271 results about "Karyotype" patented technology

Methods for producing a paired tag from a nucleic acid sequence are provided in which the paired tag comprises the 5′ end tag and 3′ end tag of the nucleic acid sequence. In one embodiment, the nucleic acid sequence comprises two restriction endonuclease recognition sites specific for a restriction endonuclease that cleaves the nucleic acid sequence distally to the restriction endonuclease recognition sites. In another embodiment, the nucleic acid sequence further comprises restriction endonuclease recognition sites specific for a rare cutting restriction endonuclease. Methods of using paired tags are also provided. In one embodiment, paired tags are used to characterize a nucleic acid sequence. In a particular embodiment, the nucleic acid sequence is a genome. In one embodiment, the characterization of a nucleic acid sequence is karyotyping. Alternatively, in another embodiment, the characterization of a nucleic acid sequence is mapping of the sequence. In a further embodiment, a method is provided for identifying nucleic acid sequences that encode at least two interacting proteins.

A source of multipotent amniotic fluid / fetal stem cells (MAFSCs) is disclosed. MAFSC are of fetal origin and have a normal diploid karyotype. These cells are characterized by the following cell surface markers: SSEA3, SSEA4, Tra-1-60, Tra-1-81, Tra-2-54, HLA class I, CD13, CD44, CD49b, CD105 and are distinguished by the absence of the antigen markers CD34, CD45, and HLA Class II, but are distinguished from mouse embryonic stem cells in that these cells do not express the cell surface marker SSEA1. MAFSC express the stem cell transcription factor Oct-4. MAFSC cells can be propagated for an indefinite period of time in continuous culture in an undifferentiated state. The MAFSCs have the ability to differentiate in culture in a regulated manner, into three or more subphenotypes. Cells can then be differentiated into endodermal, mesodermal and ectodermal derived tissues in vitro and in vivo. A method for isolating, identifying, expanding and differentiating MAFSCs is disclosed.

A new method for genomic analysis, termed “Sequence-Based Karyotyping,” is described. Sequence-Based Karyotyping methods for the detection of genomic abnormalities, for diagnosis of hereditary disease, or for diagnosis of spontaneous genomic mutations are also described.

Alterations in the genetic content of a cell underlie many human diseases, including cancers. A method called Digital Karyotyping provides quantitative analysis of DNA copy number at high resolution. This approach involves the isolation and enumeration of short sequence tags from specific genomic loci. Analysis of human cancer cells using this method identified gross chromosomal changes as well as amplifications and deletions, including regions not previously known to be altered. Foreign DNA sequences not present in the normal human genome could also be readily identified. Digital Karyotyping provides a broadly applicable means for systematic detection of DNA copy number changes on a genomic scale.

Existence of human trophoblast stem (hTS) cells has been suspected but unproved. The isolation of hTS cells is reported in the early stage of chorionic villi by expressions of FGF4, FGFR-2, Oct4, Thy-1, and stage-specific embryonic antigens distributed in different compartments of the cell. hTS cells are able to derive into specific cell phenotypes of the three primitive embryonic layers, produce chimeric reactions in mice, and retain a normal karyotype and telomere length. In hTS cells, Oct4 and fgfr-2 expressions can be knockdown by bFGF. These facts suggest that differentiation of the hTS cells play an important role in implantation and placentation. hTS cells could be apply to human cell differentiation and for gene and cell-based therapies.

Existence of human trophoblast stem (hTS) cells has been suspected but unproved. The isolation of hTS cells is reported in the early stage of chorionic villi by expressions of FGF4, fgfr-2, Oct4, Thy-1, and stage-specific embryonic antigens distributed in different compartments of the cell. hTS cells are able to derive into specific cell phenotypes of the three primitive embryonic layers, produce chimeric reactions in mice, and retain a normal karyotype and telomere length. In hTS cells, Oct4 and fgfr-2 expressions can be knockdown by bFGF. These facts suggest that differentiation of the hTS cells play an important role in implantation and placentation. hTS cells could be apply to human cell differentiation and for gene and cell-based therapies.

Non-embryonic blastomere-like totipotent stem cells are disclosed. Most preferably, such cells are obtained from various tissues of postnatal mammals (e.g., using tissue biopsied from the mammal), are smaller than 1 μm, have normal karyotype, and do not spontaneously differentiate in serum-free medium without differentiation inhibitors. These non-embryonic blastomere-like totipotent stem cells typically express CD66e, CEA-CAM-1 and telomerase, but do not typically express CD10, SSEA-1, SSEA-3, and SSEA-4. Such blastomere-like totipotent cells can be differentiated into ectodermal, mesodermal, or endodermal tissues, including placental tissues and germ cells. Moreover, when implanted into a mammal, such cells will not be teratogenic.

The invention provides a detection method for genomic copy number variation and a device comprising the method. The method comprises the following steps: inputting original data, cleaning quality control, aligning sequences to a reference genome, calculating a unique alignment sequence number by using windows of different sizes, performing GC correction and reference correction, shielding undetectable areas, segmenting CBS, integrating karyotype reports and generating reports, a complete set of the detection methods and devices is established through experimental exploration and optimization,through successive application of the specific sequential steps, the step of the reference correction is creatively adopted, and the windows with different sizes are used for performing alignment andintegration, the steps cooperate mutually, and finally the sensitivity and specificity are improved, so that the detection accuracy and result forms can conform to clinical demands, and the detectionmethod for the genomic copy number variation and the device comprising the method are high in automation degree, easy to expand, high in detection accuracy, capable of lowering the cost of data analysis, and extremely high in application value.

The invention discloses peanut oligonucleotide probes and their design method and use method. The peanut oligonucleotide probes comprise eight probes having nucleotide sequences shown in the formulas of SEQ ID NO. 1 to NO. 8. The microsatellite and telomere sequences are used for developing the oligonucleotide probes, the novel oligonucleotide probes are combined into a probe set and oligonucleotide types of the peanut cultivar and wild peanut are constructed, an economic, efficient and high universality peanut cytological marker design technique and chromosome recognition technique are built, peanut chromosome markers are enriched, genomes and chromosomes of the peanut cultivar and wild peanut are identified, and chromosomal structural variation of wild peanut species is identified. Through use of a high-throughput small data simplified sequencing (for only measuring the amount of genomic 4Gb data) and bioinformatics analysis, peanut oligonucleotide probe markers are successfully developed. The invention provides a novel effective method for low-cost and high-efficiency development of peanut cytological markers.

The invention provides a method for producing porcine circovirus type II capsid protein by utilizing a silkworm bioreactor, and belongs to the field of biotechnology. In the method, by taking a bombyx nuclear polyhedrosis virus as a vector, porcine circovirus type II capsid protein genes are integrated into a polyhedrosis promoter of the bombyx nuclear polyhedrosis virus so as to express target protein by a mode of the homologous recombination of a homologous arm of the nuclear polyhedrosis virus in a transfer vector and bombyx nuclear polyhedrosis virus (BmNPV) genes; a recombinant bombyx nuclear polyhedrosis virus containing target protein genes is obtained by a plaque sieving method, and the target protein is expressed in large scale by using a bombyx bioreactor so as to prepare a subunit vaccine containing recombinant porcine circovirus type II capsid protein; and piglet infection experiments verify that the subunit vaccine has the excellent immune protective effect. The method has the characteristics of high expression efficiency, good activity of the target protein, low production cost and the like, and is suitable for large-scale production.

The invention provides a method for preparing chorionic mesenchymal stem cells. The method comprises the following steps: performing aseptic collection of placentas; separating chorionic tissues; acquiring chorionic mesenchymal stem cells; culturing the chorionic mesenchymal stem cells; performing frozen preservation on the chorionic mesenchymal stem cells; and reviving the chorionic mesenchymal stem cells. The method provided by the invention has the following technical effects: by adopting a pollution prevention method to reduce the probability of pollution from sources, the probability of pollution is effectively reduced by flushing a surface for multiple times; the pollution of hybrid cells is reduced; a single enzyme is adopted for digestion, thereby simplifying the process; the use of animal source components is reduced by using a serum-free culture medium for culture, the cell performance is stable, the in vitro long-term cultivation of the chorionic mesenchymal stem cells can be maintained, the form, proliferation ability, MSC surface marker expression and differentiation ability of the cells are maintained, and the expression of telomerase, the stable expression of cancer genes and the stable karyotype are maintained; and the enzyme digestion, frozen preservation and reviving cause no damage to the cells.

The invention relates to application of a fluorescent in-situ hybridization polyclonal separating probe for renal carcinoma and a kit thereof. Clonal fragments used by the polyclonal separating probe provided by the invention are respectively a combination of RP11-416B14, CTD-2522M13 and CTD-2516D6, and a combination of CTD-2312C1, CTD-2248C21 and RP11-959H17. According to the invention, the defect that karyotype analysis can not be performed conventionally after tumor cells are subjected to cell culture until a mitotic phase in the prior art is overcome. The typical gene modification in Xp11.2 translocation / TFE3 gene fusion related renal carcinoma is detected by using the FISH technology, thereby diagnosing the tumor. The fluorescent in-situ hybridization polyclonal separating probe application is high in accuracy, high in specificity, high in success rate, strong in fluorescent signals and convenient to operate. The invention can be applied in paraffin sections, widens the scope of detection specimens, and greatly optimizes the diagnosis method of Xp11.2 translocation / TFE3 gene fusion related renal carcinoma.

The invention discloses a quality control material of a chromosome aneuploid (T21, T18 and T13) detection kit. The quality control material comprises a positive reference material and a negative reference material. The positive reference material is a reference material set composed of a solution P21, a solution P18 and a solution P13, wherein the solution P21 is composed of solute and solvent, the solution P18 is composed of solute and solvent, the solution P13 is composed of solute and solvent, and the solvent of the solution P21, the solvent of the solution P18 and the solvent of the solution P13 are normal solutions. The normal solutions are in-vitro plasma of normal non-pregnant women of which the chromosome karyotype is 46, XX according to the analysis result. The solute of the solution P21 is PL21, the solute of the solution P18 is PL18, and the solute of the solution P13 is PL13. The quality control material is economical, simple and convenient to use and effectively achieves quality verification and performance evaluation of the chromosome aneuploid (T21, T18 and T13) detection kit.

The invention relates to a human parthenogenetic embryo stem cell line with two active X chromosomes and derivatives thereof. A karyotype of the human parthenogenetic embryo stem cell line is 46, XX before 10 generations; the karyotype of the cell line becomes a chimera from the 20th generation; all cells of the cell line lose an active X chromosome at the 35th generation; and in the inactivation detection of the X chromosome, only the state of the active X chromosome is maintained all along, but the presence of the inactivated X chromosome is not discovered. The parthenogenetic activation can be achieved by adopting an artificial activation condition causing a second polar body to be discharged; therefore, the human parthenogenetic embryo stem cell line and the derivatives thereof can be obtained. The cell line and the derivatives thereof have important significance on researching the characteristics of the cell on genetics and epigenetics and further applying the cell to clinically treating certain diseases; and a cell source for cell replacement therapy can be obtained by transforming genes of the cell or modifying the genes on the epigenetics.

The invention discloses a bombyx mori nuclear polyhydrosis virus (BmNPV) 39k inducible promoter and application thereof. A nucleotide sequence of the promoter is shown as SEQ ID No.1; the promoter has obvious BmNPV inducible promotion activity and can make cells start foreign gene expression when the cells are subjected to the BmNPV infection; an RNAi vector using a BmNPV multiplication essential gene as a target can be built by utilizing the promoter; the abundant expression of shRNA can be performed when the bombyx mori is subjected to the BmNPV infection; the shRNA is cut into siRNA in a cell by an enzyme; specific degradation is performed on the BmNPV multiplication essential gene mRNA to initiate the BmNPV multiplication essential gene to be transcribed and then silenced, so the aim of inhibiting the virus multiplication is fulfilled, and the promoter can be used for preventing and controlling the BmNPV infection of the bombyx mori, performing genetic engineering breeding of bombyx mori anti-BmNPV strains and providing a good reference mode for transgenic therapy of other biologic virus diseases.

The invention discloses a preparation method of recombinant nuclear polyhedrosis virus (NPV) which infects ectropis oblique. The preparation method comprises steps that: (1) recombinant vectors pFastBacDual-polh-helicase are prepared; (2) the recombinant vectors pFastBacDual-polh-helicase are transformed into E.coli DH10 BmBacmid competent cells; the cells are cultivated on a cultivating plate; positive plaques are obtained through blue-white selection; and recombinant BmBacmid-polh-helicase DNA is extracted from the positive plaques; (3) the ecombinant BmBacmid-polh-helicase DNA is transfected to domestic silkworm cultivated cells; cultivated cell supernatant is collected; and recombinant virus BmEoNPV is obtained; (4) the recombinant virus BmEoNPV is used for injection; and recombinant virus polyhedron is obtained. The recombinant nuclear polyhedrosis virus is sprayed on tea tree leaves, such that the ectropis oblique can be prevented from harming the tea trees. The preparation method also has advantages of low cost and controllable productions.

Provided is a method of preparing platinum nano particles by using cotton bollworm karyotype polyhedrin. Inherent structural features of the cotton bollworm karyotype polyhedrin are used, a PtC14 solution is added to a purified cotton bollworm karyotype polyhedral body solution, and the precious metal platinum nano particles which are even in distribution and consistent in diameter of the particles are obtained on the surface of the cotton bollworm karyotype polyhedrin through hatch, ultrasonic wave assistance reduction treatment. Due to the fact that a cotton bollworm karyotype polyhedron carrier has natural specific structures of functional group repetitive distribution, the complex preparation technology of a conventional carrier is avoided, and therefore production cost is reduced. The sizes and distribution of the precious metal platinum nano particles are effectively controlled through control of the alkaline hydrolysis degree of the cotton bollworm karyotype polyhedron, and the method of preparing the platinum nano particles by using the cotton bollworm karyotype polyhedrin can be significantly applied to catalytic fuel cells or biosensing or the like.

The invention discloses a construction method of a tilapia brain cell line. The method comprises the following steps: disinfecting fresh and live tilapia, taking brain tissue, cutting the tilapia intominced blocks, treating the minced blocks with collagenase, and adding trypsin; performing full suspension with a culture solution, and performing primary culture; when the bottom of a bottle is fullof cells, performing digestion with pancreatin-EDTA, and performing inoculation for secondary culture; and after 60 generations of secondary culture, adding colchicine in the logarithmic growth phasefor treatment, and performing karyotype analysis. The preparation method of the tilapia brain cell line, disclosed by the invention, is simple and convenient; and the tilapia brain cell line preparedby the method can perform secondary culture for a long time, is very sensitive to hemorrhagic disease viruses of tilapia, and can be widely applied to experiments.

The invention discloses a highly metastatic model of human melanoma, a highly metastatic cell subline of the human melanoma, creation methods for the highly metastatic model and the highly metastatic cell subline, and the dynamic detection of metastasis. The subcutaneously-transplanted mouse highly metastatic model and the corresponding cell subline are established in an in-vivo screening way in a mouse with severe combined immune deficiency (SCID) by using mouse lung metastasis, namely human malignant melanoma cell strain A375 pulmonary metastasis, wherein the highly metastatic cell subline of the human melanoma is A375sci, and has a human tumor cell karyotype; and 60 to 75 hypo-triploid-dominated chromosomes are acrocentric and have a heteroploid karyotype. The cell subline has the two routes of metastasis of blood trails and lymph. The in-vivo screening of the highly-metastatic model is performed by using animals with severe immune deficiency, and is expressed and applied in nude mice. A method for detecting Alu genes by using a polymerase chain reaction (PCR) method is simple, highly sensitive and highly specific, and can be used for detecting organ metastasis, particularly micrometastasis, in a human tumor animal-xenotransplantation model.

The invention discloses a kit and a method for cryopreserving induced pluripotent stem cells. The kit comprises: a first refrigeration working solution containing a first induced pluripotent stem cell medium, glycol and dimethyl sulfoxide (DMSO); and a second refrigeration working solution containing a second induced pluripotent stem cell medium, glycol, DMSO and sucrose, wherein the volume content of glycol in the first refrigeration working solution is same to the volume content of the DMSO in the first refrigeration working solution, the volume content of glycol in the second refrigeration working solution is same to the volume content of the DMSO in the second refrigeration working solution, and the volume content of glycol in the second refrigeration working solution is two times the volume content of the glycol in the first refrigeration working solution. The kit is safe and effective when the kit is used to cryopreserve the induced pluripotent stem cells (iPSCs), the thawing rate of the unfrozen iPSCs is high, and normal karyotype and characteristics can be maintained after culture passage.

The invention relates to a cultivation method of genuine salvia, which comprises the following steps of: establishment of a salvia germplasm resource garden, selection of representative genuine salvia germplasm resources, identification of genuine salvia genome karyotype structure, determination of genuine salvia DNA bar codes, artificial mutation to obtain autotetraploid, sexual hybridization to obtain triploid salvia permanent hybrids, detoxification treatment, tissue culture seedling cloning and proliferation, bottle seedling transplanting, transplanted seedling management, new species comparison and testing, determination of the medicinal material yield and the main drug effective components, and genuine salvia new species determination and species identification. The genuine salvia obtained in the invention is, so far, the only population that has two great natural advantages: the polyploid advantage and the heterosis, and the advantages are giant and irreplaceable. Therefore, the method of the invention is an effective method for cultivating genuine traditional Chinese medicine with strong biotic stress and abiotic stress capability by using traditional Chinese medicine genuine functional genomes and feature genes and based on inherent genes of genuine traditional Chinese medicine.

The invention discloses a production method of triploid pseudobagrus fulvidraco, which belongs to the fish culture technology field. Artificial insemination is performed to the pseudobagrus fulvidraco female parent and male parent which are obtained through artificial induced spawning, after 10 to 15 minutes, triploid pseudobagrus fulvidraco is obtained through induction, and the duration time for induction is 10 to 20 minutes. After the triploid pseudobagrus fulvidraco is obtained through induction, the multiple of the chromosome is measured through the technologies, such as the karyotype analysis of chromosome, the flow cytometer, etc., and the triploid pseudobagrus fulvidraco is identified and sieved for culture. The method restrains the releasing time of the second polar body through time control on the artificial insemination, the induction through the physical or chemical method, and control on the induction duration time, and thereby the inductivity and the survival rate of the triploid pseudobagrus fulvidraco are greatly increased, and according to the experiment statistics, the artificial inductivity of the achieved triploid pseudobagrus fulvidraco is more than 50 percent, and the survival rate is more than 80 percent.

The invention discloses a preparation method of chromosome suitable for karyotype analysis of rubus plants. The chromosome preparation method comprises the following specific steps of: material culturing; pretreatment; fixing; storage; dissolution; dyeing, smearing and tabletting; and microscopic examination and observation. The preparation method disclosed by the invention has the advantages of low cost, simple, convenient and speedy operation process and capability of obtaining an ideal chromosome specimen for karyotype analysis of the rubus plants.

The invention clones insect specific neurotoxin gene BmkIT of East Asian scorpion and insect chitinase gene Chi to an insect cell-converting medium carrier pAcPDIEI to obtain a recombinant plasmid containing divalent anti-insect genes pAcPDIEI-Bmkit-Chi and transfecting the recombinant plasmid and DNA of AcNPV genome together in sfg cells, and obtaining a recombinant rhabdovirus containing divalent anti-insect genes AcNPV-BmKIT-Chi. The cooperation of the insect specific neurotoxin with the chitinase improves the poisoning effect of virus on cotton bollworms and other lepidopteral pests.

The invention discloses a flaking method of petunia chromosome. The method comprises the following steps: getting a petunia young bud at 8:00-10:00 in the morning of a clear day, placing in 0.002mol.L<-1> 8-hydroxyquinoline aqueous solution for pre-treating at low temperature, transferring into a Carnoy's fluid to be fixed, performing low-permeation through 0.075mol.L<-1> potassium chloride, and then water-bathing at 60 DEG.C in 1mol.L<-1> HCl solution for decomposing, washing and then low-permeating through the distilled water, and staining through aceto-carmine staining solution, transferring the young bud onto a glass slide, cutting to get a pistil stigma part, flaking and performing microscopy. The bud size, the pretreatment method, the decomposition and staining method and time and the flaking processes of petunias with different ploidy at optimal sampling time are determined, the obtained chromosome is uniform in dispersion, easy to count and easy for performing karyotype analysis, the operation is simple and convenient, the efficiency is high, and the method has a good application prospect.

A method for expansion of human embryonic stem (hES) cells in a medium including human umbilical cord-derived mesenchymal stem cells (HUCMSCs) as a feeder is provided. The human embryonic stem cells (hES) maintain the features of embryonic stem cells in the medium, such as pluripotency, unlimited undifferentiated proliferation and normal karyotypes. Also provided is a method for non-tumorigenic expansion of the human embryonic stem cells (hES) that is free from forming teratoma.

The invention relates to the technology of antigen protein expression, and particularly relates to a method for preparing a cyprini herpesvirus II antigen coated polyhedrosis based on a baculovirus expression system. According to the method, by designing and recombining bombyx mori nuclear polyhydrosis virus BmNPV-VP3-cyHV-polh, 1-186, 993-1197, 603-783 and 85-186 regional sequences of ORF72, ORF66, ORF81 and ORF82 and the coding sequence of the 1-279 region of a VP3 gene of bombyx mori cytoplasmic polyhedrosis virus structural protein are connected in series to form a fused sequence which is controlled by baculovirus P10 promoter; and the bombyx mori cytoplasmic polyhedrosis protein gene is controlled by a polyhedrin gene promoter of baculovirus. The virus is used for inoculating bombyx mori or bombyx mori culture cells, the recombinant virus expressed cyprini herpesvirus II antigen protein can be coated in bombyx mori cytoplasmic polyhedrosis; the formed polyhedrosis can be purified by simple differential centrifugation; the purified polyhedrosis is cracked under a basic condition, the polyhedrosis protein can be precipitated by centrifuging, and the cyprini herpesvirus II antigen is reserved in supernatant, so that the cyprini herpesvirus II antigen can be quickly and conveniently obtained.

The invention provides a chromosome karyotype image intelligent evaluation method and system. The method relates to the technical field of image processing, and is characterized in that automatic labeling of chromosome image data is carried out through a weak supervised learning model; annotation costs is reduced, labeling efficiency is improved, extension modes such as random projection transformation are adopted for chromosome image data; an optimized residual network model is adopted to build a scoring network model, detail differences of chromosome images are better reserved, the accuracyof the scoring network model is improved, meanwhile, a scoring system is established by adopting a micro-service distributed frame through a cloud platform technology, cross-cloud image analysis is achieved, and the efficiency of the system is improved.

The invention discloses a method for finding a protein interaction with baculovirus PCNA (Proliferating Cell Nuclear Antigen). The method is characterized by comprising the following steps of cloning full-length DNA of an autographa californica nuclear polyhedrosis virus (AcMNPV) PCNA gene by utilizing the means such as molecular cloning, and DNA sequencing, purifying the full-length DNA, coupling the full-length DNA to an affinity column, and fining the interaction protein by adopting the mass spectrography. The method can be used for finding and functionally studying interaction protein of the baculovirus PCNA.