturation for 15min, 95℃ denaturation for 15s, 60℃ annealing, extension for 30s, and 40 cycles. GAPDH was used as an internal reference to detect the transcription level of BmIMP. The detection results are shown in Figure 2. Figure 4 As shown in the figure, it can be seen that the transcription level of BmIMP is significantly increased compared with the control group, which indicates that BmIMP can be efficiently expressed in BmN cells.

[0050] In order to study the ability of BmIMP to infect BmNPV, the pIZT / V5-His-BmIMP vector was transfected into BmN cells with BmN cells transfected with empty plasmid as positive control. After 24 h, 4 μL culture medium (1×10 8 pfu / mL), and the replication of BmNPV was recorded under a fluorescence microscope at 24h, 48h, and 72h after inoculation of BV-EGFP. Figure 3 As shown in the figure, it can be seen that at 48h and 72h after virus addition, the fluorescence intensity of the experimental group was higher than that of the control group, which indicates that overexpression of BmIMP can promote the infection of the virus to cells.

[0051] In order to further explore the effect of BmIMP on BmNPV infection, the BmNPV nucleocapsid protein VP39 gene was selected in this step. The cDNA reverse transcribed 48 hours after transfection was used as a template, and GAPDH was used as an internal reference. The qRT-PCR method was used to detect the infection of the virus at different times. The conditions for qRT-PCR amplification were: 95°C pre-denaturation for 15 min, 95°C denaturation for 15 s, 60°C annealing, extension for 30 s, and 40 cycles. The detection results are shown in Figure 2. Figure 5 shown.

[0052] from Figure 5It can be seen that the virus was added 24 hours after BmIMP transfection. At 24 hours, 48 ​​hours and 72 hours after addition, the virus expression showed an upward trend compared with the control group without virus addition, among which the result at 48 hours was the most significant, further demonstrating that BmIMP can enhance the virus's ability to infect BmN cells, and the virus's ability to infect BmN cells can be inhibited by inhibiting the expression of BmIMP.

[0053] (2) Gene knockdown experiments at the cellular and individual levels:

[0054] In order to study the role of BmIMP in BmNPV infection, the disordered single-stranded small RNA (siNC) and the small interfering RNA (siRNA) of BmIMP designed by Genema were transfected into BmN cells and poisoned 24 hours later. The cells were collected and RNA was extracted 48 hours after transfection. The reverse transcribed cDNA was used as a template and the gene-specific primers BmIMP-qF: GAGGACTTGGAGCCAAGC and BmIMP-qR: GATTCCTTTATCGCTCCACAGC were used for qRT-PCR detection. The amplification conditions were: 95℃ pre-denaturation for 15min, 95℃ denaturation for 15s, 60℃ annealing, extension for 30s, and 40 cycles. GAPDH was used as an internal reference to detect the transcription level of siIMP. The detection results are shown in the figure. Figure 6 shown.

[0055] from Figure 6 It can be seen that compared with the control group, the transcription level of the siIMP experimental group at 48h was significantly reduced, which indicates that under the induction of the virus, knocking down the gene has a significant effect on reducing expression in cells.

[0056] In order to further verify the effect of BmIMP on silkworms infected by BmNPV, this step also conducted a gene knockdown experiment at the individual level. The specific steps are as follows:

[0057] Select p50 five-day-old silkworms. In order to reduce the impact of external conditions on the experiment, silkworm bodies of the same size should be selected. NC and siIMP were injected into the silkworm bodies respectively. After 24 hours of injection, the silkworms were fed with mulberry leaves added with BmNPV liquid. The silkworm bodies were collected 48 hours after feeding the virus, and they were ground and RNA was extracted. The reverse transcribed cDNA was used as a template, and the gene-specific primers BmIMP-qF: GAGGACTTGGAGCCAAGC and BmIMP-qR: GATTCCTTTATCGCTCCACAGC were used for qRT-PCR detection. The amplification conditions were: 95℃ pre-denaturation for 15min, 95℃ denaturation for 15s, 60℃ annealing, extension for 30s, and 40 cycles. GAPDH was used as an internal reference to detect changes in the transcription level of BmIMP. The results are as follows: Figure 7As shown in the figure, compared with the control group, the transcription level of the siIMP treatment group at 48h was significantly reduced, which indicates that under the induction of the virus, knocking down the gene has a significant effect of reducing expression in the silkworm body.

[0058] In summary, under viral induction, knocking down BmIMP can significantly inhibit its expression level.

[0059] (3) Detection of BmIMP gene induced expression profile:

[0060] In order to study the pathway of BmIMP in the process of BmNPV infection of silkworms, the silkworm cDNA collected in step (2) was used as a template, and the specific primers of the silkworm key autophagy genes Atg1, Atg2, Atg3, Atg4, Atg5, Atg6, Atg7, Atg8, Atg9, Atg11, Atg12, Atg13, Atg16, and Atg18 were used for qRT-PCR detection. The amplification conditions were: 95℃ pre-denaturation for 15min, 95℃ denaturation for 15s, 60℃ annealing, extension for 30s, and 40 cycles. GAPDH was used as an internal reference to detect the transcription level of autophagy-related genes. Figure 8 shown.

[0061] Among them, the specific primers of the key autophagy genes of silkworm are shown in Table 1.

[0062] Table 1. Specific primers for key autophagy genes in silkworm

[0063] BmAtg1-qF CATCGTCCACCGTGACTTGA(SEQ ID NO.7) BmAtg1-qR GTCTGCTTTGGCGTCGTATTT(SEQ ID NO.8) BmAtg2-qF GACGACTCACCGATTTACTTCAGA(SEQ ID NO.9) BmAtg2-qR CTCAGTGCCCAACAATCCAAG(SEQ ID NO.10) BmAtg3-qF AACTCAAAGCCGATAAGAAACA(SEQ ID NO.11) BmAtg3-qR TTTTAGCGTGATCTTGGGAC(SEQ ID NO.12) BmAtg4-qF TACCTCAGGGTGTATCATCA(SEQ ID NO.13) BmAtg4-qR TAAGTCTGTATCGCTGTCTTG(SEQ ID NO.14) BmAtg5-qF AAGTTCCCTGAAGACATTCT(SEQ ID NO.15) BmAtg5-qR ATTTTGTAATCCAAGCCATA(SEQ ID NO.16) BmAtg6-qF GGGCTTTTGTCTTCCGTA(SEQ ID NO.17) BmAtg6-qR TGTGGCTCAGATTTGTCCT(SEQ ID NO.18) BmAtg7-qF GAGGCGAGATGGCTGC(SEQ ID NO.19) BmAtg7-qR CGAGGTGCTAATTCCGTG(SEQ ID NO.20) BmAtg8-qF AAGGCTAGGCTTGGAGAC(SEQ ID NO.21) BmAtg8-qR CAGATGTGGGTGGAATGA(SEQ ID NO.22) BmAtg9-qF TTTAGGTGGATTGGCTGA(SEQ ID NO.23) BmAtg9-qR TTGGCACTTCGTGGATG(SEQ ID NO.24) BmAtg11-qF TAAGTCCCATAGTAGAGC(SEQ ID NO.25) BmAtg11-qR ACAAACTTCCACTTCAT(SEQ ID NO.26) BmAtg12-qF AGACGCAGAGCCAATCA(SEQ ID NO.27) BmAtg12-qR AACTCCATAATCCATCCAATA(SEQ ID NO.28) BmAtg13-qF AGAGTTTACAGTGGCGAG(SEQ ID NO.29) BmAtg13-qR TGTCTGAACGGTTAGGAG(SEQ ID NO.30) BmAtg16-qF CACATTCGGTAGAGTTAGTTTCGG(SEQ ID NO.31) BmAtg16-qR CGGCATACACTTCTCCATCGT(SEQ ID NO.32) BmAtg18-qF TTGCTGTTGGCGGTCT(SEQ ID NO.33) BmAtg18-qR CAAGCGATATGGCGGA(SEQ ID NO.34)

[0064] from Figure 8 It can be seen that 48 hours after the silkworm was injected with siIMP, the expression level of autophagy genes in the silkworm was significantly reduced, which indicates that the autophagy genes all have effects in the silkworm, and BmIMP may affect the proliferation of BmNPV through the autophagy pathway.

[0065] In summary, BmIMP can be efficiently expressed in the immune tissues of silkworms and may play a role in the immune response against BmNPV. Overexpression of BmIMP can promote the infectivity of BmNPV, while knockdown inhibits the infectivity of BmNPV. BmIMP may regulate the proliferation of BmNPV in silkworms by promoting the expression of autophagy genes.

[0066] The embodiments are preferred implementations of the present invention, but the present invention is not limited to the above-mentioned implementations. Any obvious improvements, substitutions or modifications that can be made by those skilled in the art without departing from the essential content of the present invention belong to the protection scope of the present invention.

Claims

1. A silkworm BmIMP gene capable of regulating BmNPV infection of BmN cells, It is characterized in that The nucleotide sequence of the silkworm BmIMP gene coding region is shown in SEQ ID NO.

1.

2. The silkworm BmIMP gene capable of regulating BmNPV infection of BmN cells according to claim 1, It is characterized in that The amino acid sequence of the protein encoded by the silkworm BmIMP gene is shown in SEQ ID NO.

2.

3. The silkworm BmIMP gene capable of regulating BmNPV infection of BmN cells according to claim 1, It is characterized in that Overexpression of the silkworm BmIMP gene can promote the proliferation of BmNPV in BmN cells, and inhibition / silencing of the expression of the silkworm BmIMP gene can inhibit the infection ability of BmNPV in silkworm cells and silkworm bodies.

4. A recombinant plasmid, It is characterized in that The recombinant plasmid comprises the coding region nucleotide sequence of the silkworm BmIMP gene according to claim 1.

5. Use of the silkworm BmIMP gene according to claim 1 or the recombinant plasmid according to claim 4 in the preparation of an anti-Bombyx mori nuclear polyhedrosis virus drug.

6. The use according to claim 5, It is characterized in that The application includes: preparing BmIMP protein antibodies.

7. Use of the BmIMP gene according to claim 1 in preparing a transformant resistant to Bombyx mori nuclear polyhedrosis virus (BmNPV).

8. The use according to claim 7, It is characterized in that The application is to inhibit / silence the expression of silkworm BmIMP gene.

9. The use according to claim 7, It is characterized in that The transformant is a silkworm cell or a silkworm individual.

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