Trichogramma japonicum juvenile hormone epoxy hydrolase recombinant protein, and preparation and application thereof
By obtaining and expressing the recombinant protein of the TjJHEH gene of the epoxy hydrolase TjJHEH gene, the problem of short life span of the epoxy hydrolase of the epoxy hydrolase of the epoxy hydrolase of the epoxy hydrolase of the epoxy hydrolase of the epoxy hydrolase of the epoxy hydrolase of the epoxy hydrolase of the epoxy hydrolase of the epoxy hydrolase of the epoxy hydrolase was solved, the life span was extended and the growth and development regulation was achieved, and its application potential in rice pest control was improved.
Patent Information
- Application Number
- CN202411826935.5
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-12-12
- Publication Date
- 2025-03-28
AI Technical Summary
Among the existing red-eyed bee breeding technologies, the lifespan of rice borer red-eyed bee is relatively short, and it is difficult to effectively prolong its growth and development cycle by adjusting the temperature, and there is a lack of relevant gene regulation technology.
The TjJHEH gene of epoxy hydrolase TjJHEH gene was obtained by PCR, and heterologous expression was performed using genetic engineering technology to obtain recombinant proteins, and the life span and growth regulation of epoxy hydrolase wasp was achieved through feeding.
It extends the lifespan of rice borer red-eyed wasp, regulates its growth and breeding process, improves its application time, and has important application potential in rice pest control.
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Abstract
Description
Technical Field
[0001] The invention belongs to the field of genetic engineering and molecular biology, and particularly relates to a recombinant protein of juvenile hormone epoxide hydrolase of Trichogrammatus oryzae, as well as its preparation and application. Background Art
[0002] With the continuous improvement of agricultural production levels, the irrational use of pesticides not only makes agricultural pests resistant, but also has a serious impact on ecological security and food safety. As an important natural enemy of pests, the rice stem borer trichogramma japonicum is an important alternative technology and product for chemical control of rice pests. It has been widely used at home and abroad and has achieved significant economic benefits. Its wide application is of great significance for reducing the application of pesticides, protecting the environment and biodiversity, and ensuring the safety of organic green agricultural product production. In recent years, the green control technology of using rice stem borer trichogramma japonicum to control rice stem borer and rice leaf folder has been successfully demonstrated on a large scale in Jilin, Liaoning, Hunan, Jiangxi and other places in my country.
[0003] The rice stem borer trichogrammatid is a trichogrammatid mainly used to control rice borers. Its body length is generally 0.44-0.59mm, the body is black to dark brown, the compound eyes are red, and the forewings have rows of longitudinal fine hairs. It is the dominant trichogrammatid population in rice fields. The hosts include a variety of insects from the Pyralidae, Lycoris, Noctuidae, Hesperidae and Marsh Fly. The reason why the rice stem borer trichogrammatid has become a natural enemy insect that is widely promoted and applied is closely related to its fast reproduction, good effect, labor saving and pollution-free. The adult trichogrammatid of the rice stem borer lays eggs in the eggs of pests (hosts), and the trichogrammatid eggs develop into larvae in the host eggs. The trichogrammatid larvae feed on the nutrients in the host eggs, so that the eggs of the pests cannot hatch normally, thereby achieving the purpose of pest control. The trichogrammatid larvae develop into adults in the host eggs and drill out of the host eggs, continuing to look for new host eggs to complete parasitism.
[0004] As the concept of green pest control has gained popularity, the use of trichogrammatids to control rice pests has been increasingly recognized, and the demand for trichogrammatids is increasing. However, the existing trichogrammatid breeding technology has a short lifespan of about 5 days. Currently, scientific and technological personnel mainly control the growth and development of trichogrammatids by adjusting the temperature to extend their shelf life. However, there is no relevant technology to explore the genes that regulate the growth and development of trichogrammatids and further regulate their growth and development.
[0005] Juvenile hormone is one of the most important hormones that regulates insect development, metamorphosis and reproduction. Juvenile hormone can inhibit the development of insect larvae and regulate the growth and reproduction of adults. Juvenile hormone deficiency in insect adults can cause reproductive diapause and prolong lifespan. Juvenile hormone epoxidehydrolase (JHEH) is one of the key enzymes that regulate the metabolism of insect juvenile hormone (JH). Figure 1 Exogenous juvenile hormone epoxide hydrolase can reduce the juvenile hormone content, prolong the life span of insect adults, and regulate reproductive capacity. Summary of the invention
[0006] The present invention provides a recombinant protein of juvenile hormone epoxide hydrolase of Trichogrammatus oryzae, preparation and application thereof, which has good application potential in controlling rice pests by using parasitic wasps and reducing the amount of pesticides applied. The TjJHEH gene of juvenile hormone epoxide hydrolase of Trichogrammatus oryzae obtained by PCR method is heterologously expressed by genetic engineering technology to obtain a recombinant protein.
[0007] The specific plan is:
[0008] A recombinant protein of juvenile hormone epoxide hydrolase of Trichogrammatus oryzae, wherein the amino acid sequence of the recombinant protein is shown in SEQ ID NO:1.
[0009] Optionally, the nucleotide sequence of the gene encoding the recombinant protein is shown in SEQ ID NO: 2.
[0010] The method for preparing the recombinant protein of Trichogramma oryzae juvenile hormone epoxide hydrolase of the present invention comprises:
[0011] S1: The nucleic acid fragment of juvenile hormone epoxide hydrolase of Trichogrammatidae was amplified by PCR reaction, and the PCR fragment was obtained after agarose gel electrophoresis.
[0012] S2: Use high-fidelity enzyme to prepare linear fragment of expression vector, and then perform homologous recombination ligation with the prepared target PCR fragment after agarose gel electrophoresis inspection and product recovery and purification, and then transfer into E. coli DH5α competent cells by heat shock method;
[0013] S3: All the screened positive transformants are transferred into the expression host Escherichia coli to perform a second round of screening of positive transformants;
[0014] S4: Inducing the expression of engineered bacteria to obtain bacterial cells;
[0015] S5: The bacterial cells are crushed, separated and purified to obtain the target protein, and the target protein is renatured to obtain the recombinant protein.
[0016] Optionally, the primers used in S1 are: forward 5'-ATGGCGTTACTTTTGTTGACGG-3', reverse 5'-CTACATCTCGGTTTTCTTCGAC-3'.
[0017] Optionally, the PCR fragment in S1 is synthesized according to the nucleotide sequence shown in SEQ ID NO:2.
[0018] Optionally, the expression vector in S2 is pET-28a, and the Escherichia coli in S3 is BL21 (DE3).
[0019] Optionally, in S4, the induction expression condition is: at a bacterial solution OD 600 When the absorbance is 0.6-0.8, add 0.4mM-1.0mM IPTG and induce at 25-40℃ for 4-8h.
[0020] The invention discloses an application of the Trichogrammatis juvenile hormone epoxide hydrolase recombinant protein in preparing a life-extending drug and / or a growth and development regulating drug for Trichogrammatis pyrifolia.
[0021] Optionally, the recombinant protein is used to extend the life span and / or regulate the growth and development of Trichogramma oryzae through feeding.
[0022] Optionally, the method for fluorescence quantitative detection of the expression level of the recombinant protein of juvenile hormone epoxide hydrolase of Trichogrammatis oryzae in adults of Trichogrammatis oryzae comprises the following steps:
[0023] S1: RNA from the 1st, 2nd and 3rd day Trichogrammatidae adults was extracted separately and reverse transcribed to obtain cDNA;
[0024] S2: The expression level of TjERP gene of 1-3 day old Trichogrammatidae adults was detected by fluorescence quantitative PCR reaction.
[0025] The beneficial effects obtained by the present invention are:
[0026] (1) A TjJHEH gene of juvenile hormone epoxide hydrolase of Trichogrammatus oryzae was obtained by PCR. Its amino acid sequence is shown in SEQ ID NO.1, and its nucleotide sequence is shown in SEQ ID NO:2.
[0027] (2) The present invention expresses the TjJHEH recombinant protein of Trichogramma oryzae juvenile hormone epoxide hydrolase through an expression host, Escherichia coli, to obtain a fermentation expression product of the juvenile hormone epoxide hydrolase.
[0028] (3) The present invention adds the recombinant protein of juvenile hormone epoxide hydrolase TjJHEH of Trichogramma oryzae to 10% honey water and feeds it to newly emerged Trichogramma oryzae adults, and characterizes that the recombinant protein of epoxide hydrolase TjJHEH can prolong the lifespan of Trichogramma oryzae adults. BRIEF DESCRIPTION OF THE DRAWINGS
[0029] The accompanying drawings are used to provide a further understanding of the present disclosure and constitute a part of the specification. Together with the following specific embodiments, they are used to explain the present disclosure but do not constitute a limitation of the present disclosure. In the accompanying drawings:
[0030] Figure 1 Schematic diagram of insect juvenile hormone epoxide hydrolase regulating insect juvenile hormone and growth and development;
[0031] Figure 2 It is a diagram of agarose gel detection of the juvenile hormone epoxide hydrolase TjJHEH gene of Trichogramma oryzae, PCR amplification of the juvenile hormone epoxide hydrolase TjJHEH gene of Trichogramma oryzae, lanes 1 and 2 are target fragments, and lane 3 is Maker;
[0032] Figure 3 is the relative expression level of the juvenile hormone epoxide hydrolase TjJHEH gene of Trichogrammatis oryzae 1-3 days after the emergence of Trichogrammatis oryzae adults; the expression level of 1-day-old Trichogrammatis oryzae adults was taken as 1, and different lowercase letters above the column indicate significant differences (P<0.05);
[0033] Figure 4 The figure is a SDS-PAGE detection diagram of the expression product of TjJHEH, a juvenile hormone epoxide hydrolase of Trichogramma oryzae, in E. coli; SDS-PAGE analysis of TjJHEH recombinant protein, the leftmost lane is the marker, lane 1 is the precipitate after pET28a-JHEH induced expression, lane 2 is the supernatant after pET28a-JHEH induced expression, lane 3 is the JHEH inclusion body protein, and lanes 4-5 are the purified JHEH proteins;
[0034] Figure 5 Graph showing the effect of TjJHEH recombinant protein of juvenile hormone epoxide hydrolase on the lifespan of T. oryzae adults. Effect of feeding TjJHEH recombinant protein on the lifespan of T. oryzae adults. Different lowercase letters above the columns indicate significant differences (P<0.05). DETAILED DESCRIPTION
[0035] The technical solution of the present invention is further described in detail below in conjunction with the accompanying drawings and specific embodiments. The present invention is further described below in conjunction with an implementation example. It should be noted that this embodiment is only used to explain the present invention, rather than to limit the scope of the present invention.
[0036] The invention discloses a recombinant protein of juvenile hormone epoxide hydrolase of Trichogramma japonicum, referred to as TjJHEH recombinant protein, wherein Tj is the abbreviation of the Latin name of Trichogramma japonicum, and JHEH is the abbreviation of juvenile hormone epoxide hydrolase in English, which has good application potential in the application of parasitic bees to control rice pests and reduce the amount of pesticides applied. Its amino acid sequence is shown in SEQ ID NO.1, and the nucleotide sequence encoding the recombinant protein is shown in SEQ ID NO.2. The invention relates to the field of genetic engineering, and uses an engineered strain of Escherichia coli as an expression host to achieve the expression of the recombinant protein of juvenile hormone epoxide hydrolase TjJHEH of Trichogramma japonicum. The invention identifies the juvenile hormone epoxide hydrolase of Trichogramma japonicum for the first time, and prepares a protein with biological activity. The preparation method of the recombinant protein is as follows: the coding sequence of the juvenile hormone epoxy hydrolase TjJHEH of the rice borer trichogrammatid is optimized by prokaryotic codons to obtain a nucleotide sequence, which is subjected to gene synthesis, and an expression plasmid is constructed and transformed into Escherichia coli BL21 (DE3). After induction expression and purification, the recombinant protein is obtained. The present invention also identifies the regulation of the juvenile hormone epoxy hydrolase on the life span of the adult trichogrammatid of the rice borer. The TjJHEH recombinant protein of the present invention can regulate the life span of the adult trichogrammatid of the rice borer, and can prolong the application time of the trichogrammatid of the rice borer. The obtained protein has important application potential in the use of rice borers to control rice pests.
[0037] Specifically, the invention obtains the juvenile hormone epoxide hydrolase TjJHEH gene of Trichogramma oryzae by PCR method, and heterologously expresses it by genetic engineering technology to obtain recombinant protein.
[0038] The present invention provides a recombinant vector comprising the juvenile hormone epoxide hydrolase.
[0039] The present invention provides a recombinant strain comprising the juvenile hormone epoxide hydrolase, and the recombinant strain is Escherichia coli pET28a.
[0040] The invention provides a quantitative detection method of the juvenile hormone epoxide hydrolase gene in insects.
[0041] The invention also provides application of juvenile hormone epoxide hydrolase in regulating the growth of adult Trichogrammatidae.
[0042] According to the technical solution of the present invention, the amino acid sequence of the recombinant protein of Trichogrammatus juvenile hormone epoxide hydrolase TjJHEH is shown in SEQ ID NO.1, and the Genbank accession number is XIF13460.1.
[0043] The present invention is achieved by the following technical means:
[0044] A recombinant protein of Trichogramma oryzae juvenile hormone epoxide hydrolase TjJHEH, the amino acid sequence of which is shown in SEQ ID NO.1.
[0045] First, the target fragment of juvenile hormone epoxide hydrolase of Trichogrammatus oryzae was amplified by PCR reaction, and the PCR fragment was obtained after agarose gel electrophoresis. The linearized fragment of the expression vector was prepared using a high-fidelity enzyme, and after agarose gel electrophoresis and product recovery and purification, it was homologously recombined with the prepared target PCR fragment and transformed into Escherichia coli DH5α competent cells by heat shock method to construct an Escherichia coli mutant library.
[0046] The screened positive transformants were all transferred into the expression host Escherichia coli BL21 (DE3) to perform a second round of screening of positive transformants. The screened transformants were shaken to obtain expression products, and after purification, the rice borer trichogrammatid juvenile hormone epoxide hydrolase TjJHEH recombinant protein provided by the present invention was screened. After verification by SDS-PAGE protein electrophoresis bands, the TjJHEH recombinant protein expression product was obtained.
[0047] Subsequently, the application function of the recombinant protein of the juvenile hormone epoxide hydrolase TjJHEH of the rice borer was analyzed. The recombinant protein TjJHEH was added to honey water to feed the newly emerged adults of the rice borer, and it was determined that the recombinant protein of the epoxide hydrolase TjJHEH could prolong the lifespan of the adults of the rice borer.
[0048] like Figure 2 As shown: the gene fragment of the Trichogrammatid juvenile hormone epoxide hydrolase TjJHEH recombinant protein of the rice borer of the present invention is obtained by screening through the PCR method.
[0049] The PCR method contains the following steps:
[0050] S1: The target fragment of juvenile hormone epoxide hydrolase was amplified by PCR reaction, and the primers used were: forward (5'-ATGGCGTTACTTTTGTTGACGG-3'), reverse (5'-CTACATCTCGGTTTTCTTCGAC-3'). The PCR fragment was obtained after agarose gel electrophoresis.
[0051] S2: Use high-fidelity enzyme to prepare linear fragment of expression vector, and then perform homologous recombination ligation with the prepared target PCR fragment after agarose gel electrophoresis inspection and product recovery and purification, and then transfer into competent E. coli DH5α by heat shock method to construct E. coli mutant library;
[0052] S3: All the screened positive transformants were transferred into the expression host Escherichia coli BL21 (DE3) to perform a second round of screening of positive transformants;
[0053] S4: The selected transformants were shaken to obtain expression products, the bacterial precipitate was resuspended in 20 mL of lysis solution, ultrasonically disrupted, and the ultrasonically disrupted cell lysate was centrifuged at 4°C 10000r / min for 20 min to collect the precipitate. The inclusion bodies were washed 3 times with inclusion body washing solution. The inclusion bodies were dissolved with a dissolution buffer, ultrasonically disrupted, and the supernatant was collected to obtain the provided rice borer trichogrammatid juvenile hormone epoxide hydrolase TjJHEH recombinant protein.
[0054] Experimental Materials and Reagents
[0055] 1. Strains and vectors:
[0056] The expression host Escherichia coli BL21 (DE3) was preserved in this laboratory; the expression vector pET28a was previously constructed and stored in this laboratory; and Escherichia coli DH5α was purchased from Invitrogen.
[0057] 2. Enzymes and kits:
[0058] PCR enzyme and ligase were purchased from Takara, and plasmid extraction kit and gel recovery kit were purchased from Sangon Biotechnology.
[0059] 3. Culture medium:
[0060] LB medium: 1% tryptone, 0.5% yeast extract, 1% NaCl;
[0061] Note: The molecular biology experimental methods not specifically described in the following examples are all carried out with reference to the specific methods listed in the book Molecular Cloning Experiment Guide, 3rd Edition by J. Sambrook, or according to the kits and product instructions.
[0062] Experimental methods
[0063] Example 1: Construction of juvenile hormone epoxide hydrolase TjJHEH engineering bacteria
[0064] (1) Determination of the amino acid sequence of the recombinant juvenile hormone epoxide hydrolase TjJHEH protein
[0065] The amino acid sequence of juvenile hormone epoxide hydrolase TjJHEH from Trichogramma oryzae published in GenBank (accession number: XIF13460.1, uploaded by the inventor) was used as the original amino acid sequence, and the amino acid sequence of the juvenile hormone epoxide hydrolase TjJHEH recombinant protein was obtained by PCR screening, as shown in SEQ ID NO.1.
[0066] (2) Construction of recombinant expression vector
[0067] The juvenile hormone epoxide hydrolase TjJHEH recombinant protein synthesis sequence was amplified by PCR reaction using PCR high-fidelity synthase. The PCR amplification program was 95℃ pre-denaturation for 5min, 95℃ denaturation for 30s, 50℃ annealing for 30s, 72℃ extension for 30s, a total of 35 cycles, and then 72℃ extension for 10min. The amplified fragment was recombined with the pET28a vector in vitro and transferred into the Escherichia coli competent cell DH5α, and the product was spread on LB solid medium and cultured at 37℃ for 12-16 hours until colonies grew. The grown single colonies were picked for positive clone verification, and the recombinant expression vectors after sequencing verification were preserved for subsequent experiments.
[0068] Example 2: Determination of the expression level of juvenile hormone epoxide hydrolase TjJHEH gene
[0069] (1) Obtaining the first-chain cDNA of Trichogrammatus oryzae. 100-2000 adult Trichogrammatus oryzae of the 1st, 2nd, and 3rd day were collected respectively, and RNA was extracted according to the instructions of the column-type animal tissue total RNA extraction and purification kit (Shanghai Sangon Biotech Co., Ltd.). The absorbance was detected by a spectrophotometer, and the quality of total RNA was detected by 1% agarose gel electrophoresis. The first-chain cDNA was synthesized from the extracted RNA using the reverse transcription kit, and the cDNA of each sample was stored in a -20℃ refrigerator for later use.
[0070] (2) Determination of the expression of the TjJHEH gene of Trichogrammatus oryzae. The expression of the TjJHEH gene of 1-3 day-old adult Trichogrammatus oryzae was determined using specific fluorescent quantitative PCR primers. The amplification program used was as follows: pre-denaturation at 94°C for 2 min; 94°C, 5 s, 60°C, 30 s, for 45 cycles; 95°C, 15 s, 60°C, 1 min (increase 0.3°C each time until 95°C); 95°C, 15 s; the reaction system was 20 μL: 0.4 μL of upstream and downstream primers, 10 μL of enzyme buffer, 1 μL of cDNA, and 8.2 μL of ddH2O. The primers used were: forward exf (5'-AGGGACGACGGCGGTTTAC-3'), reverse exr (5'-GCATCTTGTGGTTGAAGCTCTCC-3'), the internal reference gene was β-actin, the internal reference primers were forward exf (5'-AGTTGGTCCAGAATGTTCACTTGTC-3'), reverse exr (5'-CGTTGTGGTTGTCCCATTGAGG-3'). The real-time fluorescence quantitative PCR detector recorded the experimental data, with 2 ﹣ΔΔCTThe relative expression of the target gene was calculated by one-way analysis of variance. The significance of the data difference was compared by Tukey HSD multiple comparison method (P<0.05). The statistical analysis software was Excel 2013 and SPSS16. Figure 3 The results showed that the expression levels of the TjJHEH gene of Trichogrammatis oryzae were different in 1-3 day-old adult Trichogrammatis oryzae, indicating that the ecdysone regulatory protein TjERP of Trichogrammatis oryzae plays a role in the growth process of adults.
[0071] Example 3: Expression of juvenile hormone epoxide hydrolase TjJHEH recombinant protein in Escherichia coli
[0072] (1) Transformation of recombinant plasmid in Escherichia coli
[0073] Positive clones were transformed into BL21(DE3) E. coli strain.
[0074] (2) Shake flask expression of recombinant plasmids in Escherichia coli
[0075] The cells were cultured in LB medium at 30°C and 180 rpm for 12 h. The cells were broken, the bacteria were removed by centrifugation, the precipitate was collected, the inclusion bodies were washed 3 times with inclusion body washing solution, the inclusion bodies were dissolved with dissolution buffer, ultrasonically broken, and the supernatant was collected to obtain the rice borer trichogrammatid juvenile hormone epoxide hydrolase TjJHEH recombinant protein. The molecular weight band of the expression product was detected by SDS-PAGE, and the molecular weight was about 20 kDa. The results are as follows: Figure 4 shown.
[0076] Example 4: Juvenile hormone epoxide hydrolase TjJHEH recombinant protein regulates the growth of Trichogrammatidae adults
[0077] The experiment had a total of 4 treatments. The newly emerged Trichogrammatids of rice stem borer were taken and placed in glass tubes with stoppers, with 50 heads placed in each tube. A cotton ball soaked in 10% honey water was placed near the mouth of the glass tube and fixed with a stopper. TjJHEH recombinant protein was added to 10% honey water in the three treatments of the experimental group, with the final protein concentrations of 0.15μg / ml, 1.5μg / ml and 15μg / ml, respectively. No recombinant protein was added to the control group. Each treatment was repeated 3 times. The lifespan of Trichogrammatid adults in each treatment group was counted, and the mean value was calculated and analyzed by one-way analysis of variance. The significance of data differences was compared using the Tukey HSD multiple comparison method (P<0.05). The statistical analysis software was Excel 2013 and SPSS16. See Figure 5The results showed that the lifespan of T. oryzae adults fed with three concentrations of TjJHEH recombinant protein could be prolonged, which indicated that feeding TjERP recombinant protein could prolong the lifespan of T. oryzae adults by regulating and prolonging the application period, thereby improving the control effect of T. oryzae on rice pests.
[0078] The above is only a preferred embodiment of the present invention. It should be pointed out that due to the limitations of textual expression and the objective existence of infinite specific structures, ordinary technicians in this technical field can make several improvements, modifications or changes without departing from the principles of the present invention, and can also combine the above technical features in an appropriate manner; these improvements, modifications, changes or combinations, or the direct application of the inventive concept and technical solution to other occasions without improvement, should be regarded as the protection scope of the present invention.
Claims
1. A recombinant protein of juvenile hormone epoxide hydrolase of Trichogrammatus oryzae, characterized in that: The amino acid sequence of the recombinant protein is shown in SEQ ID NO:
1.
2. The recombinant protein of Trichogramma oryzae juvenile hormone epoxide hydrolase according to claim 1, characterized in that: The nucleotide sequence of the gene encoding the recombinant protein is shown in SEQ ID NO:
2.
3. The method for preparing the recombinant protein of Trichogramma oryzae juvenile hormone epoxide hydrolase according to claim 1 or 2, characterized in that: include: S1: The nucleic acid fragment of juvenile hormone epoxide hydrolase of Trichogrammatidae was amplified by PCR reaction, and the PCR fragment was obtained after agarose gel electrophoresis. S2: Use high-fidelity enzyme to prepare linear fragment of expression vector, and then perform homologous recombination ligation with the prepared target PCR fragment after agarose gel electrophoresis inspection and product recovery and purification, and then transfer into E. coli DH5α competent cells by heat shock method; S3: All the screened positive transformants are transferred into the expression host Escherichia coli to perform a second round of screening of positive transformants; S4: Inducing the expression of the engineered bacteria to obtain bacterial cells; S5: The bacterial cells are crushed, separated and purified to obtain the target protein, and the target protein is renatured to obtain the recombinant protein.
4. The method for preparing the recombinant protein of Trichogramma oryzae juvenile hormone epoxide hydrolase according to claim 3, characterized in that: The primers used in S1 were: forward 5′-ATGGCGTTACTTTTGTTGACGG-3′, reverse 5′-CTACATCTCGGTTTTCTTCGAC-3′.
5. The method for preparing the recombinant protein of Trichogramma oryzae juvenile hormone epoxide hydrolase according to claim 3 or 4, characterized in that: The PCR fragment in S1 was synthesized according to the nucleotide sequence shown in SEQ ID NO:
2.
6. The method for preparing the recombinant protein of Trichogramma oryzae juvenile hormone epoxide hydrolase according to claim 3 or 4, characterized in that: The expression vector in S2 is pET-28a, and the Escherichia coli in S3 is BL21 (DE3).
7. The method for preparing the recombinant protein of Trichogramma oryzae juvenile hormone epoxide hydrolase according to claim 3 or 4, characterized in that: In S4, the induction expression conditions are: at a bacterial solution OD 600 When the absorbance is 0.6-0.8, add 0.4mM-1.0mM IPTG and induce at 25-40℃ for 4-8h.
8. Use of the recombinant protein of juvenile hormone epoxide hydrolase of Trichogrammatus oryzae according to claim 1 or 2 for preparing a life-extending drug and / or a growth and development regulating drug for Trichogrammatus oryzae.
9. The use according to claim 8, characterized in that: The recombinant protein can extend the life span and / or regulate the growth and development of Trichogramma oryzae through feeding.
10. The use according to claim 8, characterized in that: The fluorescent quantitative detection method for the expression amount of the recombinant protein of Trichogrammatis juvenile hormone epoxide hydrolase in the adult of Trichogrammatis oryzae comprises the following steps: S1: RNA from the 1st, 2nd and 3rd day Trichogrammatidae adults was extracted separately and reverse transcribed to obtain cDNA; S2: The expression level of TjERP gene of 1-3 day old Trichogrammatidae adults was detected by fluorescence quantitative PCR reaction.