58 results about "Polygenic Characters" patented technology

The invention relates to a primer pair and a probe used for noninvasive polygene methylation combination detection for early stage colorectal cancer, and includes the primer pair and the probe used for detecting methylation of genes Spetin9, NDRG4, BMP3, THBD and SDC2 and the primer pair and the probe for internal reference ACTB; the sequences of the primer pair and the probe are represented as the SEQ ID No.1 to the SEQ ID No.18. The invention also provides a kit containing the primer pair and the probe and applications thereof. The application method includes free DNA extraction from a plasma specimen, sulfite conversion, PCR amplification reaction, fluorescent signal detection and result determination. The kit and the method are suitable for methylation detection of the five genes Spetin9, THBD, SDC2, NDRG4 and BMP3 in human peripheral blood; compared with a conventional colorectal cancer diagnosis method, the application method fully utilizes the free DNA extraction from a plasma specimen, the DNA methylation and QPCR associated technologies, thus developing the kit having high sensitivity and specificity. The primer pair, probe and kit are used for performing early stage noninvasive screening to human colorectal cancer.

The invention provides a systemic detection method of a tumor marker and application thereof. The method comprises the following steps: 1) providing a sample from a checking object, wherein the checking object is a person; 2) detecting the sample of step 1), wherein in the detection step, whether various protein tumor markers exist or not is determined by utilizing high-throughput nucleic acid sequencing and determination comprising polygene mutation, polygene methylation, a plurality of micro RNAs (Ribonucleic Acid) and the like and utilizing a protein chip. According to the systemic detection method of the tumor marker and the application thereof, the different tumor markers are subjected to systemic detection including DNA (Deoxyribonucleic Acid), RNA, protein and the like, so that thesensitivity of early-stage detection of the tumors is improved.

The present invention is directed to the identification of genostatic factors, methods of determining the association of a plurality of genes with polygenic disorders, and method of assessing the sensitivity and specificity of the risk of polygenic disorders. In particular, the present invention discovers that the association between a polygenic disorder phenotype and polygenes may be masked. Incorporating genostatic factor, such as maternal age, birth disorder, the androgen receptor gene, gender, and age, into the statistical analysis of the association between phenotypes and genotypes reveals statistically significant relationship between the two. Accordingly, the present invention provides novel methods in determining the association of a plurality of genes with a polygenic disease and the likelihood of having the polygenic disease.

The invention relates to a principle of quantitative trait locus (QTL) positioning. The QTL refers to the position of a gene for controlling quantitative traits in a genome; A polygenic system for controlling the quantitative trait is used as integration for research on the basis of classical quantitative genetics of polygene hypothesis by several genetic designs and statistical models; and the genetic characteristics of the quantitative traits are described by genetic parameters, such as hereditary capacity and genetic variance.

The invention discloses a polygene assembling carrier system and its polygene assembling method. The system comprises a 1-type accepting carrier, a 2-type supply carrier, and a supply plasmid capable of being selectively used and generating self-deletion of a selection marker. The system applies a Cre / loxP recombination system and irreversible recombination sites of two groups of mutant type loxP; under the function of Cre enzyme, the supply carrier is integrated to the accepting carrier through the recombination of a wild loxP site; then a framework of the supply carrier is automatically deleted through the irreversible recombination of one group of mutant type loxP, and only the target gene is left on the accepting carrier; by repeating the alternation of the 2-type supply carrier and the assembly of the accepting carrier, the rapid assembly of the polygene can be realized. The invention is the simplest and most high-efficient polygene carrier system at present. Therefore, the polygene assembling carrier system provides an operable technology platform for important and complex biosynthetic pathways and important agronomic trait polygenic inheritance engineering; the polygene assembling carrier system is of important application value.

The invention relates to a chloroplast simultaneity expression seven exogenous genes vector forming method. It belongs to genetic engineering technique field. It includes the following steps: forming a chloroplast expression vector containing faeC, faeD, faeE, faeF, faeG, faeH, and faeI seven gene orders which form K88ac flagellin gene cluster; extracting coding K88ac wild bacterium C83907 plasmid; using primer 1 whose nucleotide sequence is same with SEQ ID NO: 1 and primer 2 whose same with SEQ ID NO: 2 to process PCR amplification; nucleotide sequence 3018bp is same with SEQ ID NO.5; using primer 3 whose same with SEQ ID NO:3 and primer 4 whose same with SEQ ID NO:4 to process PCR amplification; the nucleotide sequence is same with SEQ ID NO.6; connecting with pMD18-T vector; transforming to escherichia coli DH5 alpha; and gaining escherichia coli clone pTEI with faeCD sequence. The invention overcomes the key defect of polygene technique of plant genetic improvement, and offers a good method of producing effective antibody plant bacterin for baby pig.

The invention discloses a plasmid vector pSilencer-U6 / H1-(shRNA)n which can be used for simultaneously expressing a plurality of shRNA, a construction method and application thereof. The vector is composed of two groups of sequences of multiple cloning sites (MCS), a plurality of target gene shRNA transcription units and plasmid sequences. The shRNA transcription units of a plurality of different or same target genes can be connected in series between the two groups of the sequences of the multiple cloning sites of the vector, a plurality of shRNA transcription units thereon can express one target gene for many times or express a plurality of different sites of one target gene or express different target genes after the transcription of cells or animals, so as to specifically inhibit the expressions of two or more target genes. The vector can be easily used in the gene function research and the research and development of the gene therapeutic drugs for diseases.

The invention discloses a lung adenocarcinoma individuation prognosis evaluation method based on a polygene expression characteristic spectrum. The method comprises the following steps that a lung adenocarcinoma prognosis risk gene list and weight are acquired; a prognosis evaluation model is established by using a tumor tissue transcriptome and survival data of a patient with lung adenocarcinoma;a risk score of the patient is calculated according to a gene expression spectrum of the tumor tissue of the patient with lung adenocarcinoma; the annual survival probability of the patient is calculated according to the risk score of the patient. According to the method, the annual survival probability of the patient with lung adenocarcinoma and the actual annual survival probability are highlyidentical, the linear correlation R2 is equal to 0.994, and the P value is equal to 2.86E-43. It is proved that the method has high prediction accuracy, and the state is highly identical to the actualsurvival state. Meanwhile, for each patient with lung adenocarcinoma, the method can provide a specific survival probability curve of the patient.

The invention discloses a polygene multi-target enrichment method. A capture probe library is designed, and target gene sequences are enriched in a probe capture way at a DNA (deoxyribonucleic acid) level, so that detection on all variation types, such as mutation, deletion, insertion, fusion and copy number amplification, of target genes is realized, and the defect that presently the target gene sequences are enriched by a PCR method at the DNA level, only mutation and deletion of the target genes can be detected, and the fusion cannot be detected is overcome.

The invention discloses hansenula polymorpha with a double selection marker and an application thereof. The hansenula polymorpha with a double selection marker provided by the invention is uracil-synthesized and tryptophan-synthesized double-deficient hansenula polymorpha HUT-31 with a preservation number of CGMCC No.4778. The strain has a uracil and tryptophan double-auxotrophic selection marker, maintains the physiological biochemical characteristics of wild-type strains, facilitates the culture of recombinant strains and the high-level expression of heterologous protein; not only recombinant strains can be selected easily and rapidly, but also the double selection marker can increase the copy number of target genes integrated on a chromosome, can increase the expression level of targetprotein, and has important industrial application value. Meanwhile, the double selection marker is also applicable to the polygene genetic modification of hansenula polymorpha in functional genomics,synthetic biology, and metabolic engineering.

The invention provides a floating population information analysis and warning and alarming method based on biological instructive calculation. Wherein, with current floating population information system from department of public safety, extracting need information to calculate floating population data with the multilayer chromosome gene expression programming algorithm and overlap-expressive polygene evolution algorithm in biological instructive calculation filed; and completing data mine and classification and isolated-point analysis to realize the warning and alarm. This invention can take deep analysis to trace and trend of abnormal floating population and different criminal feature, builds a intelligent analysis platform, and can also provide useful information for local peace.

The invention discloses a rapid detection kit and a detection method of a v.parahaemolyticus multiple virulence factor GeXP. The method comprises the following steps of: extracting a genome DNA (Deoxyribose Nucleic Acid) of the v.parahaemolyticus; performing multiple PCR (Polymerase Chain Reaction) amplification on 13 pairs of polygene PCR primers and a pair of general primers by taking the genome DNA as a template to obtain a multiple PCR reaction product; and detecting and analyzing by using a GenomeLab Gexp genetic analysis system. In the invention, a specific primer is designed specific to 13 kinds of known virulence genes of v.parahaemolyticus, and 13 kinds of genes are detected rapidly and efficiently at one time through a single-tube reaction to detect whether a sample contains v.parahaemolyticus which carries virulence genes and has potential pathogenic ability can be known, so that the detection coverage is wide, the accuracy of pathogenicity evaluation of v.parahaemolyticus can be greatly increased, and a more effective detection measure is provided for pathogenicity evaluation of v.parahaemolyticus. The method has important application prospects on the aspects of aquaculture disease detection, epidemiological investigation, pathogenic bacterium detection in foods, aquatic product import and export detection, hospital infection monitoring, and the like.

The invention discloses a chip and method for real-time PCR (polymerase chain reaction) gene detection at a polygenic mutation site. The gene detection chip comprises a porous PCR plate, wherein a PCR reaction system is arranged in each pore of the porous PCR plate; the PCR reaction system comprises a plurality of primer pairs for the specific amplification of target gene targeting sequences, fluorescent probes, PCR buffer solutions, dNTPs (deoxyribonucleotide triphosphates) and a GoldTaq enzyme; each primer pair contains a continuous nucleotide sequence composed of at least 15 continuous nucleotides in target gene exons; and fluorescent molecules of the fluorescent probes emit fluorescent signals in the PCR amplification process. The detection rate of the method on big genes can be up to 98% or above.

The present invention relates to a recombinant polynucleotide encoding a polygene coding for at least three polypeptides wherein at least one of the genes constituting the polygene is of non-viral origin, at least two of the polypeptides encoded by the genes constituting the polygene are each capable of at least transiently interacting with at least one other polypeptide encoded by a gene of said polygene, and the genes constituting the polygene are each connected to one another by a sequence coding for at least one protease cleavage site. The present invention also relates to polyproteins encoded by the polygene. Further embodiments of the present invention are a vector containing the recombinant polypeptide, a host cell containing the recombinant polypeptide and / or the vector and a non-human transgenic animal transformed with the recombinant polypeptide and / or the vector. The present invention also relates to methods for the production of the polynucleotide and for the manufacture of multiprotein complexes. The embodiments of the present invention are particularly useful in gene therapy, drug candidate screening, vaccine production and crystallisation of multiprotein complexes for structural investigations.

The invention discloses a polygene joint detection probe for early diagnosis of tumour, a primer and a kit. The probe and the primer contain SEQ ID NO: 1-SEQ ID NO: 91, can be used for joint detection of p53, KRAS, EGFR, BRAF, PIK3CA and MEK1 genes; can be used to simultaneously detect a plurality of driving abrupt changes in one PCR reaction tube; has high sensitivity and can be used to detect 5-10 copy abrupt change; has strong singularity without generation of non-specificity signals from 10 ng of wild type genome DNA; and has advantages of rapid detection speed, easy operation and low cost.

The invention discloses a new wheat stripe rust resisting polygene-polymerized flower-nurture breeding method. Wheat-breeding parent materials with excellent agronomic characters are selected; 3 major micro-species of puccinia striiformis, which are recently popular, are used to evaluate stripe rust resistance of the wheat-breeding parent materials at seedling stage and adult stage; 3 wheat-breeding parent materials, which have high-resistance immunity to major popular individual micro-species of various puccinia striiformis, are sieved; the 3 wheat-breeding parent materials are hybridized and duplex hybridized to obtain duplex hybridized polygene-polymerized subsequent generation; the duplex hybridized subsequent generation is then cultured and wheat anther tissue is inoculated to one-step seeding culture medium of wheat anther, thereby obtaining complete pollen plant; chromosome doubling liquid with colchicines and dimethyl sulfoxide is injected to tillering node, thereby carrying out pollen plant chromosome doubling to obtain doubly purified individuals of doubled haploid which can be self-hybridized to form a stable doubled haploid system; and stripe rust resisting individual micro-species validation is carried out on the doubled haploid system, so as to obtain a new stripe rust resisting type or species system which is polymerized with 3 stripe rust resisting genes.

The invention discloses a construction method of a gene mutation detection library for tumor treatment cardiac toxicity prediction. The construction method is characterized by covering totally 56 genetic variation loci on 38 related genes of human tumor treatment cardiac toxicity. The construction method of the invention quickly completes the construction of the library by aiming at separately managing a plurality of target sequences, the construction process of the entire library only needs 2 to 3 hours, and only 45 min manually, so that the difficulty of detecting somatic cell polygenes and multiple target points required by predicting whether a greater cardiac toxicity is produced or not before a patient receives a tumor drug therapy clinically at present can be solved very effectively, and the cost is low.

The invention belongs to the technical field of genetic engineering vaccines, and particularly relates to an African swine fever virus tandem gene, a coexpression vector, a construction method and application. The recombinant vector is obtained through the steps that an artificially synthesized polygene clone expression element is connected to the portion between the BglII and BamHI restriction enzyme cutting sites in a pIRES2-egfp vector, and oriR101 is inserted in the portion behind the XbaI site of 1986 nt of the pIRES2-egfp vector, wherein the nucleotide sequence of the gene clone expression element is shown in SEQ ID NO: 1; the recombinant vector is transformed into escherichia coli competent cells, screening and cultivation enlargement are conducted, then polygenic protein is collected and purified, and a subunit vaccine against the African swine fever disease is obtained; the recombinant vector is transformed into lactobacillus competent cells, and after screening and cultivation enlargement are conducted, a lactobacillus vaccine is obtained. The prepared vaccine stimulates the body to generate an antibody in different levels and different gradations, stimulates the body toproduce a strong and effective specific immune response, exerts high-efficiency immune protection, and makes an immune response against the African swine fever virus.

The invention discloses an analysis method for predicting body weight increase caused by treating schizophrenia through a second-generation antipsychotic based on the polygene combination interactiveeffect. The method comprises the steps that a sample is prepared for collecting peripheral blood of a patient with body weight increase caused by treating schizophrenia through the second-generation antipsychotic, a hypotonic salt fractionation method is used for extracting a genome DNA sample; a MODLI-TOF flight mass spectrum detecting method is used for performing genetic typing of a 5-HT2CR gene, a histamine 1 receptor gene, an oxytocin gene, an NPY / R gene, a Leptin gene, an adiponectin gene, an FGF21 gene and a FGF23 gene; data analysis and extraction are performed, instrument original data is aligned, data with no noise interference for statistic analysis is obtained, a generalized multi-factor dimension reduction method is used for performing the quantitative trait gene-gene interaction effect, crossed verification is further applied, and the gene interaction effect is used for predicting body weight increase. By adopting the generalized multi-factor dimension reduction method, the continuous ending variable is processed, covariance is introduced, and the method has the advantages of greatly enlarging the application range and greatly increasing the prediction accuracy.

The invention discloses a breeding method of a high oil content disease-resistant rapeseed variety. The method comprises inducing male sterility of an excellent variety through a chemical hybridizing agent SX-1, carrying out hybridization, multiple crossing and backcrossing on various varieties to obtain a multigene aggregation group, carrying out disease resistance identification and apparent character selection on the multigene aggregation group to obtain a single plant having excellent integrated properties, carrying out microspore culture on the single plant, carrying out disease resistance identification on the obtained double haploid segregation population, carrying out different-area allopatric generation adding and shuttle breeding to obtain a variety Lxn having high oil content, low pole height and powdery mildew and downy mildew resistance. Compared with the traditional rapeseed polygene pyramiding technology, the breeding method has high efficiency, easily realizes synchronous pyramiding of multiple excellent genes and improvement of properties, easily realizes massive hybridization in gene pyramiding selection, improves population selection pressure and probability and significantly shortens a effective selection cycle.

The invention provides a reliable method for detecting H1N1 influenza A virus variant by a multiple target point gene and a kit. Groups of primer needles are designed to multiple gene target points. The accuracy of each fragment amplified is intuitively determined by displaying size of PCR (Polymerase Chain Reaction) sequence fragment through capillary electrophoresis fluorescent detection. The polygene target points confirm positive results layer by layer, so that the specificity of detection result is ensured. In addition, the kit can detect non-H1N1 influenza A injection and non A influenza injection samples, so as to further provide associated information for diagnosis.

The invention discloses a polygene sequencing primer and a detecting method for detecting congenital hypothyroidism, belonging to the fields of molecular biology and clinical examination. A group of polygene PCR primer sequences for detecting congenital hypothyroidism is composed of one or more primer sequences selected from a PCR primer sequence of a Pax-8 gene, a PCR primer sequence of a TG gene, a PCR primer sequence of an IYD gene, a PCR primer sequence of a DUOX2 gene, a PCR primer sequence of an NIS gene, a PCR primer sequence of a TSHR gene and a PCR primer sequence of a TPO gene. The polygene sequencing primer has the advantages that a plurality of disease-causing genes capable of inducing congenital hypothyroidism are simultaneously detected through new generation sequencing, so that the polygene sequencing primer is low in time consumption, high in detecting efficiency, low in cost, relatively high in sensitivity and specificity and capable of rapidly and comprehensively realizing gene mutation detection and disease classification of clinical patients.

The invention discloses a tri-cistron muscle specific two-way co-expression gene transferant and a preparation method thereof. The gene transferant comprises an MARs cow matrix attachment region, short hairpin RNA (shRNA) taking the third exon (926-947 loca) of cow muscle colyone as a target spot, an optimally synthesized muscle specific promoter (MRF4), a humanization n-3 desaturase gene (FAD3), an internal ribosome entry site (IRES), a red fluorescent protein gene (DsRed) and a termination signal region (SV40polyA), and the independent gene transferant does not have a pronucleus main vector sequence. According to the tri-cistron muscle specific two-way co-expression gene transferant and the preparation method, an RNA interference technology is firstly combined with the muscle specific overexpression FAD3 and DsRed, therefore, gene knock-down and inverse overexpression of related genes can be performed simultaneously, and a novel thought and approach can be supplied for safely and efficiently improving characters, such as muscle and disease related characters, controlled by polygenes.

The invention relates to a method for constructing a BmNPV-domestic silkworm larva polygene expression system, which comprises the following steps: (1) an I-SceI expression cassette controlled by arabinose promoter (pBAD) is recombined into the genome of E.coli D H10B to obtain new recipient bacteria I-SceI DH10B; (2) a donor plasmid used for combined transformation is constructed; (3) a recombination site of cre / loxp replaces v-cath gene and chitin gene in the BmBacmid genome, and two I-SceI sites replace p70 gene in the BmBacmid to obtain a receptor I-sceI BmMultibac; (4) an exogenous gene is induced by the three methods of cre-loxp recombination, Tn7 recombination and combined transformation; (5) a recombined Bacmid DNA is prepared by a conventional method, and is used for transfecting insect cells or injected into the silkworm larva. The BmNPV-domestic silkworm larva polygene expression system lays a foundation for studying multimeric proteins, the interaction between proteins and preparing virus-like particles.

The invention discloses a plant polygene expression vector composition and application thereof. The vector composition consists of a vector A and a vector B; the nucleotide sequence of the vector A comprises a T-DNA terminal sequence, a multiple cloning site sequence and a specific recombinant site sequence sequentially connected in series; and the nucleotide sequence of the vector B comprises a specific recombinant site sequence, a multiple cloning site sequence and a T-DNA terminal sequence sequentially connected in series. The plant polygene expression vector composition is reduced to minimum, and copy number, stability and conversion efficiency of plasmids are improved. Under the condition of ensuring that the conversion efficiency is not remarkably reduced, the smaller the plasmids are, the longer the fragment of the containable exogenous DNA is, so the vector composition is very favorable for experiment operation.

The invention discloses a molecular marker and application of grape anthracnose resistance main effect QTL (quantitative trait locus), and relates to the field of genetics and breeding of fruit plants. The quantitative trait locus determining grape anthracnose resistance is subjected to QTL positioning for the first time, main-effect QTL locus RA of grape anthracnose resistance is determined, rate of contribution of the locus is 71.5%, and 37.7% of grape anthracnose resistance characteristics can be explained. Accuracy in target trait judgement is improved through the linked molecular marker M1E12-1500-m screened based on the locus, and a foundation is laid for improvement of polygene controlled trait based on molecular breeding. In addition, the molecular marker M1E12-1500-m is linked with the main-effect QTL locus RA at a distance of 0.96cM, and the molecular marker M1E12-1500-m is closely linked with the target locus. Therefore, the obtained marker has good application value, accuracy and efficiency in molecular marker-assisted selection can be improved, and progress of improvement in anthracnose resistance of grapes is accelerated.

The invention belongs to the field of bioengineering and provides a transgenic method for increasing oil peony seed oil content. The method includes: constructing a polygene plant expression vector; transferring into a peony plant tissue through agrobacterium tumefaciens to realize expression and heredity, wherein construction of the polygene plant expression vector includes that a polycloning site is introduced at a 3' terminal of a 35S promoter on Pbi1211-GUS to construct a vector Pbi121-35S MCS, and exogenous genes WRI1, LEC2 and FAD2 are sequentially inserted into Pbi121-35S MCS to form the polygene plant expression vector Pbi121-WRI1-LEC2-FAD2. On the basis of the constructed polygene plant expression vector, effective expression in peony plants can be realized, and harvested peony seeds can be stable in inheritance and high in oil content.

An efficient plant polygenic expression carrier pBtslm / aroM12 prepared from basic carrier pCAMBIA contains the antibiotic resistance gene NPTII, the glyphosate resistant gene aroAM12 and pest-resistant gene BtSlm. Said expression carrier can be used to transform colibacillus to obtain its colibacillus transformant, or to transform Agrobacterium to obtain its Agrobacterium transformant. Said transformat can be used to transform plant, obtaining the plant resisting glyphosate and pests.

The invention discloses a plant polygene genetic transformation method, and belongs to the technical field of plant biological engineering. The method comprises establishing a single-genotype high-quality high-agrobacterium-infection-rate embryonic callus line; and infecting the above callus line by using grobacterium with different binary vectors, and performing double-antibody screening to obtain a transgenic positive plant containing multiple different exogenous genes. The method is mainly used to solve the problem that plant is difficult for genetic transformation and multiple-property comprehensive improvement. Experiment data show that the method can help to successfully obtain the positive transgenic plant possessing the single genotype background within 4-5 months, the genetic transformation efficiency is up to 60%, the target of converting 200 gene vectors per person per year can be realized, and the working efficiency of generating 3000 independent positive transgenic plants containing multiple different exogenous target genes can be provided.