53 results about "Transcript level" patented technology

The present invention is directed to nitrogen responsive promoter sequences and promoter control elements, polynucleotide constructs comprising the nitrogen responsive promoters and control elements and methods of identifying the nitrogen responsive promoters, control elements, or fragments thereof. The invention further relates to the use of the present nitrogen responsive promoters or promoter control elements to modulate transcript levels.

The present invention is directed to root active promoter sequences, polynucleotide constructs comprising the root active promoters and methods of identifying the root active promoters, or fragments thereof. The invention further relates to the use of the present root active promoters to modulate transcript levels.

The present invention cloned a cDNA clone encoding isopentenyl diphosphate (hereafter "IPP") isomerase (EC 5.3.3.2) from a cDNA library of Hevea brasiliensis latex. The clone has a continuous open reading frame encoding a peptide of 234 amino acids with a predicted molecular mass of 26.7 kDa. The deduced protein is acidic with an isoelectric point of 4.7 and shows high sequence identity with other IPP isomerases. The recombinant protein expressed in Escherichia coli showed IPP isomerase activity. In vitro rubber biosynthesis assays using washed rubber particle (WRP) deprived of initiating allylic diphosphates were performed with the addition of IPP isomerase in the reaction mixture. Results revealed that the recombinant IPP isomerase is catalytically active in catalyzing the conversion of IPP to DMAPP, a key activation step of the basic five-carbon isoprene unit in rubber biosynthesis. Southern analysis indicated that the IPP isomerase is encoded by two genes in Hevea rubber tree. In Northern blot analysis, two different sizes of transcripts (1.2 and 0.6 kb) were detected from leaf tissues while only one hybridizing band (1.0 kb) was detected from latex. Analyses of RNA extracted from extruded latex and leaf tissues of the trees wounded with nails showed that wounding did not change the transcript level of IPP isomerase.

According to the invention, delta-12 desaturase gene (FADS12, d12) from mortierella alpine ATCC32222 (M. alpina ATCC32222) is amplified, such that multi-copy expression plasmid with inserted opai / d12 co-expression gene and recombinant Yarrowia lipolytica yeast strain are prepared. According to the invention, the recombinant Yarrowia lipolytica yeast strain assists in increasing endogenous substrate linoleic acid (LA) amount and outer soybean oil increased substrate LA amount in fermentation medium, through expressing d12. In the strain, opai gene and d12 gene expression and transcript levels are increased with a multi-copy integration manner, such that conjugated linoleic acid t10 and c12-CLA specific isomer synthesis amount in recombinant Yarrowia lipolytica yeast strain can be further improved.

Described is a method for determining a nucleotide sequence within cDNA, the frequency of a nucleotide sequence in a cDNA sample, as well as a method for (unbiased) determination of relative transcript levels of genes without sequence information of these genes being required, said methods using complexity reduction and (high throughput) sequencing.

The invention belongs to the biotechnology field, relates to application of PSMD4 protein, and in particular relates to application of the PSMD4 protein in preparation of a liver cancer prognostic evaluation kit. The invention adopts an immunohistochemistry technology, a microscope photograph technology and computer image processing software to test the relative transcript level of PSMD4 protein of tumor tissues relative to the tumors and determine the correlation between the relative transcript level of PSMD4 protein and the prognosis of hepatocellular carcinoma patients after operation by a manner of combining the postoperative follow-up information. The PSMD4 protein can be used for preparing a protein molecular marker for judging prognosis of the liver cancer patients, and has important instruction significance to postoperative monitor and sequential therapy of the hepatocellular carcinoma patients.

A novel suspension hybridization assay was used to determine nucleic acid copy number by flow cytometry. The assay was validated with low copy (lc) products ranging in length from 100 to 2304 bp conjugated to spectrally-distinct polystyrene microspheres. In the example provided herein, these conjugated microspheres were used as multiplex hybridization probes to detect homologous sequences in genomic DNA extracted from cytogenetic cell pellets and labeled with biotin-dUTP. Hybridization was detected with phycoerythrin-labeled streptavidin and analyzed by flow cytometry. Copy number differences were distinguishable by comparing the mean fluorescence intensities of test probes with a diploid reference probe in genomic DNA of patient samples and abnormal cell lines. The assay is capable of distinguishing a single allele and three alleles at a test locus from a biallelic reference sequence, regardless of chromosomal context. The assay is an improvement on previous methods which require prior amplification of locus-specific target DNA because, lc probes provide adequate specificity and sensitivity for accurate copy number determination of homologous targets. Because of its high sensitivity and accuracy, the assay is useful for determination of nucleic acid copy number for a variety of applications, including determination of genomic copy number in humans, animal models of disease and in solution, measurement of transcript levels, forensic DNA analysis, and quality control analysis in agriculture.

The invention relates to an extraction method of protein from animal tissues, in particular to an extraction method of total protein from a rumen epithelial tissue of a milk cow. The method comprises the steps of putting a rumen epithelial tissue sample of the milk cow in a precooled mortar, adding liquid nitrogen, grinding into powder, adding an acetone solution of precooled trichloroacetic acid, oscillating, eddying, uniformly mixing for overnight aging, centrifugally collecting sediment, freeze-drying the tissue sediment, adding lysate, incubating and eddying at intervals, and centrifugally collecting supernate, namely the total protein of the rumen epithelial tissue of the milk cow. For the total protein of the rumen epithelial tissue of the milk cow, extracted by the method, a total protein mixture can be separated by a two-dimensional gel electrophoresis technology according to an isoelectric point and molecular weight of the protein, the relative abundance of the protein is acquired, the isoelectric point, the molecular weight and a relative transcript level of each protein can be clearly and visually shown in a gel atlas, and the method has the advantages of more obvious integrity and visualization in comparison with the prior arts such as a high efficiency liquid chromatography separation method, an enzyme-linked immunosorbent assay and immunoblotting.

The invention discloses a method for monitoring phenanthrene pollution in water through a gene P4501A (CYP1A). Fishes serve as biological samples for monitoring water pollution, and the gene CYP1A of the fishes serves as a sensitive biomarker for monitoring the phenanthrene pollution in water. The method has the advantages that the gene CYP1A serves as the sensitive biomarker for monitoring water pollution, the degree of water pollution is monitored by testing the relative transcript level of the gene CYP1A, and the method can sensitively reflect whether water is polluted or not and the degree of water pollution and even can sensitively detect the water pollution of a micro-pollution source; the perniciousness and the damage degree of phenanthrene pollution can be sensitively monitored from the biological angle, and an accurate, quick and scientific method is provided for water ecological environment monitoring, toxicology diagnosing, regulating, preventing and treating.

The present invention provides a TOP2A inhibition by temozolomide useful for predicting glioblastoma patient's survival. Glioblastoma (GBM) is the most common, malignant primary adult brain tumor. The conventional treatments for GBM, include surgery, radiation, and chemotherapy which have only modestly improved patient survival. The patients with GBM expressing higher TOP2A transcript levels had better prognosis. More interestingly, the present invention reports that temozolomide is an inhibitor of TOP2A activity in vitro. The present invention further shows that siRNA knock down of TOP2A rendered a glioma cell line resistant to temozolomide chemotherapy.; Thus it is demonstrated for the first time that temozolomide is a TOP2A inhibitor and establishes that TOP2A transcript levels determines the chemosensitivity of glioblastoma to temozolomide therapy thus explaining the very high levels of TOP2A transcript being a good prognostic indicator in GBM patients receiving temozolomide chemotherapy.

The invention belongs to the technical field of cancer molecular diagnosis technique, in particular to a marker for identifying benign and malignant pulmonary nodules by using an exosome protein and an application of the marker. The marker comprises the following steps: S1, looking for differentially expressed proteins of exosomes in plasmas of patients suffering from benign and malignant pulmonary nodules; S2, screening the differential proteins; S3, verifying the differential diagnosis effect of FGB on the benign and malignant pulmonary nodules; and S4, analyzing the relative transcript level. The non-invasive liquid-state biopsy method is used to search out the molecular marker for identifying the benign and malignant pulmonary nodules, and the mass spectrometric detection and the subsequent bio-information analysis show that the molecular marker FGB provides a novel molecular target for the diagnosis of the pulmonary nodules and the observation of clinical effects.

The present invention is directed to promoter sequences and promoter control elements, polynucleotide constructs comprising the promoters and control elements, and methods of identifying the promoters, control elements, or fragments thereof. The invention further relates to the use of the present promoters or promoter control elements to modulate transcript levels.

The present invention is directed to drought responsive promoter sequences, polynucleotide constructs comprising the drought responsive promoters and methods of identifying the drought responsive promoters or fragments thereof. The invention further relates to the use of the present drought responsive promoters to modulate transcript levels.

The invention discloses a method for rapidly evaluating rice SRBSDV variety resistance through detecting a viral multiplication speed in coleoptile. The method comprises the following steps: (1) selecting an identifying scaleplate; (2) performing hypoxia induction on the rice to be tested and identifying the coleoptile of the scaleplate variety; (3) inoculating the toxic sogatella furcifera on the coleoptile and performing dark culture; (4) performing fluorogenic quantitative PCR to obtain a relative transcript level (RQ value) of an SRBSDV virus S9 and a reference gene in each rice variety period time; (5) drawing an SRBSDV proliferation index trend line; and (6) evaluating the resistance level and the resistance grade of the rice variety to be tested on the Southern rice black-streaked dwarf virus according to the SRBSDV proliferation index trend line formula. By detecting the viral multiplication speed in the coleoptiles and rapidly evaluating the resistance of the rice SRBSDV germplasm resistance, the resistance level and the resistance grade of the rice variety on the SRBSDV from the molecular level in the rice germination stage are determined, the identification period is short and the reliability is high, and the method has big breeding application value.

Disclosed are two methods of treating or preventing a proliferative disease as characterized and / or diagnosed by at least one selected from an accumulation of branched-chain amino acid(s) (BCAA), suppression of activity or transcripts level of BCAA catabolic enzyme(s) and a decrease in acylcarnitine level (C5:1), one comprising the administration of a BCAA catabolism enhancer and / or branched chain alpha-ketoacid dehydrogenase complex (BCKDC) kinase inhibitor and the other comprising administration of a meal replacement with low BCAA levels. In three separate embodiments, methods of diagnosis or prognosis are disclosed, each comprising the measurement of the levels of a different marker, i.e. BCAA, BCAA catabolic enzymes or acylcarnitine (C5:1). In preferred embodiments, the proliferative disease is cancer. Also disclosed is a kit or microarray chip useful for methods thereof.

The invention discloses a method for screening a fine progeny strain polyploidy plant of Chinese gooseberry Hort 16A, and relates to the screening of a fine strain polyploidy of the plant. The method comprises the following steps: extracting total RNA of a leaf of a tissue culture seedling, reversely transcribing RNA into cDNA, selecting 1 to 2 differential genes having an up-regulation trend in transcriptomes of diploid and polyploidy plants, taking a housekeeping gene Elongation factor EC as a reference gene, fluorescently quantifying a PCR (Polymerase Chain Reaction), drawing an amplification curve and a solubility curve for the differential genes, selecting the differential genes, calculating a relative transcript level, and taking the tissue culture seedling, whose differential genes are more than twice those of the diploid, as the tissue culture seedling of the polyploidy. The theory and the method for screening the polyploidy variant plant are innovated, and the efficient feasible method is provided for screening the polyploidy plant of the Chinese gooseberry.

The invention provides oligonucleotide and a method for joint detection of relative transcript levels of genes PTEN (phosphatase and tensin homolog deleted on chromosome ten) and VEGF (vascular endothelial cell growth factor) based on fluorogenic quantitative PCR (Polymerase Chain Reaction), mainly relating to a forward primer PTEN-F, a reverse primer PTEN-R and a probe PTEN-Probe for detecting the gene PTEN; an forward primer VEGF-F, a reverse primer VEGF-R and a probe VEGF-Probe for detecting the gene VEGF; and a forward primer Actin-F, a reverse primer Actin-R and a probe Actin-Probe for detecting a reference gene actin. The relative transcript levels of the detected genes PTEN and VEGF are respectively compared with reference values of the relative transcript levels of the genes PTEN and VEGF of a healthy person, and the comparison result can be used for auxiliary diagnosis of various kinds of cancers.

The present invention provides a method for producing transgenic Cotton plants. In one method transformed plants, that overexpress the transgene shows a phenotype that includes increased boll number, size and lint percentage in compare to the wild type plants; whereas in the second method transformed plants that reduced the transgene level produced plants with decreased number of cotton boll, size and lint percentage in compare to wild type and overexpression line both. q-RT PCR analysis showed that transgene transcript level was higher at fiber initiation stage (0 DPA) after that its level decreases throughout all developmental stages.

Although metabolic networks have been reconstructed on a genome-scale, the corresponding reconstruction and integration of governing transcriptional regulatory networks has not been fully achieved. Here such an integrated network was constructed for amino acid metabolism in Escherichia coli. Analysis of ChlP-chip and gene expression data for the transcription factors ArgR, Lrp, and TrpR showed that 19 / 20 amino acid biosynthetic pathways are either directly or indirectly controlled by these regulators. Classifying the regulated genes into three functional categories of transport, biosynthesis, and metabolism leads to elucidation of regulatory motifs constituting the integrated network's basic building blocks. The regulatory logic of these motifs was determined based on the relationships between transcription factor binding and changes in transcript levels in response to exogenous amino acids. Remarkably, the resulting logic shows how amino acids are differentiated as signaling and nutrient molecules. This reveals the overarching regulatory principles of the amino acid stimulon.