94 results about "Gene copy" patented technology

The invention belongs to the technical field of lake and reservoir water source risk prediction, and provides a lake and reservoir water source algae bloom risk early warning system based on a 16S / 18SrRNA gene expression quantity threshold, and the method comprises the following steps: S1, collecting a sample from a lake and reservoir water source surface layer water body; s2, using 7-gate water bloom algae specific primers for blue-green algae, green algae, diatom, euglena, dinoflagellate, chrysophyta and cryptoalga; s3, based on the qPCR standard curve, calculating the copy number of the 16SrRNA / 18SrRNA gene of the seven water bloom algae; s4, when the gene expression quantity of a certain algal bloom algae continuously reaches 106-7 copy number / mL for 7-12 days, determining that the algal bloom algae is in a window phase; according to the threshold value of the gene expression quantity of the water bloom algae 16SrRNA / 18SrRNA, the window period of water bloom algae cells can be accurately recognized, so that early warning of the algae bloom risk in the lake and reservoir water source is achieved, and early warning of algae bloom outbreak in the lake and reservoir water source is achieved one week or above in advance.

The invention provides a primer probe combination and a kit for detecting the copy number of a 22q11 region based on digital PCR and a use method of the primer probe combination. Specific probes of a target gene TBX1, a target gene CRKL, a target gene SMARCB1 and a reference gene GAPDH are integrated in different fluorescence channels, and the copy number of the target gene is detected according to the ratio of the copy number of the reference gene to the copy number of the three target genes. According to the invention, amplification products of different sites in different fluorescence channels are integrated in the same system in digital PCR reaction, so that the detection flux is greatly improved, and low-cost and high-flexibility accurate quantification of 22q11. 2 copy number variation is realized through a single-channel multi-target fluorescence layering strategy; and the dependence on expensive equipment, complex operation and professional analysis in the prior art is overcome.

According to the application of the HTATSF1 gene in regulation and control of cattle muscle growth and development and breeding, the influence of the HTATSF1 gene on muscle growth and development and the correlation between HTATSF1-CNV and cattle growth traits are verified, a reliable molecular marker is provided for cattle seed selection and breeding, and help is provided for cattle variety improvement. A foundation is laid for association between copy number variation and growth traits of the cattle HTATSF1 gene, cattle molecular marker-assisted selection breeding work can be accelerated, the method is simple, rapid and convenient to popularize and apply, and a complete solution from fundamental research to industrial application is provided for improvement of cattle germplasm resources.

The invention discloses a probe group for detecting copy number variation of pathogenic genes of limb development defect diseases as well as a design method, a kit and application of the probe group, and belongs to the technical field of biomedicine. The probe group comprises probes of which the nucleotide sequences are as shown in SEQ ID NO. 1 to SEQ ID NO. 516. The invention also discloses a design method of the probe group, a kit comprising the probe group, and a method for detecting copy number variation of the pathogenic gene of the limb development defect disease by using the probe group or the kit. The probe group and the kit provided by the invention can realize high-throughput, high-resolution, high-accuracy and low-cost copy number variation detection.

The application provides a standard plasmid for detecting the copy number of exogenous genes of a recombinant CVA10 vaccine, and a preparation method and application thereof. The standard plasmid of the application comprises an endogenous gene MOX fragment SEQ ID No. 1, an exogenous gene P1 fragment SEQ ID No. 2 and an exogenous gene 3CD fragment SEQ ID No. 3 which are connected in series in a cloning vector. The application constructs the standard plasmid, adopts Taqman probe-fluorescence quantitative PCR method, and simply, quickly, absolutely quantitatively and accurately analyzes the copy number of inserted exogenous genes, so that the detection time of the determination of the copy number of exogenous genes of the recombinant CVA10 vaccine is effectively shortened, repeated operation is facilitated, and the determination of the copy number of exogenous genes of the recombinant CVA10 vaccine based on the Hansenula platform and the verification of the genetic stability of the exogenous genes of the recombinant CVA10 vaccine have a wide application prospect.

A method and device for predicting a genotype of a target gene, the method comprising: a baseline correction step; a feature extraction step, comprising obtaining a cluster center of a specific region of the target gene according to a corrected depth of each capture site, and taking the cluster center as a feature value of the corresponding region; and a prediction step, comprising predicting a genotype of the target gene in sequencing data of a sample to be tested according to the feature value. The method has the advantages of fast analysis speed, high accuracy, wide application range, no need for reference set correction, avoidance of the influence of sequencing fluctuation on the analysis result, ease of implementation of high-throughput screening typing at a population level, and applicability to individualized medication guidance for more populations and acceleration of large-scale research and development of pharmacogenomics.

The present invention provides a method for efficiently delivering a transgene to the brain of a subject. The method includes intrathecal administration or one or more injections into the striatum of a replication defective herpes simplex virus (rdHSV) vector comprising an inactivating mutation of one copy of infected cell protein 0 (ICP0) HSV gene; and wherein the rdHSV-1 vector expresses a functional ICP0 protein from the other copy of the ICP0 gene.

The application provides a primer probe composition for detecting genetic stability of cells and application thereof, the primer probe composition comprises primer pairs SEQ ID NO:1-2 and a probe SEQ ID NO:3 of a VEGF light chain gene, and further comprises primer pairs SEQ ID NO:4-5 and a probe SEQ ID NO:6 of a VEGF heavy chain gene. The application quantitatively detects the antibody light and heavy chain gene copy numbers in cells of different culture generations by fluorescence quantitative PCR based on a Taqman probe method, extracts cell genomic DNA and performs PCR amplification, and prepares a target gene standard product from the amplification product, thereby significantly improving the specificity and accuracy of quantitative detection of the gene copy number.

The invention belongs to the technical field of biology, and provides endoglucanase FsCel45B of a GH45 family, a related biological material and a high-efficiency expression method of the endoglucanase FsCel45B. The GH45 family endoglucanase FsCel45B comprises the following A1) or A2): A1) a protein with an amino acid sequence as shown in SEQ ID NO: 1; a2) a protein which is obtained by substitution and / or deletion and / or addition of amino acid residues on the amino acid sequence shown in SEQ ID NO: 1 and has the same function as the protein shown in A1). The invention further provides an expression optimization strategy in pichia pastoris, based on a pichia pastoris expression vector pPICZ alpha A, an AOX1 promoter and an alpha-MF signal peptide sequence are replaced and co-expressed with disulfide bond folding isomerase (PDI) and a vesicle transport related gene Ssa4, the expression quantity is improved by combining with the increase of the gene copy number, and the expression optimization strategy has industrial production potential.

The invention discloses a TaqMan probe qPCR (quantitative polymerase chain reaction) detection primer group for detecting pathogenic vibrio alginolyticus, a kit and application, and belongs to the technical field of molecular biology and pathogenic microorganism detection. The method comprises the following steps: designing a pair of specific primers and probes by taking the vops gene of the vibrio alginolyticus as a new target, connecting the vops gene with plasmids by amplifying the vops gene, constructing a recombinant plasmid standard substance, carrying out qPCR detection on the copy number of the vops gene of the vibrio alginolyticus by utilizing the designed specific primers and probes, and establishing a corresponding relationship between the vibrio alginolyticus and the vops gene. And the vibrio alginolyticus can be quantified through the copy number of the vops gene. On the basis of novel molecular target detection, no amplification signal exists for a detection sample which does not contain vibrio alginolyticus, and the specificity is good; the method is rapid and accurate in detection and high in sensitivity, the lower detection limit is as low as 100-level copies, non-specific amplification can be avoided, and operation is simple and rapid.

A generic cancer prognosis evaluation system based on LAMA5 expression provided by the invention relates to the technical field of generic cancer prognosis evaluation, and comprises the following steps: acquiring multi-source molecular data, establishing a standardized data input path to obtain LAMA5 gene molecular information, and completing sample identification and field standardization by calculating mutation frequency and gene copy number variation of the LAMA5 gene molecular information to obtain a generic cancer prognosis evaluation result. And an LAMA5 generic cancer molecular database is formed. A standardized data input path and a unified field format are formed by establishing an LAMA5 generic cancer molecular database and integrating TCGA, GTEx, a human protein map and other multi-source molecular data, the database achieves generic cancer prognosis evaluation system calculation of LAMA5 mutation frequency and gene copy number variation, the data consistency and statistical reliability of multiple cancer species are improved, and the method is suitable for large-scale popularization and application. And a data basis is provided for pan-cancer level expression and variation analysis.

The invention relates to the technical field of microbial fermentation and genetic engineering, and particularly discloses a yeast engineering bacterium for efficiently producing plastic depolymerases and application of the yeast engineering bacterium. The engineering yeast takes pichia pastoris as an expression host, a molecular chaperone is co-expressed, a vacuole sorting receptor gene is knocked out or knocked down, in addition, a yeast cell factory for efficiently expressing the plastic depolymerases is constructed by further combining regulation and control element optimization and target gene copy number regulation, and the engineering yeast has the advantages that the engineering yeast can be used for efficiently expressing the plastic depolymerases; the regulatory element comprises any one or a combination of more of a strong promoter, a signal peptide and a C-terminal tag. The engineering yeast provided by the invention can realize efficient accumulation and secretion of the plastic depolymerases, obviously improve the enzyme yield and reduce the production cost of unit enzyme activity, and provides a stable and efficient yeast expression system for industrial and low-cost production of the plastic depolymerases; the method has an important application prospect in the fields of biodegradation and resource utilization of waste plastics.

This invention relates to the field of nucleic acid detection technology, specifically disclosing a liquid-phase nucleic acid chip for isothermal amplification of common algal blooms and its application. The method includes: collecting water samples and filtering to enrich algal cells; lysing the algal cells using cell lysis buffer to extract DNA; adding the extracted DNA to the liquid-phase nucleic acid chip for isothermal amplification; detecting the Ct value using a qPCR instrument; and determining the species and cell density of the algal bloom based on a preset standard curve. This invention directly quantifies algal cell density using algal gene copy number, with significantly higher accuracy than microscopic counting, and does not rely on the experience of professional personnel. It can be widely applied in frontline water resource management work.

The application belongs to the technical field of biological information detection, and discloses a gene copy number variation detection method, according to the confidence interval determination of copy abnormal sites, clustering, determining the copy abnormal sites belonging to the high copy variation sub-class and the RD value being greater than the corresponding confidence interval and the copy abnormal sites belonging to the low copy variation sub-class and the RD value being less than the corresponding confidence interval as the copy variation sites, merging the copy variation sites with adjacent positions and the same variation type to obtain more accurate CNV fragments, separating the interval size of the RD site and the interval size of the CNV fragment, so that the resolution of the RD site can be improved while the stability of the CNV fragment is ensured, and the unsupervised machine learning method is used for the clustering classification of the copy abnormal sites and the comparison with the confidence interval, so that the copy variation sites are double-recognized, the recognition accuracy of the copy variation sites is further improved, and the accuracy, sensitivity and stability of the CNV detection are improved.

The application discloses a method for identifying direct discharge of domestic sewage and application thereof. The method comprises the following steps: detecting the copy number concentration of bacteriophage marker genes and Bacteroides marker genes of adjacent two days of a water body to be detected respectively, calculating the ratio of the copy number concentration of the bacteriophage marker genes and the copy number concentration of the Bacteroides marker genes per day, and calculating the ratio between the two days of the water body to be detected, and combining a specific ratio threshold to determine whether the direct discharge of domestic sewage appears in the water body to be detected, and the day when the direct discharge of domestic sewage appears can be accurately determined, so that the direct discharge of domestic sewage can be traced and controlled; wherein the bacteriophage marker gene is CPQ_064, and the Bacteroides marker gene is BacHum, BacH, HuBac and / or HF183.

Provided is a method for preparing a recombinant human collagen. The method comprises: constructing a genetically engineered bacterium which expresses a prolyl hydroxylase and a recombinant human collagen; culturing the genetically engineered bacterium and performing digestion treatment by means of pepsin to obtain a recombinant human collagen type I or type III. The prolyl hydroxylase comprises P4Ha2, P4Hb and vP4H, and by means of controlling the ratio of the copy numbers of the genes thereof, the recombinant human collagen and a natural human collagen are consistent in hydroxylation modification. Also provided is a large-scale production method for the recombinant human collagen. The prepared recombinant human collagen has completely the same sequence composition and length and substantially the same hydroxyproline content as the natural human collagen, and also has a stable full-length triple helix structure and biological activity without immunogenicity and potential risk of viruses.

The invention discloses application of an HAVCR2 gene in cattle molecular marker-assisted selective breeding, and belongs to the technical field of molecular genetics. The method comprises the following steps: by taking cattle genome DNA as a template, amplifying a cattle HAVCR2 gene copy number variation region by utilizing a specific PCR primer, amplifying a cattle BTF3 gene partial segment as an internal reference, and finally calculating and judging the individual copy number variation type by utilizing a 2 * 2-delta delta Ct method. The method provided by the invention lays a foundation for establishing association between cattle HAVCR2 gene copy number variation and growth and development, can be used for cattle molecular marker-assisted selective breeding work, and is convenient to popularize and apply.

This invention relates to the field of genetic engineering technology and provides a method for increasing erythromycin yield by modifying the SACE_5680 gene of *Rhodotorula sacchariformis*. The method involves deleting the GntR family SACE_5680 gene in *Rhodotorula sacchariformis* through genetic engineering, thereby obtaining a high-yielding engineered strain of *Rhodotorula sacchariformis* for erythromycin production. Erythromycin is then produced by fermentation using this high-yielding engineered strain. The nucleotide sequence of the SACE_5680 gene is shown in SEQ ID NO. 1. This invention also provides an application of the above method. In this study, the negative regulator of erythromycin biosynthesis, SACE_5680, was screened. By deleting a copy of the SACE_5680 gene on the chromosome of *Rhodotorula sacchariformis* through genetic engineering, a high-yielding strain of erythromycin can be obtained, providing technical support for increasing erythromycin fermentation yield in industrial production.

Disclosed is a method of determining whether a human subject is not a carrier of spinal muscular atrophy (SMA). This method includes the steps of (i) collecting a genomic deoxyribonucleic acid (DNA) sample from a human subject; (ii) screening the genomic DNA sample to determine the human subject's copy number of survival of motor neuron 1 (SMN1) gene and whether one of the copies of the SMN1 gene is positive for a polymorphism associated with non-carriers of SMA having two copies of the SMN1 gene; and (iii) determining the human subject as not a carrier of SMA if the human subject includes two copies of the SMN1 gene with one of those copies being positive for the polymorphism. Also disclosed is a method of determining whether an individual has a decreased risk of being a carrier of spinal muscular atrophy (SMA), where the individual is identified to have a decreased risk of being a carrier of SMA when the individual has two copies of the SMN1 gene with one of those copies being positive for the polymorphism.

The application provides a STRC gene copy number variation detection method based on whole genome sequencing. The STRC gene and the STRCP1 gene are subjected to sequence alignment to find each difference site of the STRC gene and the STRCP1 gene. For each difference site, the sequence of the corresponding STRC position and STRCP1 position in the genome is read from a variation detection file. The total copy number of the true gene and the false gene is calculated by taking the reference site in the genome as a reference. The STRC gene copy proportion on each difference site is calculated. The STRC gene copy number on each difference site is calculated according to the total copy number and the STRC gene copy proportion. The STRC gene copy number on each exon is determined according to the STRC gene copy number on each difference site. The method can realize the detection of the STRC copy number, simplifies the detection process, improves the detection throughput and reduces the cost.

The invention provides a sheep-derived component rapid quantitative detection method based on dual real-time fluorescent PCR. According to the method, a single-copy nuclear gene GAPDH is taken as a target spot, a sheep specific primer and a probe are designed in a hypervariable region of the single-copy nuclear gene GAPDH, and an internal reference primer and a probe capable of identifying various common animal-derived components are designed in a conserved region of the single-copy nuclear gene GAPDH. As the target and the internal reference are located in the same gene and coexist in the same cell according to the proportion of 1: 1, quantitative deviation caused by gene copy number difference or expression fluctuation in a traditional two-step method is remarkably reduced. The detection strategy constructed by the invention has double advantages of specific recognition and broad-spectrum reference, is clear in quantitative basis and simple and convenient to operate, is suitable for rapid recognition and quantitative adulteration analysis of sheep-derived components in meat products, and has wide application prospects and supervision values.

The invention provides a method for improving the yield of lincomycin by modifying a streptomyces lincomycin SLCG3615 gene, which comprises the following steps: knocking out a TetR family transcriptional regulation gene SLCG3615 in the streptomyces lincomycin through a genetic engineering way to obtain a lincomycin high-yield engineering strain, and fermenting the obtained strain to produce the lincomycin. Wherein the nucleotide sequence of the SLCG3615 gene is as shown in SEQ ID NO. 1. The invention also provides an application of the SLCG3615 gene knockout modified streptomyces lincomus. The lincomycin biosynthesis negative regulator SLCG3615 is screened in the research, the SLCG3615 gene copy on the chromosome of the streptomyces lincomycin is knocked out through a genetic engineering way, a lincomycin high-yield strain can be obtained, and a technical support is provided for improving the fermentation yield of the lincomycin in industrial production.

The invention relates to the technical field of synthetic biology, in particular to a saccharomyces cerevisiae recombinant strain with high gene copy integration and high geraniol yield as well as a construction method and application of the saccharomyces cerevisiae recombinant strain. According to the method, the copy number of geraniol synthase genes is increased through a saccharomyces cerevisiae gene multi-copy integration technology, efficient and stable expression of the geraniol synthase genes is achieved, supply of geraniol precursors is improved by further combining the modes of blocking a precursor competitive pathway and the like, a saccharomyces cerevisiae recombinant strain capable of efficiently and stably synthesizing geraniol is constructed, and the method is suitable for large-scale production of geraniol. Restriction factors of synthesis of geraniol by saccharomyces cerevisiae are removed, and the recombinant strain has high genetic stability and good environmental protection property. The recombinant strain constructed by the invention can be used for efficiently synthesizing geraniol and derivatives of geraniol, and is applied to essence, fragrance and food industries.

The present invention addresses the problem of developing a novel drug for treating osteosarcoma. The present invention also addresses the problem of developing a therapeutic marker for studying the therapeutic effect of a drug in the treatment of osteosarcoma. The present invention shows that in order to solve the above-mentioned problem, a compound capable of inhibiting cell proliferation of osteosarcoma is found, and thus a novel pharmaceutical composition for treating osteosarcoma can be provided. It is also shown that osteosarcoma to be treated, which is a novel pharmaceutical composition, is an osteosarcoma in which the number of copies of an MCL1 gene in a cell has been increased or an osteosarcoma in which the production of an Mcl-1 protein has been increased, or an osteosarcoma in which the number of copies of a BCL2L1 gene has been increased or an osteosarcoma in which the production of a Bcl-xL protein has been increased. Therefore, by detecting these, it can be used as a therapeutic marker for studying the therapeutic effect of a novel pharmaceutical composition for the treatment of osteosarcoma.

The invention belongs to the technical field of biological medicine, and particularly relates to a method for measuring the copy number of a CAR gene through double digital PCR. The invention provides the method for determining the copy number of the CAR gene in the cell therapy product based on the digital PCR, and the method does not need a standard curve, has universality, is suitable for identifying the copy number of any CAR gene containing the WPRE virus vector, and has no limitation on the CAR structure. The primer probe pair is good in specificity, high in sensitivity, good in reaction system repeatability, good in stability and good in application prospect.

Technology provided herein relates in part to non-invasive classification of one or more mosaic copy number variations (CNVs) for a test sample. Technology provided herein is useful for classifying a mosaic CNV for a sample as part of non-invasive pre-natal (NIPT) testing and oncology testing, for example. In particular, a method is provided for classifying presence or absence of genetic mosaicism for a biological sample, the method includes identifying a genetic copy number variation region in sample nucleic acid from a subject, e.g. a pregnant female, determining a fraction of nucleic acid having the copy number variation in the sample nucleic acid, determining a fraction of a minority nucleic acid, e.g. fetal nucleic acid, in the sample nucleic acid, comparing the two fractions to generate a mosaicism ratio, and classifying a presence or absence of a genetic mosaicism for the copy number variation region according to the mosaicism ratio.

The invention provides a method for monitoring a yeast fermentation process and pre-judging product quality. The method comprises the following steps: collecting yeast samples and recording fermentation days; the method comprises the following steps: extracting total DNA (Deoxyribonucleic Acid) of a sample, and respectively and quantitatively detecting the copy number of 16S rRNA genes of bacteria and the copy number of 18S rRNA genes of fungi; inputting the bacterial 16S rRNA gene copy number, the fungus 18S rRNA gene copy number and the fermentation day number into a random forest classification model, and outputting a normal or abnormal fermentation state; and pre-judging the product quality based on the fermentation state. The method breaks through the limitation of traditional dependence on sensory experience and a culture method, the microbial biomass is reflected in real time through absolute quantification of the microbial gene copy number, the detection time is remarkably shortened, and the accuracy of the random forest classification model is extremely high.

The application provides a use of a compound BMS309403 in preparation of a medicament for preventing and treating acute hepatopancreas necrosis disease of prawns, and belongs to the technical field of prawn disease prevention and treatment. It is unexpectedly found that an inhibitor of a fatty acid binding protein LvFABP of Litopenaeus vannamei, i.e. the compound BMS309403, can be effectively applied in the prevention and treatment of AHPND, which can not only reduce the disease of hepatopancreas and intestinal tract of prawns, but also effectively reduce the Vibrio virulence gene copy number of hepatopancreas of diseased prawns, so that the survival rate of prawns infected with acute hepatopancreas necrosis disease can be significantly improved. In addition, as a small molecule compound, the BMS309403 is more superior to small molecule double-stranded RNA in synthesis, preservation and use, and therefore can have a better prospect in the actual production application of AHPND prevention and treatment.

This invention belongs to the field of biotechnology and provides GH45 family endoglucanase FsCel45B, related biological materials, and efficient expression methods. GH45 family endoglucanase FsCel45B includes the following A1) or A2): A1) a protein with the amino acid sequence shown in SEQ ID NO:1; A2) a protein with the same function as the protein shown in A1) obtained by substituting and / or deleting and / or adding amino acid residues from the amino acid sequence shown in SEQ ID NO:1. This invention also provides an expression optimization strategy in Pichia pastoris, based on the Pichia pastoris expression vector pPICZαA, replacing the AOX1 promoter and α-MF signal peptide sequence, and co-expressing it with disulfide bond folding isomerase (PDI) and the vesicle transport-related gene Ssa4, and combining this with an increase in gene copy number to improve expression levels, possessing industrial production potential.

The invention discloses a primer pair for evaluating large yellow croaker cell senescence based on relative telomere length and a method thereof, and belongs to the field of molecular genetics. The specific method comprises the following steps: 1) taking DNA of a large yellow croaker cell genome; 2) performing qPCR amplification on the DNA obtained in the step S1 by using a first qPCR amplification system containing a telomere universal primer to obtain a telomere copy number Cttel; (3) carrying out qPCR amplification on the obtained DNA by using a second qPCR amplification system containing the forward primer as shown in SEQ ID No.1 and the reverse primer as shown in SEQ ID No.2, so as to obtain the copy number Ctactb2 of the internal reference single copy gene; and 4) calculating the relative telomere length T / S = 2-[delta] Ct, wherein [delta] Ct = Ctel-Ctactb2. The relative telomere length detection actb2 primer provided by the invention can specifically amplify large yellow croaker DNA, and can realize accurate detection by adopting a trace sample.