124 results about "Gene copy" patented technology

The invention relates to a method for carrying out pathogenic microorganism identification and gene variation state joint detection on a pulmonary alveolar lavage fluid sample based on high-throughput sequencing. The method comprises the following steps: extracting nucleic acid from the pulmonary alveolar lavage fluid sample; constructing a metagenome capture library, and constructing a lung cancer polygene library; carrying out final library pooling; performing high-throughput sequencing; and analyzing data. Compared with the prior art, the scheme provided by the invention is based on a technical path of'macro 'captured by metagenome and'target' of targeted high-throughput sequencing; after the constructed final library is subjected to high-throughput sequencing and data analysis, index output (pathogenic microorganism identification, host chromosome copy number variation, gene mutation, gene fusion, gene copy number variation and methylation state) of multiple biomarkers can be met in one-time detection.

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 relates to recombinant pichia pastoris for high expression of porcine pancreatic phospholipase A2 and application of the recombinant pichia pastoris, and belongs to the technical field of genetic engineering. According to the method disclosed by the invention, a vacuole sorting receptor Vps10 is knocked out by utilizing a CRISPR-Cas9 system, a CPY sorting way between a trans-golgi apparatus and a vacuole is interrupted, and ppPLA2 degradation is reduced; then, in order to relieve unfolded protein response in the endoplasmic reticulum, molecular chaperones are introduced into the genome, and synthesis of ppPLA2 is enhanced; the expression intensity of the ppPLA2 is improved by increasing the gene copy number of the ppPLA2; and finally, high-density fermentation is performed in a 3-L bioreactor, so that high-efficiency expression of the ppPLA2 in the pichia pastoris is realized.

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.

The invention discloses a strain and method for increasing the yield of ergothioneine in saccharomyces cerevisiae and application, and belongs to the field of bioengineering. The invention relates to a method for improving the yield of ergothioneine in saccharomyces cerevisiae through a protein fusion technology, gene integration optimization and a culture medium strategy. A flexible linker FL1 is designed to fuse EGT1 and EGT2, the gene copy number and the SAM metabolic pathway are optimized, and a specific culture medium formula is combined, so that the shake flask fermentation yield reaches 122.5 mg / L, and the yield of a 10L fermentation tank reaches 2540 mg / L. The invention provides an efficient technical scheme for industrial production of ergothioneine.

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 for oil gas gene exploration and detection, and belongs to the technical field of intelligent oil gas exploration. The method solves the problem that an existing probe still has limitation due to the fact that the existing probe mainly depends on manual result interpretation and lacks data intelligent processing capacity, by introducing edge calculation and large model technologies, after gene sequence data in a soil sample is detected by the probe, quantitative analysis is carried out by utilizing a fluorescent quantitative PCR technology, and the accuracy of the gene sequence data in the soil sample is improved. The target gene is amplified by using the primer sequence to obtain a gene copy number, and the oil and gas reservoir probability of the soil sample is obtained; and the real-time gene sequence data and the oil and gas reservoir probability data are processed through a lightweight convolutional neural network to generate an oil and gas reservoir probability distribution thermodynamic diagram, so that cloud dependence is reduced, a probe can process the acquired data in real time on site, and a drilling target in a target area is quickly obtained according to the oil and gas reservoir probability distribution thermodynamic diagram. Therefore, the invalid drilling period is effectively shortened, the exploration cost is reduced, and the exploration efficiency is improved.

The present invention provides a recombinant microorganism for simultaneously producing sesquiterpenes and primary metabolites, wherein the primary metabolites include at least one of pantothenic acid, squalene, fatty acids and vitamin B2; the strain contains the mevalonate pathway, the β-farnesene synthesis gene and genes related to the synthesis of primary metabolites; the genes related to the mevalonate pathway include ERG10, ERG13, tHMG1, ERG12, ERG8, MVD1, IDI1 and ERG20 genes, and the copy numbers are ERG10:ERG13:tHMG1:ERG12:ERG8:MVD1:IDI1:ERG20 = 2:2:X:2:2:2:2:2, where X is an integer greater than or equal to 1; the β-farnesene synthesis gene includes bFS, and the sequence is as shown in SEQ ID NO.1. The present invention realizes high yield of primary metabolites while producing sesquiterpene compounds.

The present invention relates to a c-myc gene copy number microdroplet digital PCR detection kit, belonging to the field of biomedicine technology. The present invention provides a c-myc gene copy number microdroplet digital PCR quantitative detection method, which calculates the c-myc gene copy number based on the fluorescence values of the c-myc gene and an internal reference gene; the present invention discloses primers and probes for the above method and kit. By combining the present invention with clinical pathological information, follow-up results, treatment response, and fluorescence in situ hybridization methods, the optimal threshold value of c-myc gene copy number amplification variation is determined, which serves as a reference indicator for the prognosis of specific tumor patients, providing a convenient and reliable auxiliary diagnostic tool for clinical practice.

Disclosed in the present invention is a use of an MP1 gene copy number variation detection reagent in preparation of an NK / T cell lymphoma prognosis kit. The present invention further provides a detection kit comprising the LMP1 gene copy number variation detection reagent, a use thereof, and an NK / T cell lymphoma prognosis method. In the present invention, a copy number variation event of LMP1 gene in an EB virus genome is used as a novel prognosis marker, which has a good prediction value for the overall survival and progression-free survival prognosis of an NK / T cell lymphoma patient, and can realize more accurate and effective prognostic assessment for the NK / T cell lymphoma patient.

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 invention relates to a circular vector for plant transformation comprising: a first tethering nucleic acid and a second tethering nucleic acid; a nucleic acid encoding a polynucleotide of interest (POI); a nucleic acid comprising an origin of replication; and two or more nucleic acids encoding replicon proteins and methods for using the same. Also provided are plants produced by the methods of the invention and products produced from the plant.

The invention discloses a recombinant baculovirus transfer vector as well as a preparation method and application thereof. The recombinant baculovirus transfer vector comprises an early promoter and a late promoter which are arranged in sequence, and the downstream of the early promoter and the downstream of the late promoter are independently and operably connected with a porcine circovirus type 2 Cap protein coding frame. According to the recombinant baculovirus transfer vector, the copy number of exogenous genes of a traditional insect vector is greatly increased, the upper limit of the yield of the exogenous genes expressed by the vector is greatly improved, and by carefully designing the arrangement direction and sequence of coding frames of the recombinant baculovirus transfer vector, matching and combination of promoters in the morning and later periods and the like, the yield of the exogenous genes is improved. The high expression of the porcine circovirus PCV2 Cap protein in insect cells is realized.

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.

This invention is at least one panel of qPCR / RT-qPCR assays which enables simultaneous testing for the presence of multiple species and subgroups of cyanobacteria that produce microcystin, anatoxin, saxitoxin, and cylindrospermopsin cyanotoxins. The method takes into account that some cyanobacteria species may carry genes associated with multiple toxin types. Testing for each toxin type is conducted under standardized test conditions which allow quantification of the number of gene copies present for cyanotoxins which may contribute to the overall toxin level.

The invention particularly relates to a primer probe and a kit for detecting the copy number of a GJB2 gene of a non-syndromic deafness patient by using microdroplet digital PCR (Polymerase Chain Reaction), and application of the primer probe and the kit. The primer probe combination provided by the invention comprises a target gene detection primer pair, a target gene detection probe, an upstream primer of a reference gene, a downstream primer of the reference gene and a probe of the reference gene, and the sequence information of the primer probe combination is shown as SEQ ID NO.6-11; the invention also provides a kit for detecting the copy number of the GJB2 gene of a non-syndromic deafness patient. When the primer probe combination and the kit provided by the invention are applied to detection of copy number variation of the GJB2 gene of a patient with non-syndromic deafness, the accuracy of hereditary deafness detection is remarkably improved; whether GJB2 gene expression is abnormal or not is judged in an auxiliary mode according to the judgment result, disease prognosis is evaluated, potential treatment drugs are screened, genetic counseling services are provided or personalized medical schemes are formulated, and the method has wide application prospects.

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.

The present invention relates to primers, probes, kits and methods for detecting copy number variations of SMN1 and SMN2 genes. The kit of the present invention combines amplification refractory mutation system PCR (ARMS-PCR) and competitive primer PCR (COP-PCR), and is paired with the Taq-Man probe method, which can effectively distinguish the differences between the SMN1 gene and the SMN2 gene at two different sites in exons 7 and 8, while ensuring the amplification efficiency of the target, and has high sensitivity, good specificity and high stability (precision). In addition, the kit is paired with stable PCR amplification reaction components, which can effectively distinguish the differences between SMN1 deletion, carriage and normal, and can also calculate and analyze the copy number of SMN2, providing a basis for the disease analysis, medication guidance and prognosis of SMA patients.

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 provides a probe set for detecting the copy number of a TSPAN12 gene and application of the probe set. And the probe group comprises probes of No. 2 to No. 6 exons of the TSPAN12 gene. The probe with extremely high specificity is designed by avoiding a high GC content and a repetitive sequence region, so that a non-specific amplification phenomenon is effectively avoided, pathogenic variations such as point mutation, tiny insertion or deletion and the like of the TSPAN12 gene can be accurately detected, and the possibility of misdiagnosis and missed diagnosis is reduced. The method is especially suitable for TSPAN12 gene detection of familial exudative vitreoretinopathy (FEVR).

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.