CN116250548A - Application of throat-inhibiting side body neuropeptides in preventing and treating periplaneta americana - Google Patents

Abstract

The invention discloses application of a throat-inhibiting lateral neuropeptide in preventing and treating periplaneta americana, wherein AST comprises AST1 (SEQ ID NO. 1) and AST2 (SEQ ID NO. 2). According to the invention, the female adult periplaneta americana in the first reproductive cycle is injected with AST1 and AST2, so that the synthesis of vitellogenin Vg can be inhibited, JH synthesis in pharyngeal side bodies is inhibited, thereby inhibiting JH signals of fat bodies, further inhibiting vitellization of the fat bodies, and finally inhibiting ovarian maturity; the purpose of preventing and controlling the American cockroach can be achieved by controlling the reproduction of females. The method has high specificity, can directly target the reproduction of female American cockroaches, does not generate harmful substances polluting the environment, and provides a new method for pest control.

Description

Application of throat-inhibiting side body neuropeptides in preventing and treating periplaneta americana

Technical Field

The invention belongs to the technical field of biological control, and particularly relates to application of throat-inhibiting lateral nerve peptide in control of periplaneta americana.

Background

Female insect reproduction has received widespread attention due to its importance in species reproduction and its great potential as a target for pest control. Insects are the most abundant and diverse species in the world, and the presently known species have exceeded 100 tens of thousands. Insects are of a great variety and are extremely strong in adaptability, and the figures of activities of the insects can be seen from the tropical region with high temperature to the cold polar region. The insects are so widely distributed that, in addition to having a strong adaptability, they have a strong reproduction ability. In most insects, including incompletely metamorphosed insects, reproduction is dominated by JH, which acts as gonadotropin to regulate the maturation of these insect reproductive organs (Roy et al, 2018). In the adult stage of insects, JH activates the expression of its primary response gene Kr-h1 by binding to the receptor Met, induces synthesis of Vg in fat bodies, promotes absorption of Vg by ovaries and development of follicular cells, and thereby exerts a function of promoting reproduction (Roy et al, 2018; zhu et al, 2020).

The American cockroach (Periplaneta americana) belongs to the order of Blatta, and insects of the family Blattaceae, commonly called cockroaches, have very strong environment adaptation capability and fertility, and are recognized indoor sanitary pests in the world. It can transmit pathogenic factors such as bacteria, protozoa, viruses, etc.; as an allergen, causes allergic reactions in humans. With global warming, the urban process is accelerated, traffic and trade are rapidly developed, the hazard degree is more and more serious, has become important sanitary pests for hotels, restaurants, families, hospitals, schools, food processing, selling, catering and other units. The periplaneta americana belongs to incomplete metamorphosis insects, has strong fertility and environmental adaptability, can perform parthenogenesis, transmits pathogenic microorganisms, and is a serious urban sanitary pest. At present, chemical control is still a main control mode for controlling the population quantity of periplaneta americana, and long-term abuse of organophosphorus chemical pesticides, carbamate chemical pesticides and pyrethroid chemical pesticides tends to lead to improvement of pest resistance and environmental pollution. Therefore, it is imperative to establish a safe and effective cockroach control mechanism.

Disclosure of Invention

The invention aims to provide application of AST1 and AST2 proteins in preventing and treating periplaneta americana.

The technical scheme adopted by the invention is as follows:

in a first aspect of the invention there is provided the use of a protein in at least one of A1) to A14) as follows; the amino acid sequence of the protein is shown as SEQ ID NO.1 and/or SEQ ID NO. 2;

a1 Inhibiting expression of insect vitellogenin genes Vg1 and/or Vg 2;

a2 Preparing a product for inhibiting the expression of the insect vitellogenin genes Vg1 and/or Vg 2;

a3 Inhibiting insect ovarian maturation;

a4 Preparing a product for inhibiting the maturation of the ovaries of the insects;

a5 Inhibiting insect ovarian development;

a6 Preparing a product for inhibiting the development of insect ovaries;

a7 Inhibiting insect ovarian follicular cell development;

a8 Preparing a product for inhibiting development of ovarian follicular cells of the insect;

a9 Inhibiting expression of juvenile hormone;

a8 Preparing a product that inhibits juvenile hormone expression;

a9 Inhibiting female reproduction of insects;

a10 Preparing a product for inhibiting reproduction of female insects;

a11 A) controlling pests;

a12 Preparing a product for controlling pests;

a13 Reducing the reproductive capacity of insects;

a14 A product that reduces the reproductive capacity of insects is prepared.

In a second aspect of the invention there is provided the use of a substance to increase the protein content and/or activity of the first aspect of the invention in at least one of A1) to A14);

a1 Inhibiting expression of insect fat body vitellogenin genes Vg1 and/or Vg 2;

a2 Preparing a product for inhibiting the expression of insect fat body vitellogenin genes Vg1 and/or Vg 2;

a3 Inhibiting insect ovarian maturation;

a4 Preparing a product for inhibiting the maturation of the ovaries of the insects;

a5 Inhibiting insect ovarian development;

a6 Preparing a product for inhibiting the development of insect ovaries;

a7 Inhibiting insect ovarian follicular cell development;

a8 Preparing a product for inhibiting development of ovarian follicular cells of the insect;

a9 Inhibiting expression of juvenile hormone;

a8 Preparing a product that inhibits juvenile hormone expression;

a9 Inhibiting female reproduction of insects;

a10 Preparing a product for inhibiting reproduction of female insects;

a11 A) controlling pests;

a12 Preparing a product for controlling pests;

a13 Reducing the reproductive capacity of insects;

a14 A product that reduces the reproductive capacity of insects is prepared.

In a third aspect of the present invention there is provided the use of a substance which increases expression of a gene encoding a protein as described in the first aspect of the present invention in at least one of A1) to A14);

a1 Inhibiting expression of insect fat body vitellogenin genes Vg1 and/or Vg 2;

a2 Preparing a product for inhibiting the expression of insect fat body vitellogenin genes Vg1 and/or Vg 2;

a3 Inhibiting insect ovarian maturation;

a4 Preparing a product for inhibiting the maturation of the ovaries of the insects;

a5 Inhibiting insect ovarian development;

a6 Preparing a product for inhibiting the development of insect ovaries;

a7 Inhibiting insect ovarian follicular cell development;

a8 Preparing a product for inhibiting development of ovarian follicular cells of the insect;

a9 Inhibiting expression of juvenile hormone;

a8 Preparing a product that inhibits juvenile hormone expression;

a9 Inhibiting female reproduction of insects;

a10 Preparing a product for inhibiting reproduction of female insects;

a11 A) controlling pests;

a12 Preparing a product for controlling pests;

a13 Reducing the reproductive capacity of insects;

a14 A product that reduces the reproductive capacity of insects is prepared.

In some embodiments of the invention, the substance that increases the expression of the gene, the substance that increases the activity of the protein or the substance that increases the content of the protein is a protein according to the first aspect of the invention and/or a biological material related to a protein according to the first aspect of the invention, which biological material is any one of the following B1) to B4):

b1 A nucleic acid molecule encoding a protein according to the first aspect of the invention;

b2 An expression cassette comprising the nucleic acid molecule of B1);

b3 A recombinant vector comprising the nucleic acid molecule of B1) or a recombinant vector comprising the expression cassette of B1);

b4 A recombinant microorganism comprising the nucleic acid molecule of B1), a recombinant microorganism comprising the expression cassette of B2), or a recombinant microorganism comprising the recombinant vector of B3).

In a fourth aspect of the invention, there is provided a method comprising increasing the content and/or activity of a protein according to the first aspect of the invention in an insect; the method is used for at least one of C1) to C8):

c1 Inhibiting expression of insect fat body vitellogenin genes Vg1 and/or Vg 2;

c2 Inhibiting insect ovarian maturation;

c3 Inhibiting insect ovarian development;

c4 Inhibiting insect ovarian follicular cell development;

c5 Inhibiting expression of juvenile hormone;

c6 Inhibiting female reproduction of insects;

c7 A) controlling pests;

c8 Reducing the reproductive capacity of insects.

In some embodiments of the invention, the method of increasing the content and/or activity of a protein according to the first aspect of the invention in an insect is by introducing a protein according to the first aspect of the invention or a protein-related biomaterial according to the first aspect of the invention into the insect.

In some embodiments of the invention, the biomaterial is any one of the following B1) to B4):

b1 A nucleic acid molecule encoding a protein according to the first aspect of the invention;

b2 An expression cassette comprising the nucleic acid molecule of B1);

b3 A recombinant vector comprising the nucleic acid molecule of B1) or a recombinant vector comprising the expression cassette of B1);

b4 A recombinant microorganism comprising the nucleic acid molecule of B1), a recombinant microorganism comprising the expression cassette of B2), or a recombinant microorganism comprising the recombinant vector of B3).

In some embodiments of the invention, the means of introduction comprises injection.

In some embodiments of the invention, the insect is a female adult in the first reproductive cycle.

In a fifth aspect, the invention provides the use of a method according to the fourth aspect of the invention for controlling pests.

In some embodiments of the invention, the pest or insect is american cockroach.

In a sixth aspect of the invention, there is provided a composition comprising D1) and D2);

d1 Protein with a sequence shown as SEQ ID NO.1 or protein related biological material shown as SEQ ID NO. 1;

d2 A protein with a sequence shown as SEQ ID NO.1 or a protein related biological material shown as SEQ ID NO. 1.

In some embodiments of the invention, the relevant biomaterial is any one of the following B1) to B4):

b1 A nucleic acid molecule encoding said protein;

b2 An expression cassette comprising the nucleic acid molecule of B1);

b3 A recombinant vector comprising the nucleic acid molecule of B1) or a recombinant vector comprising the expression cassette of B1);

b4 A recombinant microorganism comprising the nucleic acid molecule of B1), a recombinant microorganism comprising the expression cassette of B2), or a recombinant microorganism comprising the recombinant vector of B3).

The beneficial effects of the invention are as follows:

according to the invention, the AST polypeptides AST1 and AST2 are injected into the female periplaneta americana adults in the first reproductive cycle, so that the synthesis of vitellogenin Vg can be inhibited, and JH synthesis in pharyngeal side bodies is inhibited to inhibit JH signals of fat bodies, further inhibit vitelline generation of the fat bodies, and finally inhibit ovarian maturation; the purpose of preventing and controlling the American cockroach can be achieved by controlling the reproduction of females. The method has high specificity, can directly target the reproduction of female American cockroaches, does not generate harmful substances polluting the environment, and provides a new method for pest control.

Drawings

FIG. 1 shows the results of detecting the expression level of the fat body vitellogenin genes Vg1 and Vg2 after AST injection; wherein fig. 1A is injection AST1 and fig. 1B is injection AST2.

Fig. 2 shows changes in ovarian morphology following AST injection; wherein fig. 2A is the change in ovarian phenotype following AST injection and fig. 2B is the change in ovarian first grain egg length following AST injection.

Figure 3 is the change in Vg in ovaries after AST injection; fig. 3A shows the change of total protein content of ovaries after AST injection, fig. 3B shows the characteristic sequence of Vg1 identified by mass spectrometry, and fig. 3C shows the characteristic sequence of Vg2 identified by mass spectrometry.

FIG. 4 shows morphological changes of follicular cells and patent after AST injection.

FIG. 5 shows JH synthesis and changes in JH signal after AST injection; wherein fig. 5A is injection AST1 and fig. 5B is injection AST2.

Wherein P <0.001; * P <0.01; * P <0.05.

Detailed Description

The conception and the technical effects produced by the present invention will be clearly and completely described in conjunction with the embodiments below to fully understand the objects, features and effects of the present invention. It is apparent that the described embodiments are only some embodiments of the present invention, but not all embodiments, and that other embodiments obtained by those skilled in the art without inventive effort are within the scope of the present invention based on the embodiments of the present invention.

Example 1 AST design and Synthesis

The precursor protein of the pharyngeal side body neuropeptide (AST) needs to be cut into short mature polypeptides by enzyme digestion to play a corresponding function. Mature AST sequences (SEQ ID No.1 and SEQ ID No. 2) were designed based on the American cockroach AST precursor sequence (NCBI database ID: CAA 62500.1). The AST precursor protein sequence is submitted to on-line neuropeptide analysis prediction software http:// stagbete. Animal. Uiuc. Edu/cgi-bin/neurored. Py, to predict signal peptide and cleavage sites. And determining the sequence of the mature neuropeptide after cutting according to the sequence between enzyme cutting sites. Mature AST polypeptides (SEQ ID No.1 and SEQ ID No. 2) were synthesized in the C-terminal to N-terminal direction using a linear polypeptide synthesis method (Ai Ji organism, guangzhou, china).

Mature American cockroach AST polypeptide sequence 1 (SEQ ID No. 1): SPSGMQRLYGFGL-NH ₂ ；

Mature American cockroach AST polypeptide sequence 2 (SEQ ID No. 2): ADGRLYAFGL-NH ₂ 。

EXAMPLE 2 AST inhibition of female American cockroach reproduction

(1) AST injection and phenotypic observation statistics

AST1 and AST2 were diluted with sterilized ultrapure water to 50. Mu.g/. Mu.L of the working solution, respectively. Female periplaneta americana healthy on day 2 after eclosion was picked up, fully anesthetized with carbon dioxide, and then injected with AST1 and AST2, respectively, into the second body node of the periplaneta americana abdomen with a microinjector, each of which was injected with 100 μg (2 μl,50 μg/μl). The control group was injected with sterilized ultrapure water. Injections were given on days 2 and 4, respectively, 2 total injections. Wherein 30 injections were injected per treatment. On day 5, head and fat bodies are respectively taken, every 4 tissues of periplaneta americana are combined into one sample, the sample is quickly frozen in liquid nitrogen, and then the sample is preserved at the temperature of minus 80 ℃ for standby, and is used for total RNA extraction, and four groups of samples are taken for each treatment; combining the tissues of every 4 American cockroaches into one sample, putting the samples into liquid nitrogen for quick freezing, and then preserving the samples at the temperature of-80 ℃ for standby, wherein the samples are used for total protein extraction, and four groups of samples are taken for each treatment; the ovaries were taken for photographing, and after DAPI and Phalloidin staining, confocal microscopy was used for photographing, and four pairs of ovaries were photographed, respectively. Experiments were repeated 3 times.

(2) Effect of AST on expression of the fat body vitellogenin genes Vg1 and Vg2

According to the gene sequence in the American cockroach genome database (Li et al, 2018), a Primer design software Primer5 is used for designing fluorescent quantitative PCR Primer, and the length of amplified fragment is about 80-150bp. Primers used for fluorescent quantitative PCR detection:

Vg1-FP：TGCTGATGAGGACACAACCT(SEQ ID No.3)；

Vg1-RP：CCACTTGCTTCACTGGATCG(SEQ ID No.4)；

Vg2-FP：GAGAAGCAGACACGAGGAACG(SEQ ID No.5)；

Vg2-RP：CCCTTGAACGCCACTGAGAC(SEQ ID No.6)；

actin-FP：CATCCTGCGTTTGGATCTGG(SEQ ID No.7)；

actin-RP：TTTCTCGTTCGGCAGTGGTG(SEQ ID No.8)。

using HieffTM qPCR

Green Master Mix (Low Rox Plus) reagent, operated according to its instructions, was subjected to fluorescent quantitative PCR detection. Comparing the two groups of data, calculating the relative expression quantity of the target gene by using housekeeping gene actin, and then using t-testAnd comparing whether a significant difference exists between the two groups of data to obtain the relative expression quantity of the target gene at a certain treatment or a certain time point. Each sample was repeated three times, each treatment was performed three times in biological replicates, the average and standard error of each set of data was taken and a chart was drawn.

After AST1 injection, detecting the expression level in fat body, and finding that the expression levels of Vg1 and Vg2 are obviously reduced by more than 80 percent (figure 1A); the expression levels of Vg1 and Vg2 were also significantly down-regulated by nearly 80% after AST2 injection (fig. 1B). These results indicate that AST inhibits expression of the fat body vitellogenin genes Vg1 and Vg 2.

(3) Observation of ovarian phenotype

After AST1 and AST2 are injected, the American cockroach is taken out, after anesthesia, the American cockroach is fixed on a wax disk by an insect needle, the wax disk is placed under an Olympus SZ61 dissecting microscope, the abdomen is cut off, the ovary is peeled off from the tissue, the American cockroach is taken out and placed in a culture dish, and the American cockroach physiological saline is added to enable the ovary to float in the solution. And continuing dissecting the ovary under a microscope, taking out the peripheral muscle tissue of the ovary, placing the ovary under a Nikon DS-Ri2 camera for photographing, and recording the development condition of the ovary. Under a microscope, ovaries were dissected into individual tubules with dissected forceps, morphology of the tubules was observed, recorded by photograph with a nikon DS-Ri2 camera, and the length of the ovaries' head grain eggs was measured using NIS-Elements BR 4.50.00 software.

Ovarian development was inhibited after AST1 and AST2 injections (fig. 2A), with a significant 63% and 61.4% reduction in head grain egg length, respectively, compared to the control (fig. 2B). The results show that the ovary maturation is inhibited after AST injection, which shows that AST has obvious inhibition effect on ovum.

(4) AST Effect on Total ovarian protein

After treatment, the American cockroach ovaries were ground in the Biyundian RIPA lysate (PMSF was added in advance at a final concentration of 1 mM), centrifuged at 12000g for 30min at 4℃and the supernatant was filtered with a 0.22 μm filter membrane. Protein concentration was measured using assist in the holy organism BCA kit and adjusted to 2ug/ul. Adding a Loading buffer into a protein sample, and carrying out boiling water bath for 5 minutes; loading 10ul of sample into each glue hole; during electrophoresis, the voltage of 80V is firstly used for 30min, and after a sample enters the separation gel, the voltage is adjusted to 130V to continue electrophoresis for 90min; taking down the gel, placing the gel in a dyeing vessel added with coomassie brilliant blue R250 dyeing liquid, placing the gel on a shaking table, setting the rotating speed to 45R/min, pouring out the dyeing liquid after dyeing for 30min, and flushing with clear water for 3 times; adding a decoloring liquid, placing on a shaking table, and pouring the decoloring liquid after a strip is clearly visible; is placed in a scanner for photographing.

The SDS-PAGE gel electrophoresis results showed a significant decrease in total ovarian protein content following AST injection (FIG. 3A). The 100kD protein band is obviously detected and analyzed by gel cutting mass spectrometry, and the band is found to be Vg, including Vg1 and Vg2, and the polypeptide sequences are EPGNLNLAR (SEQ ID No. 9) (figure 3B) and HELIAVAYALPSK (SEQ ID No. 10) (figure 3C) respectively. The results show that AST inhibits the absorption of Vg protein by ovaries, so that the accumulation of Vg of ovaries is inhibited, and the development of ovaries is inhibited.

(5) Effect of AST on ovarian follicular cell development

Taking out the American cockroach on the 5 th day after treatment, dissecting out the ovary after anesthesia, putting the ovary into a 0.5mL centrifuge tube, adding 400ul of fixing solution, and fixing the American cockroach on a shaking table at 25 ℃ for 1 hour; sucking out the fixing solution, adding 400 mu L of PBT for cleaning, pouring out the waste liquid, and repeating the cleaning for 4 times; 400ul of PBT is added, and the mixture is placed in a shaking table for fine washing for 1 hour at 25 ℃; sucking out PBT, adding 400 mu L of PBT again, adding 1ul of DAPI/phalloidin (1:10000), wrapping a centrifuge tube with tinfoil, and dyeing for 30min on a shaker in a dark place; then the mixture is washed by PBT for 4 times, 400 mu L each time; adding 400 mu L of PBT, and placing in a shaking table at 25 ℃ for fine washing for 1 hour; performing secondary dissection, taking out the tissue, placing the tissue on a glass slide, sucking the liquid, dripping a certain amount of 50% glycerol, dissecting the required tissue, sealing the tissue, observing the tissue under a confocal microscope, and photographing. Ovarian follicular cell photographs taken at 5 x magnification under a confocal microscope 40 x objective field of view were randomly selected. Each treatment randomly selects 3 photo measurements.

AST1 and AST2 treated groups showed no interstitial cell size (white arrow) and significantly decreased cells compared to the control group, which inhibited development of ovarian follicular cells after AST injection (fig. 4). The results demonstrate that AST inhibits follicular cell development and the formation of patent.

(6) Detection of JH key enzyme gene expression in pharyngeal flank and JH signal transduction gene expression in fat body

After AST1 and AST2 treatment, expression of jumt and Cyp15A1, which are essential genes for juh synthase, was significantly reduced in pharyngeal flank (fig. 5A), and Met and Kr-h1, which are essential genes for juh signal transduction, were also significantly reduced in fat body (fig. 5B). These results indicate that AST inhibits JH signaling in the fat body by inhibiting JH synthesis in the pharyngeal side body.

By combining the experimental results of fig. 1-5, the experiment shows that AST inhibits JH signals of fat bodies by inhibiting JH synthesis in pharyngeal side bodies, further inhibits vitelline formation of fat bodies, and finally inhibits ovarian maturation. The experiments prove that AST has the effect of inhibiting the reproduction of periplaneta americana females. The control of female reproduction can be realized to prevent and control American cockroach.

The present invention has been described in detail in the above embodiments, but the present invention is not limited to the above examples, and various changes can be made within the knowledge of those skilled in the art without departing from the spirit of the present invention. Furthermore, embodiments of the invention and features of the embodiments may be combined with each other without conflict.

Claims (10)

1. The use of a protein in at least one of the following A1) to A14); the amino acid sequence of the protein is shown as SEQ ID NO.1 and/or SEQ ID NO. 2;

a1 Inhibiting expression of insect vitellogenin genes Vg1 and/or Vg 2;

a2 Preparing a product for inhibiting the expression of the insect vitellogenin genes Vg1 and/or Vg 2;

a3 Inhibiting insect ovarian maturation;

a4 Preparing a product for inhibiting the maturation of the ovaries of the insects;

a5 Inhibiting insect ovarian development;

a6 Preparing a product for inhibiting the development of insect ovaries;

a7 Inhibiting insect ovarian follicular cell development;

a8 Preparing a product for inhibiting development of ovarian follicular cells of the insect;

a9 Inhibiting expression of juvenile hormone;

a8 Preparing a product that inhibits juvenile hormone expression;

a9 Inhibiting female reproduction of insects;

a10 Preparing a product for inhibiting reproduction of female insects;

a11 A) controlling pests;

a12 Preparing a product for controlling pests;

a13 Reducing the reproductive capacity of insects;

a14 A product that reduces the reproductive capacity of insects is prepared.

2. Use of a substance which increases the protein content and/or activity of claim 1 in at least one of A1) to a 14);

a1 Inhibiting expression of insect fat body vitellogenin genes Vg1 and/or Vg 2;

a2 Preparing a product for inhibiting the expression of insect fat body vitellogenin genes Vg1 and/or Vg 2;

a3 Inhibiting insect ovarian maturation;

a4 Preparing a product for inhibiting the maturation of the ovaries of the insects;

a5 Inhibiting insect ovarian development;

a6 Preparing a product for inhibiting the development of insect ovaries;

a7 Inhibiting insect ovarian follicular cell development;

a8 Preparing a product for inhibiting development of ovarian follicular cells of the insect;

a9 Inhibiting expression of juvenile hormone;

a8 Preparing a product that inhibits juvenile hormone expression;

a9 Inhibiting female reproduction of insects;

a10 Preparing a product for inhibiting reproduction of female insects;

a11 A) controlling pests;

a12 Preparing a product for controlling pests;

a13 Reducing the reproductive capacity of insects;

a14 A product that reduces the reproductive capacity of insects is prepared.

3. Use of a substance which increases the expression of a gene encoding a protein according to claim 1 in at least one of A1) to a 14);

a1 Inhibiting expression of insect fat body vitellogenin genes Vg1 and/or Vg 2;

a2 Preparing a product for inhibiting the expression of insect fat body vitellogenin genes Vg1 and/or Vg 2;

a3 Inhibiting insect ovarian maturation;

a4 Preparing a product for inhibiting the maturation of the ovaries of the insects;

a5 Inhibiting insect ovarian development;

a6 Preparing a product for inhibiting the development of insect ovaries;

a7 Inhibiting insect ovarian follicular cell development;

a8 Preparing a product for inhibiting development of ovarian follicular cells of the insect;

a9 Inhibiting expression of juvenile hormone;

a8 Preparing a product that inhibits juvenile hormone expression;

a9 Inhibiting female reproduction of insects;

a10 Preparing a product for inhibiting reproduction of female insects;

a11 A) controlling pests;

a12 Preparing a product for controlling pests;

a13 Reducing the reproductive capacity of insects;

a14 A product that reduces the reproductive capacity of insects is prepared.

4. Use according to claim 2 or 3, wherein the substance that increases the expression of the gene, the substance that increases the activity of the protein or the substance that increases the content of the protein is a protein as defined in claim 1 and/or a biological material related to a protein as defined in claim 1, which biological material is any one of the following B1) to B4):

b1 A nucleic acid molecule encoding the protein of claim 1;

b2 An expression cassette comprising the nucleic acid molecule of B1);

b3 A recombinant vector comprising the nucleic acid molecule of B1) or a recombinant vector comprising the expression cassette of B1);

b4 A recombinant microorganism comprising the nucleic acid molecule of B1), a recombinant microorganism comprising the expression cassette of B2), or a recombinant microorganism comprising the recombinant vector of B3).

5. A method comprising increasing the content and/or activity of a protein as defined in claim 1 in an insect, said method being used for at least one of C1) to C8):

c1 Inhibiting expression of insect fat body vitellogenin genes Vg1 and/or Vg 2;

c2 Inhibiting insect ovarian maturation;

c3 Inhibiting insect ovarian development;

c4 Inhibiting insect ovarian follicular cell development;

c5 Inhibiting expression of juvenile hormone;

c6 Inhibiting female reproduction of insects;

c7 A) controlling pests;

c8 Reducing the reproductive capacity of insects.

6. The method according to claim 5, wherein the method of increasing the content and/or activity of the protein according to claim 1 in an insect is to introduce the protein according to claim 1 or the protein-related biological material according to claim 1 into the insect.

7. The method according to claim 6, wherein the biological material is any one of the following B1) to B4):

b1 A nucleic acid molecule encoding the protein of claim 1;

b2 An expression cassette comprising the nucleic acid molecule of B1);

b3 A recombinant vector comprising the nucleic acid molecule of B1) or a recombinant vector comprising the expression cassette of B1);

b4 A recombinant microorganism comprising the nucleic acid molecule of B1), a recombinant microorganism comprising the expression cassette of B2), or a recombinant microorganism comprising the recombinant vector of B3).

8. The method of claim 6, wherein the introducing means comprises injection.

9. A composition comprising D1) and D2);

d1 Protein with a sequence shown as SEQ ID NO.1 or protein related biological material shown as SEQ ID NO. 1;

d2 A protein with a sequence shown as SEQ ID NO.1 or a protein related biological material shown as SEQ ID NO. 1.

10. The use according to any one of claims 1 to 4 or the method according to any one of claims 5 to 8, wherein the pest or insect is american cockroach.

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