41 results about "Genetic screening method" patented technology

The invention discloses a reference gene screening method for stable expression of suaeda salsa under salt stress. According to transcriptome analysis results obtained in a laboratory and reported reference gene sequence information of other plants, fourteen reference gene candidates with relatively stable expression are preliminarily screened and named as ACT7, ACT1, CCD1, TUA5, UPL1, UBC28, EF1alpha, PP2A, DREB1D, TIM, V-H(+)-ATPase, MPK6, PHT4:5 and CESA1 respectively. For these gene candidates, corresponding amplification primers are designed and reference gene screening is conducted. Tengenes stably expressed in a gene expression profile of suaeda salsa are screened out as the reference genes of suaeda salsa under salt stress, and the stability and reliability of suaeda salsa gene expression analysis and research under salt stress conditions are easily improved. The method makes up for the defect of an existing method in which the use of a single reference gene can lead to errorsand deviations in experimental results, a more reliable conclusion can be obtained, and the method is of great significance for promoting the improvement and perfecting of molecular biological methods and revealing cell differentiation, development and morphogenesis.

The invention provides a genetic screening method for identifying a transfected cell expressing the polypeptide of interest. The methods allows for high throughput screening of recombinant cells for elevated levels of expression of the polypeptide of interest using methylcellulose comprising fluorescent protein A or G to improve detection and cloning. The invention also provides capture media, formulations and methods of making and using thereof.

The invention discloses a screening method of reference genes in a skin tissue of sheep, which relates to the technical field of biological engineering. The screening method comprises the steps that: a method of the reference genes is confirmed; a given amount of RNA (ribonucleic acid) extract is taken and diluted by 2-10 times by ddH2O (double-distilled water); a 50 nanograms / microlitre RNA sample is taken to serve as a template, and a cDNA (complementary deoxyribonucleic acid) synthetic reagent kit is adopted for conducting inverse transcription to synthesize cDNA; the cDNA synthesized through the inverse transcription serves as a template, and six reference genes serve as primers; and finally, pairing variation analysis of normalized factors Vn / n+1 of the six reference genes are calculated based on a geNorm program, so as to decide the amount of the optimum reference genes. The screening method solves the problem that no screening method for GAPDH (reduced glyceraldehyde-phosphate dehydrogenase) genes in the skin tissue of a Chinese merino (Xinjiang type) serving as the reference genes is not available at present. Through the combination of the specificity of the SYBR Green I with double chain DNA to generate fluorescence, a real-time fluorescent quantitative PCR (polymerase chain reaction) detection method of the SYBR Green I for the GAPDH genes in the skin tissue of the Chinese merino (Sinkiang type) is established.

A system in which a ligand is formed by the expression of a polypeptide that converts a ligand precursor into a ligand, and the ligand thus formed binds to a nuclear receptor to thereby induce the expression of a reporter gene located downstream of the target sequence is constructed. Searching a gene library using this system can isolate a gene encoding a polypeptide capable of converting a ligand precursor into a ligand. This system, which takes the advantage of the transcriptional regulatory function of a nuclear receptor, enables screening a ligand that binds to a nuclear receptor and to examine whether or not a test compound is a ligand that binds to the nuclear receptor, and also screening genes that encode polypeptides capable of converting an inactive form of a wide range of transcriptional regulatory factors into an active form.

The invention provides a screening method for a negative regulatory factor of a novel biosynthesis gene cluster of streptomyces. The screening method comprises the following steps: constructing a promoter-mediated reporting system of genes of self-interest in a streptomyces cell, then using a random mutation system based on a transposon Himar1 to mutate the streptomyces with the reporting system,intensively screening a mutant streptomyces strain to obtain a streptomyces strain with high expression of target genes, performing bacteriophage packaging on genomes of the streptomyces strain with high expression of target genes and screening out cosmids with randomly inserted fragments, sequencing DNA of the cosmids to determine the location of the randomly inserted fragments in the genomes ofthe streptomyces strain with high expression of target genes. The screening method, provided by the invention, is stable in screening environment, high in screening flux, low in false positive, high in efficiency, accurate and convenient to operate, and can be widely used in the field of high-yield screening and transformation of industrial streptomyces metabolites, and the screened and transformed streptomyces metabolites are high in yield, good in genetic stability and suitable for industrial production.

The invention relates to a kit for screening dilated cardiomyopathy. The detection genes of the kit are MYBPC3, SCN5A, MYH7 and MYPN. The present invention also provides the application of four genes MYBPC3, SCN5A, MYH7 and MYPN in preparing a kit for screening dilated cardiomyopathy. The advantages of the present invention are: the present invention screens out the most susceptible 4 sporadic dilated cardiomyopathy pathogenic genes in Chinese Han population, and develops an effective early diagnosis method and diagnostic kit for sporadic dilated cardiomyopathy in Chinese Han population, Effectively improve the early detection rate of patients with dilated cardiomyopathy, so as to achieve early treatment and improve the survival rate of patients; compared with the current expensive and inefficient genetic screening methods for dilated cardiomyopathy, the discovery of these four high-risk disease-causing genes It will make screening more targeted, thereby reducing testing costs and improving detection efficiency.

The invention discloses a method of screening reference genes to fluorescently quantitatively detect oviduct of pregnant cows and application of the method and relates to the technical field of animalmolecular biology. Suitable reference genes for the study on genetic expression of oviduct tissue of pregnant cows; the accuracy of fluorescent quantitative detection of the genetic expression of oviduct of the is forcibly guaranteed; important basis is provided for the further study on the regulatory mechanism of the oviduct of cows in early embryonic development phase.

An infectious etiologic agent detection probe set which detects an infectious etiologic agent gene, includes a plurality of kinds of probes including oligonucleotide having base sequences selected from each of a plurality of groups selected from a first group including base sequences of SEQ ID Nos. 1 to 14 and complementary sequences thereof, a second group including base sequences of SEQ ID Nos. 15 to 24 and complementary sequences thereof, a third group including base sequences of SEQ ID Nos. 25 to 36 and complementary sequences thereof, a fourth group including base sequences of SEQ ID Nos. 37 to 47 and complementary sequences thereof, a fifth group including base sequences of SEQ ID Nos. 48 to 57 and complementary sequences thereof, a sixth group including base sequences of SEQ ID Nos. 58 to 68 and complementary sequences thereof, a seventh group including base sequences of SEQ ID Nos. 69 to 77 and complementary sequences thereof, an eighth group including base sequences of SEQ ID Nos. 78 to 85 and complementary sequences thereof, a ninth group including base sequences of SEQ ID Nos. 86 to 97 and complementary sequences thereof, and a 10th group including base sequences of SEQ ID Nos. 98 to 106 and complementary sequences thereof.

The invention discloses a high-throughput functional gene screening method and system for quantitative analysis of cell phenotype images, and belongs to the technical field of gene screening. The images of cells with phenotypes to be screened and those without phenotypes to be screened are captured by a fully automatic fluorescence microscope; converted into black and white binary images of phenotypes to be screened and without phenotypes to be screened respectively; and then divided into individual phenotypes to be screened The image of the cell and the image containing a single cell without a phenotype to be screened; the corresponding part of the image containing a single cell with a phenotype to be screened in the image of a cell with a phenotype to be screened is used as a positive training set, which will contain a single cell without a phenotype to be screened The corresponding part of the cell image in the cell image without the phenotype to be screened is used as a negative training set, and the final model that can identify the cell phenotype is obtained; the image of the gene knockout or overexpression cell to be screened is identified by the final model, and the sample to be screened is obtained. Screening for association of genes with phenotypes. The method improves screening efficiency and accuracy.

Canavan disease, an autosomal recessive leukodystrophy, is caused by deficiency of aspartoacylase and accumulation of N-acetylaspartic acid in brain. Human aspartoacylase (ASP) cDNA spanning 1,435 bp has been cloned and expressed in E. coli. A base change, a854>c, has been found in 85% of the 34 Canavan alleles tested so far, which results in a missense glu285>ala mutation that is predicted to be part of the catalytic domain of aspartoacylase. The invention therefore provides nucleic acid sequences, genes, polypeptides, antibodies, vectors containing the gene, host cells transformed with vectors containing the gene, animal models for the disease, methods for expressing the polypeptide, genetic screening methods and kits, diagnostic methods and kits, methods of treating Canavan disease and methods of genetic therapy for the disease.

The invention discloses a gene rapid screening method capable of simultaneously detecting contaminating bacteria in fruit juice drinks. The method includes the cultivation of juice beverage contaminating bacteria and DNA template extraction, and then through the combination of high-throughput multiplex PCR technology and high-resolution capillary electrophoresis separation technology, it is possible to detect whether the sample contains of contaminating bacteria. Compared with other current molecular biology detection methods based on PCR technology, the present invention can integrate and detect 4 kinds of fruit juice drink contaminating bacteria in one reaction, saving detection cost and detection time, and the detection time is reduced from 7 to 14 days shortened to 8h.

The invention discloses a rapid gene screening method for simultaneously identifying multiple heat-resistant bacteria in fruit juice drinks. The method uses a primer system consisting of 4 pairs of primers, and combines high-throughput multiplex PCR technology with high-resolution capillary electrophoresis separation technology to detect the presence of heat-resistant bacteria in fruit juice beverages. . Compared with other current molecular biology detection methods based on PCR technology, the present invention can integrate and detect 4 kinds of fruit juice beverage heat-resistant bacteria, cycloaliphatic bacillus, cladosporium, spoilage mold, and silk mold in one reaction. Flowers, saving the detection cost and detection time, the detection time was shortened from 7 to 14 days to 8 hours.