230 results about "Transgenic lines" patented technology

Transgenic plants are more tolerant to environmental stresses than untransformed plants. The pea ABR17 (Abscisic acid responsive 17) is used to enhance germination of plants such as Arabidopsis sp. and Brassica sp. while under multiple abiotic stresses, and to enhance the tolerance of these plants to these stresses. Three independently derived Arabidopsis transgenic lines, containing ABR17, germinated better in the presence of salt, cold temperature or both. The transgenic plants also exhibited enhanced tolerance to freezing temperature or extreme heat. Furthermore, the transgenic plants demonstrated early flowering even under normal, non-stressed conditions.

The invention discloses a method for creating rice engineering maintainer lines preventive against gene flow. The method includes the steps of 1, site-specifically inserting an exogenous reporter gene and a pollen lethal gene into a flanking sequence of a fertility control gene of a wild rice chromosome so as to obtain a transgenic line containing the exogenous reporter gene and the pollen lethal gene, wherein the exogenous reporter gene, the pollen lethal gene and the fertility control gene are able to closely link and inherit; 2, hybridizing wild rice and homozygous recessive rice common genic male sterile lines to obtain F1-generation plants, hybridizing the F1-generation plants as male parents to the transgenic line to obtain a generation F2, screening the generation F2 to obtain hybrid lines containing the exogenous reporter gene, namely rice engineering maintainer lines required, wherein the homozygous recessive rice common genic male sterile lines are mutant lines with the sterility control gene. The invention further discloses application of the rice engineering maintainer lines preventive against gene flow in rice breeding.

A method of expressing in vivo heart-specific fluorescence in transgenic line of zebrafish is developed, which provides a research model for studying heart-related gene functions and performing gene therapies in the future. The method comprises the following steps. Firstly, a plasmid is constructed. This plasmid construct includes the upstream regulatory region, the exon 1, the intron 1, and the exon 2 of cmlc2 gene, cDNA of GFP, wherein the cmlc2 gene and GFP cDNA form a cassette, and inverted terminal repeats from adeno-associated virus are flanked at both sides of this cassette. The plasmid construct is linearized and microinjected into one-celled zebrafish fertilized eggs. Lastly, the heart-specific fluorescent expressed zebrafish are selected and the germline-transmitting transgenic strain is generated.

A method of expressing in vivo heart-specific fluorescence in transgenic line of zebrafish is developed, which provides a research model for studying heart-related gene functions and performing gene therapies in the future. The method comprises the following steps. Firstly, a plasmid is constructed. This plasmid construct includes the upstream regulatory region, the exon 1, the intron 1, and the exon 2 of cmlc2 gene, cDNA of GFP, wherein the cmlc2 gene and GFP cDNA form a cassette, and inverted terminal repeats from adeno-associated virus are flanked at both sides of this cassette. The plasmid construct is linearized and microinjected into one-celled zebrafish fertilized eggs. Lastly, the heart-specific fluorescent expressed zebrafish are selected and the germline-transmitting transgenic strain is generated.

The present invention relates to the mitogen-activated protein kinase called MAPK5. The rice MAPK5 gene, its protein and kinase activity were induced by abscisic acid, pathogen infection, wounding, drought, salt and cold temperature. However, suppression of MAPK5 expression and kinase activity in dsRNAi transgenic plants resulted in constitutive expression of pathogenesis-related genes such as PR-1 and PR-10 but enhanced resistance to fungal and bacterial pathogens. In contrast, overexpressed transgenic lines exhibited elevated MAPK5 kinase activity and increased tolerance to drought, salt and cold stresses. This invention provides methods for increasing tolerance to abiotic and biotic stress in plant using MAPK5.

The present invention relates to a transgenic Trichoderma spp. having a recombinant nucleic acid molecule encoding a bioactive molecule. The present invention also relates to methods for controlling plant disease that involve applying a transgenic strain of Trichoderma spp. plants or plants seeds, where the transgenic strain of Trichoderma spp. has a recombinant nucleic acid molecule encoding a bioactive molecule that is capable of controlling plant disease and conferring systemic disease resistance to the plant or a plant grown from the plant seed. The present invention also relates to a method of delivering a bioactive molecule to a plant or plant seed. The method involves providing a transgenic strain of Trichoderma spp. having a recombinant nucleic acid molecule encoding a bioactive molecule and applying the transgenic strain of Trichoderma ssp. the plant's roots or seeds.

This invention provides a method for the development of novel plant germplasm using segregates from a transgenic line combined with PCR-based zygosity testing and, optionally, Southern blot analysis.

The invention discloses a vector for efficiently labeling zebra fish PGC, and a preparation method and a use of transgenic fish. A Gal4 / UAS transcription activation system is utilized and mRFP is used as a report gene to realize an inductive gene expression regulation technology in the primordial germ cells of zebra fish. The preparation method of the transgenic fish comprises the following steps: 1, respectively constructing activation transgenic line Tg (kop:KalTA4) and an effect transgenic line Tg (UAS:mRFP); and 2, hybridizing Tg (kop:KalTA4) male fish with Tg (UAS:mRFP) female fish to obtain the transgenic fish for efficiently labeling the primordial germ cells of zebra fish. The transgenic fish for efficiently labeling the primordial germ cells of zebra fish can be widely applied to the fish bioengineering.

The invention discloses a method for determining an insertion site of a transgenic line by a re-sequencing technique. The method comprises the following steps of (1) extracting DNA (deoxyribonucleic acid) of a genome of the transgenic line, wherein the total amount is at least 2g; (2) re-sequencing the genome, wherein the data size is at least 12G; (3) using a T-DNA sequence of a transgenic carrier as a template, and comparing and analyzing the obtained sequencing data, so as to obtain information of the insertion site; (4) according to the information of the insertion site, designing a primer, and performing PCR (polymerase chain reaction) verification, so as to determine the insertion site of each transgenic line. The method for determining the insertion site of the transgenic line has the characteristics of maturity, simplicity and high success rate, and can be widely applied to determine the insertion site of the transgenic line.

The present invention relates to a construction method of a human dcf1 gene transgenic drosophila melanogaster model. According to the invention, a drosophila melanogaster transgenic vector pUAST-dcf1 is constructed; a human dcf1 transgenic UAS drosophila melanogaster strain is obtained through microinjection; and human dcf1 transgenic drosophila melanogaster strains, with balance gene and genetic stability, of w, UAS-dcf1 / Cyo and Tm6B / Tm2 are obtained through hybrid and backcross of the human dcf1 transgenic UAS drosophila melanogaster strain with Double Balance drosophila melanogasters of w, B1 / Cyo and Tm6B / Tm2. The present invention utilizes microinjection gene engineering technology and drosophila melanogaster hybridization technology to construct the human dcf1 gene transgenic drosophila melanogaster model, and provides materials and new ideas for research on relative neurodegenerative diseases.

The present invention provides Maternal Sterility Constructs (MSC) and methods of producing sterile progeny lacking germ cells. Female animals carrying the MSC transgene will give rise to a sterile generation, as the MSC specifically eliminates Progenitor Germ Cells (PGCs) of her progeny. These females are called lineage ending females. Male animals carrying the MSC transgene, however, give rise to fertile progeny (assuming the male is not derived from an MSC-transgenic female). Thus, MSC transgenic males can be used to propagate the transgenic line. The invention can be advantageously applied to eliminate pest or invasive species, or to provide effective population control and improve culture performance of farmed species, such as fish and shellfish.

The invention discloses an application of an OsGRF6 protein in regulation of plant height. According to the application of the OsGRF6 protein in the regulation of the plant height provided by the invention, an amino acid sequence of the OsGRF6 protein is sequence 2 in a sequence table. In the above application, a nucleotide sequence of a coding gene of the OsGRF6 protein is sequence 1 in a sequence table. Experiments show that a gene OsGRF6 of GRF family is cloned in japonica rice (ZH10); and positive transgenic rice lines are obtained by transferring rice ZH10 callus by an agrobacterium-mediated method. By analyzing the transgenic lines and corresponding wild type phenotypes, obvious dwarf characteristics are found in the transgenic rice.

The invention discloses a plant inducible promoter and an application thereof. A section of HbCIPK2 promoter sequence of 1,750bp is obtained from a halophyte wild barley genome through a chromosome primer walking method, an arabidopsis proHbCIPK2:GUS strain obtained through the promoter sequence can drive expression of GUS genes under stress of different concentrations and times, and the expression activity of GUS is enhanced along with increase of the stress times and concentrations. The inducible expression activity of the GUS genes driven by the HbCIPK2 promoter under PEG and ABA stress is relatively low. The identical transgenic lines are free of GUS gene expressions under stress-free treatment and low-temperature and high-temperature stress. The obtained DNA sequence of 1,750 bp is the novel stress inducible promoter, and has high promoter activity under high-salt stress, and proper expressions of transcription factors and other controlling genes can be induced.

Disclosed is a rice waterlogging stress response RS1 gene. The rice waterlogging stress response RS1 gene is characterized in that a DNA sequence of the RS1 gene is shown as SEQ ID NO:1 in a sequence table, and an amino acid sequence table of coded protein of the rice waterlogging stress response RS1 gene is shown as SEQ ID NO:2. The rice waterlogging stress response RS1 gene has the advantages that a ubi promoter is selected to drive the RS1 gene, so that waterlogging tolerance of rice is improved remarkably by an over-expressed transgenic line, waterlogging tolerant effect is obvious, and in a waterlogging environment, the RS1 gene can obviously improve waterlogging tolerance of the rice and guarantee high yield and stable yield of the rice; the RS1 gene is from the rice, so that worry, of people, about food safety is reduced by the rice with the RS1 genetically modified; a molecule mechanism for forming waterlogging tolerance of the rice is revealed, so that a new gene resource is provided for genetic improvement on waterlogging tolerance of the rice and other plants.

The invention provides a gene for controlling a rice stem-leaf angle and application of the gene. The gene CYC U4;1(Os10g41430) disclosed by the invention specifically expresses in a rice pulvinus (believe that the gene also specifically expresses in other graminaceous crops), the CDS sequence of the gene is shown in the SEQ ID No.2, and a coded amino acid sequence is shown in the SEQ ID No.3. The invention further provides a promoter sequence, shown in the SEQ ID No.1, for the gene. A promoter of CYC U4;1 and overall length CDS are cloned to pCAMBIA1301, and then converted into Nipponbare, leaf-stem angles of the obtained transgenic lines are less than those of wild rice, therefore, plant type growth of plants can be controlled by increasing or decreasing expression quantity of CYC U4;1 gene through a genetic engineering technology, plant types are improved and germplasm density is increased.

The invention relates to a construction method and application of a humanized mouse model, in particular to a construction method and application of a transgenic mouse model of a human Nbs1<c.657del5> gene. The method includes: constructing a 5bp deletion mutation-containing BAC carrier in the human Nbs1 gene, conducting pronuclei microinjection, and performing screening to obtain 3 stable transgenic lines for high expression and low expression of the human Nbs1 gene. The transgenic mouse involved in the invention has the phenotypes of delayed puberty, uniform shortening of body length and bone dysplasia at certain proportion in one of the lines, and a new mouse model is established for Nijmegen breakage syndrome diseases. At the same time, as the Nbs1 gene function impairment is closely related to cancers, the transgenic model can be applied to short-term carcinogenic tests in drug pre-clinical safety evaluation, thus providing a potential substitution model for traditional biennium carcinogenic tests and also providing an effective tool for research of carcinogenesis mechanisms.

A disclosed preparation method of transgenetic zebrafish used for detecting environment pollutants comprises: (1) cloning the promoter regulatory sequence of Mdr1 protein from zebrafish genome; (2) fusing the DNA fragment obtained by cloning into the upstream of GFP coding sequence for constructing an expression vector; (3) performing zebrafish embryo microinjection on the expression vector; (4) culturing the embryo subjected to injection to sexual maturity, and enabling cultured zebrafish and wide zebrafish to copulate and oviposit; and (5) screening positive F1 generation under a fluorescence microscope, and performing inbreeding to obtain F2 generation and further to obtain a transgenic line with stable inheritance. Because the expression level of fluorescin can be induced in a concentration dependent way by multiple environment pollutants such as heavy metals, polycyclic aromatic hydrocarbons and the like, the transgenic zebrafish prepared by employing the preparation method can be used to detecting the existence and the concentration of the above chemical pollutants in environment; and the preparation method has the advantages of being economic, rapid, convenient and accurate.

The invention discloses application of a transcription factor involved in regulating tomato steroid alkaloid synthesis. The amino acid sequence of the transcription factor SlbHLH114 is as shown in SEQID NO.4, and the nucleotide sequence is as shown in SEQ ID NO.3. Overexpression of the transcription factor has a positive regulating effect on the synthesis of tomato steroid alkaloids, so the steroid alkaloid content of transgenic lines can be improved by overexpressing the transcription factor SlbHLH114 in plants, and a theoretical basis is provided for tomato quality improving and excellent breeding.

The invention relates to an exogenous insert vector flanking sequence for transgenic rice and application of the flanking sequence and belongs to the technical field of biology. According to the invention, a flanking sequence of an exogenous insert fragment of a transgenic rice variety SK-2 is cloned for the first time; the application that the flanking sequence of the exogenous insert fragment of the transgenic rice variety SK-2 can be used as a target DNA (Deoxyribose Nucleic Acid) amplification fragment is defined; and the variety specific qualitative PCR (Polymerase Chain Reaction) detection of a specific transgenic rice variety SK-2 is established. The cloned and separated flanking sequence of the exogenous insert fragment can be further used for establishing a transgenic rice variety SK-2 PCR detection method. The flanking sequence can be used for designing a specific primer for the amplification of target DNA and establishing the variety specific qualitative and quantitative PCR detection of the specific transgenic rice variety SK-2.

The invention relates to molecular biology, and particularly discloses a method for detection of transgenic crops.Inner primers and outer primers are designed according to the endogenous gene, exogenous gene and transgenic strain flanking sequence of a transgenic crop, all the inner primers and outer primers are mixed firstly for the first round of high-pass multi-enrichment quantitative PCR with a small cycle number (15 cycles), then the second round of fluorogenic quantitative PCR is conducted with the inner primers, a result is analyzed according to an amplification curve and a fusion curve, and in this way, the success rate and stability of the method are improved remarkably and the purpose of high-pass detection of transgenic crops can be realized.The sensitivity of the method is higher than that of ordinary fluorogenic quantitative PCR by about one magnitude order; the method has the same quantitative capacity as ordinary fluorogenic quantitative PCR, has high specificity and is a more effective method for detection of transgenic crops.

Provided is an application of an OsmiR156f gene in rice effective tillering increasing. The OsmiR156f gene which has important regulating effect on rice tillering development is selected from a miR156 family, a precursor pri-miR156f sequence is cloned, through a stem specific promoter vector construction method, the OsmiR156f gene which can increase tillering is guided into rice, specific expression in the stem of rice is carried out, and rice tillering development is regulated. The early tillering capacity of transgenic rice can be improved properly, tillering sprouts at a high section and later-period tillering are still restrained, accordingly, the effective tillering quantity of a single rice plant can be increased, ineffective tillering quantity is not increased, a transgenic line rice ear number is increased properly, influence on yield correlated characters such as rice ear length and thousand kernel weight is small, and finally the purpose of single-plant production increasing is achieved.

The invention relates to a rice flowering phase control protein and application thereof in breeding. An amino acid sequence of an OsLFT1 protein, an OsLFT1 nucleotide sequence encoding the OsLFT1 protein and a nucleotide sequence of an OsLFT1 gene promoter are included. For the first time, the rice OsLFT1 gene is cloned, an OsLFT1 overexpression plant expression vector is constructed, the OsLFT1 promoter starts the plant expression vector of a GUS report gene, OsLFT1 overexpression is created, the OsLFT1 promoter starts a rice transgenic line of the GUS report gene, a dwarf and later-floweringrice breeding material is obtained, and the function of OsLFT1 in rice is researched deeply. According to the cloned OsLFT1 gene and the created dwarf rice breeding material with the changing flowering stage, the rice anti-lodging capability is improved in rice dwarfing breeding; for rice variety breeding in different growing periods, the problem that the hybridized rice flowering stage is sterile is solved, and the protein has the wide application prospect in the aspects of increasing the rice yield and improving the rice quality.

A process for configuring a transgenic line of early-mature grass includes such steps as sampling the stem tip of early-mature seedling, in vitro culturing to induce its gibbosity, taking its tissue, culturing to induce its bunch sprouts, reproducing the bunch sprouts rooting culture to obtain complete plant, and using gene gun to bombard the sprout tuber to obtain its transgenic plant.

The invention discloses a marker set and composition for comprehensively screening genetically modified ingredients and application. The marker set comprises seven screening elements including PCAMV35S, TCAMV35S, TNOS, PRbcS4, TPINII, TE9 and PAT; the marker set further comprises strain-specific sequences of corn DAS40278, a soybean DP305423 and a soybean CV127. The composition is a reagent for detecting the marker set, preferably primers and probes shown in SEQ ID No.1-33. The marker set and composition can cover 66 commercial genetically modified strains on the market, have the advantages ofgood repeatability, high throughput, sensitivity, accuracy and rapidity, and have good application prospects in the aspect of detecting the genetically modified components.

The invention discloses a cotton drought resistance related gene GhDT1 and an application thereof, and belongs to the field of biotechnology application. The invention relates to the gene GhDT1 whichencodes a lipid transfer protein. The invention provides a full-length ORF nucleotide sequence and an amino acid sequence of GhDT1 in an allotetraploid upland cotton genetic standard line TM-1. The gene GhDT1 in the cotton undergoes dominant expression in fiber tissues and is subjected to abiotic stress and hormone-induced expression. Overexpressed and interference expressed GhDT1 transgenic cotton materials are obtained through an agrobacterium mediated genetic transformation method. Through identification of natural drought in the seedling stage of a progeny homozygous transgenic line, it isfound that compared with a transgenic receptor material, a plant containing the overexpressed gene has higher drought tolerance, drought tolerance is remarkably reduced through interference expression, and it is indicated that the gene plays an important role in the drought stress resisting process of cotton.

The subject invention provides methods of selectively recognizing and / or eliminating potentially undesirable soybeans, from the soybean supply and / or selectively preserving specific soybean seed identities comprising: the genetic transformation of small variety soybeans; the separation and isolation of said soybeans; and, optionally, the replanting and harvesting of the soybeans, thereby resulting in stable transgenic lines.

The invention provides a wheat TaARF12 gene and application thereof, and belongs to the field of molecular biology of crops. A designed primer clones a coding region sequence of TaARF12 from cDNA of Chinese spring young ears, and the coding region sequence of the TaARF12 gene in homologous chromosomes A, B and D is represented as SEQ ID NO. 1-3 respectively. A TaARF12-specific RNAi fragment is selected, an RNAi vector is created, and Fierder is transformed. A CRISPR / Cas9 vector is used to create a gene editing vector containing two target sites and transform wild-type Fierder. Obtained RANi transgenic lines and gene editing lines have similar phenotypes, with plants becoming shorter and ears becoming longer. It is indicated that the wheat TaARF12 gene can regulate the plant height and theear length, and the plants become shorter and the ears become longer after the gene is interfered and expressed.

The invention belongs to the field of biotechnology, and discloses a method for improving the vitamin C content of a tomato quality component by multi-gene polymerization. According to the method, keystructural genes S1GMP, SlGME, SlGGP and SlGPP are selected in the pathway of polymerizing D-Man / L-Gal by hetero-binding labeling; the AsA content and AsA excretion content in petioles and stalks ofdifferent polymerized transgenic lines are determined; and the increment on the AsA content, AsA transport capacity and tolerance to oxidative stress of tomatoes through gene polymerization are analyzed. By cross-polymerizing the overexpression transgenic lines of the four key structural genes in the ascorbic acid synthesis pathway, namely D-mannose / L-galactose pathway, so that the ascorbic acid content and transport in tomato fruits and leaves are increased in polymerized plants, the tomato resistance to oxidative stress is improved simultaneously, and a novel method is provided for cultivation of high-quality tomatoes and resistance breeding.

The invention discloses a construction method of a rice engineering maintainer line. The method comprises the following steps: 1, integrating an exogenous reporter gene to the chromosome of wild type rice in a site-specific integration manner to obtain a transgenic line containing the exogenous reporter gene, and then executing the step 2, wherein the exogenous reporter gene and a wild type fertility regulatory gene can be subjected to close linkage inheritance; 2, hybridizing the transgenic line obtained in the step 1 with a homozygous recessive rice common nuclear male sterile line, and screening a strain containing the exogenous reporter gene as the needed rice engineering maintainer line, wherein the homozygous recessive rice common nuclear male sterile line is a fertility regulatory gene mutant strain. The invention further discloses application of the rice engineering maintainer line to rice breeding. By integrating the exogenous reporter gene to the chromosome of the wild type rice in the site-specific integration manner, the problem of limitation on transformation of a receptor material is solved. The rice engineering maintainer line is available for rice common recessive nuclear male sterile line propagation.

The invention discloses a method for identifying heat resistance of capsicum by the utilization of in vitro leaf disk. The method mainly includes capsicum heatproof line B35 used as a heatproof contrast, capsicum heat-labile line B6 used as a temperature-sensitive contrast, heat resistance identification of capsicum in vitro leaf disk, heat damage grading standard of in vitro leaf disk and the like. The method is characterized in that when the capsicum in vitro leaf disk is processed at specific temperature (high temperature and normal temperature), at specific concentration of a salt solution and within a certain period of time, different heatproof lines have distinct differences in leaf disk phenotype symptom. On this account, through statistics and calculation of heat damage index of some line and according to the established heat damage grading standard, heat resistance lines and heat sensitivity lines of capsicum can be identified. The method is especially suitable for identification and genetic analysis of heat resistance of those rare and transgenic lines of capsicum material, is a method which is simple and rapid to operate and effectively identifies heat resistance of the capsicum germplasm resources, and can be adopted to accelerate the progress of capsicum heatproof breeding.