61 results about "Insect Gene" patented technology

The current invention provides methods to silence insect genes by using unpackaged dsRNA or siRNA, in one embodiment such dsRNA or siRNA is present in plant vascular tissue, preferably phloem, more particularly phloem sap, and the insect is a plant sap-sucking insect. Also provided are DNA sequences which when transcribed yield a double-stranded RNA molecule capable of reducing the expression of an essential gene of a plant sap-sucking insect, methods of using such DNA sequences and plants or plant cells transformed with such DNA sequences. Also provided is the use of cationic oligopeptides that facilitate the entry of dsRNA or siRNA molecules in insect cells, such as plant sap-sucking insect cells.

The invention provides a method for suppressing the gene expression of insects through feeding dsRNA, which belongs to the growth development field of adjusting and controlling insects through naturally feeding the dsRNA of the silenced and specific gene. The method selects a gene fragment SeA3 on chitin synthetase enzyme gene SeCHS-A, and studies the impact on the insects after the natural feeding. The method mainly comprises the steps: a proper expression vector L4440 is selected to construct a recombinant expression vector L4440-SeA3 with a purpose gene fragment SeA3; the recombinant vector is induced and expressed in specific recipient cell coliform bacteria HT115 (DE3) under vitro adaptive condition; and the recipient bacterium htLSA3 which expresses dsRNA is added into the feedstuff of the insects, the insect larvae are naturally fed with the feedstuff, the situation of the insects suppressing the gene expression is detected after the feeding, and the change of the phenotype of the insects is recorded. By adopting natural feeding to suppress the expression of the insect genes, the method explores new paths for controlling the insects.

Nanoparticles for insect RNAi via oral delivery are provided, along with methods of silencing a target gene in a target insect using RNAi are provided. The nanoparticles comprise a polymer matrix and insect dsRNA. The dsRNA comprises at least one sequence having a region of complementarity substantially complementary to at least a portion of an mRNA transcript of the target gene. Insect baits comprising the nanoparticles are also provided. Methods of screening target gene functions are also provided using the methods disclosed herein.

The invention discloses an artificial synthesized Bt insecticidal gene for transgenic anti-insect plants, further, the invention relates to the optimization and transformation systematically to a coding frame and a codon of a CrylAc anti-insect gene, according to an activity analysis to Bt protein in different regions and in accordance with the coding characteristics of the monocotyledons, obtaining anti-insect gene CrylAc-M which can be expressed in a monocotyledons efficiently, with the optimized codon and coding frame. The CrylAc-M gene after being transformed is connected on an expression carrier with CaMV35S-Adhl as promoter, and then the Bt gene CrylAc-Adhl is transferred into a maize inbred line with highly efficient conversion rate by a gene gun co-transformation method. A transgenic progeny is positive through Bt protein testing and a field anti-insect test of the field is tested to be highly resistant.

The invention discloses a pseudococcidae insect gene barcode detection technology which is capable of quickly detecting 21 types of pseudococcidae. The pseudococcidae insect gene barcode detection technology at least comprises a primer PseJ / PseN reagent which can be used over 20 times and a gene barcode sequence disk; a primer PseJ sequence is 5'-CCTTCAACTAATCATAAAAATATNAG-3', and a primer PseN sequence is 5'-TAAACTTCT GGATGTCCAAAAAATCA-3'; 93 entries of standard barcode sequence are stored on the gene barcode sequence disk, and detailed sequences are as shown in SEQ ID NO.1 to SEQ ID NO.93. The pseudococcidae insect gene barcode detection technology is a highly effective, accurate and convenient pseudococcidae insect molecular detection technology, and is capable of distinguishing among 21 types of pseudococcidae insects at the molecular level, thereby satisfying the use demands.

The present invention provides a genetic engineering material for insects that enables a target protein to be purified easily, without requiring the use of recombinant baculovirus, while simultaneously providing a process for producing exogenous protein using that genetic engineering material. A gene recombinant silkworm is obtained by inserting an exogenous protein gene such as a cytokine gene coupled to a promoter that functions in silk glands into a silkworm chromosome. An exogenous protein such as a cytokine is then extracted and purified from the silk glands or cocoon of that silkworm or its offspring. A large amount of exogenous protein can be produced within silk gland cells, outside silk gland cells or in silk thread or a cocoon by inserting an expression gene cassette, in which the DNA sequence of the 3′ terminal portion and the DNA sequence of the 5′ terminal portion of fibroin H chain gene are fused to the exogenous protein gene, into silk gland cells and so forth.

The invention clones insect specific neurotoxin gene BmkIT of East Asian scorpion and insect chitinase gene Chi to an insect cell-converting medium carrier pAcPDIEI to obtain a recombinant plasmid containing divalent anti-insect genes pAcPDIEI-Bmkit-Chi and transfecting the recombinant plasmid and DNA of AcNPV genome together in sfg cells, and obtaining a recombinant rhabdovirus containing divalent anti-insect genes AcNPV-BmKIT-Chi. The cooperation of the insect specific neurotoxin with the chitinase improves the poisoning effect of virus on cotton bollworms and other lepidopteral pests.

The invention relates to a synthetic root-specific promoter and application thereof, belonging to the technical field of biology. The synthetic root-specific promoter is named SRS1, and the nucleotide sequence is disclosed as SEQ ID NO 1; 35S promoter partial region and Omega sequence are added by an overlapping PCR process, and the synthetic sequence is SRSO1; by using pCAMB2300.1 as the basic vector (containing GUSplus report gene), the CaMV35S promoter on the pCAMB2300.1 is substituted by the synthetic promoter to obtain the expression vector p2300.1-SRSO1; and the promoter is transformed into agrobacterium, the verification on the promoter transferred into tobacco indicates that the promoter SRS1 has the root-specific expression characteristic. The synthetic promoter SRS1 enriches the varieties of the synthetic root-specific promoters, and can be used for root expression of plant anti-insect genes and for preventing and treating soil insects.

The invention provides a Xyleborus sp insect gene barcode detection kit. The Xyleborus sp insect gene barcode detection kit is used for rapidly detecting 12 kinds of Xyleborus sp, and comprises a primer reagent and a gene barcode sequence disk, wherein 37 standard barcode sequences are saved in the gene barcode sequence disk. The invention further discloses a method for performing Xyleborus sp insect molecular detection method by using the Xyleborus sp insect gene barcode detection kit. By adopting the kit and the method, convenient and efficient Xyleborus sp insect molecular detection can be performed, 12 kinds of Xyleborus sp insects can be distinguished on a molecular level, the use demand of quarantine identification can be met, and high practicability is achieved.

The invention provides a method for reinforcing plant-mediated insect RNA (Ribonucleic Acid) interference by using cysteine protease. In the method, a construct for expressing an insect gene dsRNA (double-stranded Ribonucleic Acid) and a construct for expressing plant cysteine protease are transferred to a plant cell, tissue or organ, so that the plant cysteine protease and the insect gene dsRNA can be expressed in the plant cell, tissue or organ.

The invention relates to the fields of nano-materials and gene interference, and concretely discloses a method for interfering the gene expression of an insect by infiltrating dsRNA through insect body walls. The adhesion, distribution and infiltration of exogenous dsRNA molecules to an insect body surface and entrance of the molecules to various internal organ tissues are realized by means of theeffect of a surfactant by complexing a nano-vector with a dsRNA functional macromolecule in order to efficiently interfere the gene expression. The efficient and noninvasive introduction of the exogenous dsRNA molecules to insect body cells by beans of the dual synergistic effects of the nano-vector and the surfactant in order to finally promote the correct targeting of the dsRNA molecules and improve the RNA interference rate. The method has the advantages of simplicity, high efficiency and small error, and is of great practical significance to the scientific research of the gene interference of the insect and the application of efficient and safe targeting RNA pesticide preparations.

The present invention pertains to an avidin-pseudotyped virus, and especially baculovirus, useful for delivery of foreign genes etc. The present invention also pertains to vectors comprising respective cassettes for pseudotyping, mammalian gene expression and insect gene expression in baculovirus.

The invention relates to the field of genetic engineering, and in particular relates to a cotton bollworm transport protein ABCC2 as well as a coding gene and an application thereof. The nucleotide sequence of the gene ABCC2 is shown in the SEQ ID No.2. In the invention, cotton bollworm transport protein ABCC2 is obtained through cloning, and the effect of a pesticide is enhanced by lowering the defense capability of target pests on biopesticide, so as to combine the biopesticide control with an insect gene silencing technology, open up a new field of pest control, and have important significance in lowering use amount of biopesticide, enhancing the pest control effect, improving food safety and increasing environment protection.

The invention provides a recombinant vector as well as a construction method and an application thereof, and belongs to the technical field of insect genetic engineering. The recombinant vector comprises a dual-luciferase reporter gene vector and a 3'UTR region nucleotide sequence of a nilaparvata lugens Ccdc 124 gene. The construction method of the recombinant vector comprises the following stepsof 1) performing PCR amplification to obtain the 3'UTR region nucleotide sequence of the nilaparvata lugens Ccdc 124 gene by taking the cNDA of nilaparvata lugens as a template; and 2) recombining the 3'UTR region nucleotide sequence of the nilaparvata lugens Ccdc 124 gene into the dual-luciferase reporter gene carrier to obtain the recombinant vector. The recombinant vector provided by the invention can determine the expression activity of the nilaparvata lugens Ccdc 124 gene, is used for screening microRNA for regulating the nilaparvata lugens Ccdc 124 gene, and can also study the regulation mechanism of the predicted microRNA regulated Ccdc 124 gene.

The invention discloses a MeJA inducement promoter separated from mustard that could construct recombination expression carrier using the connecting of the external source gene. When the external gene is anti-insect gene, the transformed plant would be gained.

The invention relates to the technical field of insect gene engineering and particularly relates to an isolated adelphocoris suturalis Taiman gene and a protein encoded by the same. The gene has a nucleotide sequence as shown in SEQ ID No:1. The dsRNA of the Taiman gene is injected into a newly emerged female insect body of adelphocoris suturalis through microscope, and detection discovers that the expression amount of the Taiman gene is extremely remarkably inhibited in the whole producing period, the number of eggs in the ovary of adelphocoris suturalis and the lifetime egg laying amount ofadelphocoris suturalis are remarkably reduced by inhibiting the expression of the Taiman gene, and the reproductive capacity and population development of the adelphocoris suturalis are influenced. The Taiman gene can be used as a backup gene for culturing anti-adelphocoris suturalis transgenic plants.

The invention discloses a method for suppressing insect gene expression by short single-stranded DNA. The method comprises the following steps: 1) determining an insect target gene, performing PCR (polymerase chain reaction) amplification to obtain a DNA sequence of an insect target gene, and performing sequencing; 2) designing a short single-stranded DNA fragment according to the DNA sequence obtained in step 1); 3) diluting the short single-stranded DNA fragment obtained in step 2), and injecting the short single-stranded DNA fragment into an insect body by microinjection; 4) performing RT-qPCR detection on expression of the insect target gene in step 3) to determine the relative expression level of the inset target gene.

The present invention provides a chilo suppressalis SDR gene and an encoded protein and an application thereof, dsRNA and an amplified primer pair and an application thereof, and belongs to the technical field of insect genetic engineering. The chilo suppressalis SDR gene has a nucleotide sequence shown in SEQ ID No.1. The provided chilo suppressalis SDR gene participates in growth and developmentprocesses of chilo suppressalis, deletion of the gene significantly increases mortality of larvae of the chilo suppressalis. The dsRNA of the chilo suppressalis SDR gene inhibits expression of the chilo suppressalis SDR gene.

The invention provides a recombinant vector and its construction method and application, which belong to the technical field of insect genetic engineering. The recombinant vector includes a dual-luciferase reporter gene carrier and the nucleotide sequence of the 3'UTR region of the brown planthopper Ccdc124 gene; The method for constructing the recombinant vector comprises the following steps: 1) using the cDNA of the brown planthopper as a template, performing PCR amplification to obtain the 3'UTR region nucleotide sequence of the Ccdc124 gene of the brown planthopper; 2) converting the 3'UTR of the Ccdc124 gene of the brown planthopper to The nucleotide sequence of the region was recombined into the dual luciferase reporter gene vector to obtain a recombinant vector. The recombinant vector provided by the invention can measure the expression activity of the Ccdc124 gene of the brown planthopper, be used for screening the microRNA that regulates the Ccdc124 gene of the brown planthopper, and can also study the regulation mechanism of the predicted microRNA regulating the Ccdc124 gene.

The invention provides a lepidoptera insect Hpx12 gene and application. The first purpose of the invention is to provide a lepidoptera insect innate immune response regulation gene Hpx12; the second purpose is to provide a cloning method of the lepidoptera insect innate immune response regulation gene Hpx12; and the third purpose is to provide the application of the lepidoptera insect innate immune response regulation gene Hpx12. According to the invention, after the expression of the lepidoptera insect Hpx12 gene is inhibited, the innate immune response of insect bodies can be inhibited, the susceptibility of the insect bodies to germs is increased, silkworms are more easily infected with BmNPV viruses, and the death rate of lepidoptera insects is further greatly increased. If the expression of lepidoptera insects Hpx12 is inhibited, the purpose of pest control can be achieved. The lepidoptera insect Hpx12 gene has positive significance in solving the problems of ecological deterioration, insecticide resistance and the like caused by chemical insecticides.

The invention provides a trypsin precursor gene and its encoded protein, interfering RNA and an application thereof, which belong to the technical field of insect gene engineering. A cDNA sequence ofthe trypsin precursor gene is shown as SEQ ID NO: 1; an amino acid sequence of the protein is shown as SEQ ID NO: 2; a nucleotide sequence of the interfering RNA is shown as SEQ ID NO: 3; the expression level of the trypsin precursor gene in adelphocoris suturalis in Miridae is reduced, the quantity of eggs in the adelphocoris suturalis ovary and the final production of eggs can be significantly reduced, and the life span of female adults can be shortened. The application provided by the invention can eventually lead to the decline of the population development, provides a new idea for controlling the development of the adelphocoris suturalis population, and also provides a theoretical basis for the realization of adelphocoris suturalis and other hemipteran insects in green control.

The invention provides a method for noninvasive identification on a genotype of an insect. The method comprises the following steps: (a) separately feeding wild type insects and gene-edited insect mutants, and independently marking or feeding each insect individual; (b) employing that each ecdysis of each insect is regarded as once increase of age, and separately collecting ecdysis of corresponding insects from 1 age to an adult stage; (c) extracting genomic DNA from the ecdysis by using a DNA extraction kit; and (d) taking the genomic DNA extracted from the ecdysis as a template, designing primers according to a target gene sequence, carrying out PCR amplification, sequencing an amplification product, and then, identifying the genotype. According to the method, noninvasive, early-stage and repeated insect genotype identification can be achieved by using the ecdysis, and the problems that mechanical injury is caused by scissoring insect body tissue and insect bodies of a juvenile stagecannot be identified are solved.

The invention relates to a gene extraction device, in particular to a device for extracting genes of flying insects. The technical problem to be solved is to provide the device for extracting the genes of the flying insects. The device for extracting the genes of the flying insects comprises a rectangular frame, an operating screen, a first vertical plate, a first cross plate, a second cross plate, a left wing separation mechanism, a right wing separation mechanism and a DNA separation and extraction mechanism, wherein the operating screen is arranged at the left end of the rectangular frame;the left side of the top end of the rectangular frame is connected with the first vertical plate; the left wing separation mechanism is arranged on the inner left side of the rectangular frame; and the right wing separation mechanism is arranged on the inner right side of the rectangular frame. According to the device disclosed by the invention, completeness of wings of the insects is ensured, thewings are completely separated from the insect bodies, and the heads of the insect bodies are completely separated from active cells of the tail parts, so that the completeness of internal structuresof the insect bodies is ensured, and DNA is effectively separated from cell-attached proteins and cell organelles, and thus, a large quantity of accurate gene groups can be obtained, and experimentalaccidental effects are excluded.

An insect gene drive system for biocontrol of a population of an insect is provided. The gene drive system includes: a) a first DNA sequence encoding a toxin under the control of a maternal germline-specific promoter active in the insect, with the first DNA sequence being linked to b) a second DNA sequence encoding an antidote under the control of an early embryo-specific promoter active in the insect. The toxin is expressed in maternal germline cells of the insect and results in maternal-effect lethality in the insect, and the antidote is expressed in embryos of the insect and counters the maternal-effect lethality. In some embodiments, the insect is Drosophila suzrukii.

The invention belongs to the field of reporter gene vectors, and particularly relates to a dual-luciferase reporter gene vector for insect cells, a construction method, a recombinant vector and application. The dual luciferase reporter gene vector for the insect cells is obtained by modifying a psiCHECK-2 vector; the transformation comprises the following steps: replacing an original SV40 promoter with an OpIE2 promoter, and inserting a synthesized DNA (Deoxyribose Nucleic Acid) fragment between an HSV-TK promoter and a Synthetic poly (A); the downstream of the hRluc gene of the psiCHECK-2 vector is used for inserting an insect target gene of interest; the nucleotide sequence of the synthesized DNA fragment is as shown in SEQ ID NO: 1. The dual-luciferase reporter gene vector for insect cells provided by the invention ensures powerful gene transcription and mRNA processing in insect cells, and is suitable for insect gene function research.

The invention discloses an artificially synthesized anti-insect protein mCry1Ia2. The invention further discloses a nucleic acid molecule of the protein, a recombinant vector, a host cell or a recombinant bacterium of the nucleic acid molecule. The invention further discloses application of the protein as well as the nucleic acid molecule, the recombinant vector, the host cell or the recombinant bacterium of the nucleic acid molecule, and the like in regulating and controlling anti-insect properties and insect killing performance of plants or improving the anti-insect properties of transgenicplants, and preparing insecticides or anti-insect preparations. In-vitro experiment shows that the transformed and synthesized Bt gene toxin production protein has a remarkable insect killing effect on ostrinia nubilalis. A constitutive promoter is adopted to construct a novel plant expression vector, an anti-insect gene mCry1Ia2 can be stably and efficiently expressed in monocotyledons, and furthermore, anti-insect transgenic plants can be produced.