15 results about "Target gene" patented technology

The invention discloses a genetic transformation method by the utilization of cotton meristematic tissue in the plant gene engineering field. According to the method, without the step of cotton tissue culturing, activated Agrobacterium containing target gene plasmid vector is directly dipped on a growing point removed cotton plant to induce adventitious bud to grow until the adventitious bud blossoms and grows bolls, and transgenic cells are screened by the use of antibiotics. By the utilization of the method for genetic transformation of cotton, differentiation rate is high, the screening method is simple, the cost for cotton genetic transformation is saved, and the time for cotton genetic transformation is shortened.

The invention discloses a novel gene cloning method independent of enzymes. The novel gene cloning method comprises the following steps of amplifying a target gene and a plasmid vector with a primer;performing heat treatment on target gene amplification products and plasmid vector amplification products with iodine; performing hybridization pairing on target gene DNA fragments and plasmid vectorDNA fragments after heat treatment, to form a recombinant plasmid with a notch, containing the target gene; and transforming escherichia coli with the recombinant plasmid with a notch, containing thetarget gene, and repairing the notch of the recombinant plasmid to form a complete recombinant plasmid containing the target gene. In the manner, the best sulfo modifying and single-strand end lengthcontrol strategy is adopted, the positive cloning rate of the target gene is high, the operation flux is high, and the method is suitable for simultaneous cloning of a plurality of target genes.

The invention belongs to the technical field of biology and in particular relates to an absolute fluorescence quantitative PCR (Polymerase Chain Reaction) primer and a kit for detecting an atypical porcine pestivirus (APPV). Nucleotide sequences of the absolute fluorescence quantitative PCR primer for detecting the atypical porcine pestivirus are shown as SEQ ID NO. 1 and SEQ ID NO. 2; the primer can be used for specifically amplifying an NS3 gene segment of the atypical porcine pestivirus. The invention further provides the kit for detecting the atypical porcine pestivirus and a method for detecting the atypical porcine pestivirus; the kit has the beneficial effects of rapidness and high efficiency, convenience for operation, high specificity, high sensitivity, simplicity for identification, low cost and the like. The primer, the kit and the method, provided by the invention, are used for carrying out quantitative detection and determination on the atypical porcine pestivirus and can be used for rapidly and accurately detecting the copy number of targeting genes and a technological foundation is provided for detecting and identifying the APPV.

Provided in the present invention are a molecule for biomarking cancers and a test composition for screening cancers and a detection method thereof comprising designing a number of oligonucleotide primers or probes by analyzing specimens for methylated regions of targeted genes PAX1, ZNF582, SOX1 and NKX6-1 in physical examination, and then using the oligonucleotide probes to detect whether methylation exists in the target genes, and further judge the likelihood of the occurrence of cancer. The detection methods for methylation status include methylation-specific PCR, (MSP), quantitative methylation-specific PCR (QMSP), bisulfate sequencing (BS), microarrays, mass spectrometer, denaturing high-performance liquid chromatography (DHPLC), pyrosequencing, and enzyme-linked immunosorbent assay (ELISA).

A bioinformatics process which provides an improved means to detect a MAPK-AP-1 cellular signaling pathway in a subject, such as a human, based on the expression levels of at least three unique target genes of the MAPK-AP-1 cellular signaling pathway measured in a sample. The invention includes an apparatus comprising a digital processor configured to perform such a method, a non-transitory storage medium storing instructions that are executable by a digital processing device to perform such a method, and a computer program comprising program code means for causing a digital processing device to perform such a method. Kits are also provided for measuring expression levels of unique sets of MAPK-AP-1 cellular signaling pathway target genes.

Described herein are RNA-based antisense therapeutics to target antibiotic resistant bacteria. The antisense therapeutics disclosed herein can be useful in re-sensitizing drug-resistant bacteria to antibiotics, as well as developing antibiotics that have bactericidal or bacteriostatic effects on the drug-resistant bacteria or are capable of preventing emergence of antibiotic resistance. Also described herein are methods for re-sensitizing a subject to one or more antibiotics in need thereof, methods for identifying target genes involved in adaptive antibiotic resistance in bacteria, and methods for developing antibacterial antisense therapeutics.

The invention provides a phagemid vector construction method for constructing a phage antibody library and an antibody screening method. Phagemids are filamentous phagemids containing restriction enzyme cutting sites of bovine thrombin between coat protein PIII coding genes and target gene insertion sites. The restriction enzyme cutting sites of bovine thrombin are introduced into the phagemids, and hydrolytic cleavage of protein is carried out, so that the background in which the phagemids are non-specifically eluted is reduced. By using the phagemids of the present invention, the antibody screening efficiency is higher, and the problem of low screening efficiency in a phage display technology in the prior art is solved.

The invention discloses a method and a device for detecting a target gene or an SNP (Single Nucleotide Polymorphism) site of the target gene by combining asymmetric PCR (Polymerase Chain Reaction) with a magnetic sensitive detector. The method for detecting the gene marker by combining the asymmetric PCR with the magnetic sensitive detector comprises the following steps: (1) providing a biological sample and treating the biological sample; (2) carrying out asymmetric PCR (Polymerase Chain Reaction) amplification to obtain a single-chain gene marker with a biotin label; (3) hybridizing a single-chain gene marker, and combining the single-chain gene marker with the modified magnetic beads; and (4) detecting a signal through a magnetic sensitive detector. According to the method provided by the invention, the target gene can be directly amplified from the blood, special DNA extraction and purification are not needed, and the reaction time for obtaining the target gene from the blood can be shortened. The asymmetric PCR system provided by the invention can simultaneously amplify and detect a plurality of target genes, the target genes are single-stranded DNA with markers, the influence of complementary chains on the hybridization reaction of the capture probe is reduced, the single-stranded DNA is easy to hybridize and combine with the capture probe, and the intensity of an output signal is improved.

The present invention relates to a miRNA related to oxidative stress damage of duck intestinal mucosa and an application thereof. The miRNA is included, the miRNA is the miRNA (miRNA-218-5p) down-regulated during a stress process, a nucleotide sequence of the miRNA is shown as SEQ ID No.1, a target gene of the miRNA is aldo-keto reductase 7 (AKR7A2), the miRNA is down-regulated-expressed during anoxidative stress process, and by up-regulating expression of the aldo-keto reductase 7 (AKR7A2) gene, oxidative stress response of intestinal epithelial cells is participated. The provided miRNA (miRNA-218-5p) has a very good correlation with the oxidative stress damage, and by verification of effectiveness and molecular biology, expression quantity of the miRNA-218-5p in samples in a stress group and a control group is extremely significant, thus early assisted selection is realized.

The invention discloses a corn genetic transformation method. The corn genetic transformation method comprises the following steps: S1, selecting seeds, cleaning and disinfecting; s2, placing the seeds in a culture medium to germinate the seeds; s3, performing multiplication culture on agrobacterium tumefaciens to obtain a bacterial solution, culturing for 4-6 hours under the conditions that the temperature is 26 DEG C and the oscillation rotating speed is 180-220 rpm, adding a YEB culture solution and acetosyringone until the final concentration is 220 mu M, and culturing for 10-12 hours to obtain an agrobacterium bacterial solution; s4, when the germination length of the seeds cultured in the step S2 reaches 1-3 cm, the seeds are taken out and air-dried, then agrobacterium liquid is injected into the seeds, then the seeds are air-dried, planted into nutrient soil and watered with a nutrient solution, then the seeds continue to be cultured at the temperature of 25 DEG C, and after one week, the seeds are transplanted into a flowerpot to be cultured at the room temperature. Meanwhile, the target gene is introduced into the corn seeds by a method of directly injecting the agrobacterium carrying the target gene into the seeds, so that the experimental period can be greatly shortened, and the transplanting survival rate after transformation is improved.

The invention discloses a soybean powdery mildew resistance gene GmRmd1 encoding protein and application thereof, and belongs to the technical field of gene engineering. The invention provides an application of any one of the following substances in improving powdery mildew resistance of soybeans: 1) a GmRmd1 gene DNA molecule; 2) a GmRmd1 encoding protein; 3) a recombinant plasmid, an expression cassette, a transgenic cell or a recombinant bacterium containing a GmRmd1 gene DNA molecule; the amino acid sequence of the GmRmd1 gene is shown as a sequence 1 in a sequence table. The GmRmd1 gene disclosed by the invention is expressed in soybean varieties susceptible to soybean powdery mildew, so that transgenic soybeans can be endowed with a powdery mildew resistant function; the resistance of powdery mildew can be lost by mutating the GmRmd1 gene in the soybean powdery mildew resistant variety. The gene disclosed by the invention can be introduced into a soybean powdery mildew susceptible variety as a target gene to endow the soybean powdery mildew susceptible variety with powdery mildew resistance, and is of great significance to cultivation of a soybean powdery mildew resistant soybean variety.

The invention discloses a seed capsule specific expression promoter and application thereof and belongs to the technical field of plant genetic engineering. The promoter has a nucleotide sequence as shown by SEQ ID NO.1, or has a complementary nucleotide sequence of the sequence as shown by SEQ ID NO.1; the seed capsule specific expression promoter is a new promoter gene, and is capable of starting the specific expression of a target gene on the seed capsule of plants, therefore the spatial and temporal-specific expression of a foreign gene in the plants is realized; as the promoter is only capable of driving the expression of a downstream target gene in the seed capsule of plants, therefore the adverse effect brought by the prolong expression of the target gene in other tissues of the plants is avoided; the promoter can also be applied to transgenetic seedling screening, is capable of realizing screening in a plant seed stage, and has the advantages of powerful stage and high screening sensitivity.

The invention provides a method for building stable overexpression MGST1 gene cells and primarily studying the biological characteristics of the cells. The method comprises the following steps of (1)synthesizing human MGST1 target gene segments through PCR; (2) connecting the MGST1 target gene segments with eukaryotic plasmid pcDNA3 to form eukaryotic recombinant plasmids; converting the eukaryotic recombinant plasmids into sensory bacterial cells to be cultured; and then selecting monoclone for performing PCR amplification gel electrophoresis to screen out positive clone; (3) extracting thepositive clone screened out from the step (2); analyzing the built eukaryotic recombinant plasmids by using a restriction enzyme double digestion atlas; and performing DNA sequence analysis; and (4) transfecting SPC-A-1 human lung adenocarcinoma cells by the recombinant plasmids pcDNA3-MGST1 with the correct sequence obtained in the step (3); screening SPC-A-1 cells for stably overexpressing MGST1by using G418; and observing the biological characteristics of the cells for stably overexpressing MGST1. The result shows that through the stable overexpression of the MGST1, the cell proliferationcan be promoted; the cell apoptosis can be inhibited; and the cell clone formation capability can be enhanced.