73 results about "Nucleoplasm" patented technology

The invention discloses a method for agrobacterium tumefaciens-mediated genetic transformation of sugarcane. The method comprises the steps: leading agrobacterium tumefaciens liquor carrying plant expression vectors to infect embryogenic callus of the sugarcane; and selecting and proliferating resistant calli to induce the resistant calli to obtain the resistant buds of the sugarcane, wherein matrix attachment region sequences are connected at two sides of a gene expression box of the plant expression vector. According to the method, MARs (metal artifact reduction) sequences are built at two sides of the gene expression box of the plant expression vector, thus greatly improving the exogenous gene transformation efficiency and seedling rate of the sugarcane, and being simple in process flow and low in cost.

The NF-E2-related factor 2 (Nrf2) is a key transcriptional regulator of antioxidant defense and detoxification. To directly monitor stabilization of Nrf2 we fused its Neh2 domain, responsible for the interaction with its nucleocytoplasmic regulator, Keap1, to firefly luciferase (Neh2-luciferase). It is shown herein that Neh2 domain is sufficient for recognition, ubiquitination and proteasomal degradation of Neh2-luciferase fusion protein. The novel Neh2-luc reporter system allows direct monitoring of the adaptive response to redox stress and classification of drugs based on the time-course of reporter activation. The novel reporter was used to screen a library of compounds to identify activators of Nrf2. The most robust and yet non toxic Nrf2 activators found—nordihydroguaiaretic acid, fisetin, and gedunin-induced astrocyte-dependent neuroprotection from oxidative stress via an Nrf2-dependent mechanism.

This invention discloses a method for imaging live nucleus and cytoplasm, which comprises the following steps that: (a) live cells are irradiated by a near-infrared femtosecond laser which is focused by a microscope; (b) the live cells produce two-photon fluorescence and second harmonic under the action of the near-infrared femtosecond laser focused by the microscope; and (c) the two-photon fluorescence and the second harmonic are subjected to microscopic imaging at the same time, the two-photon fluorescence performs microscopic imaging on target molecules which are marked by fluorescence, and the second harmonic performs microscopic imaging on the nucleus. The invention also discloses application of the method for imaging the live nucleus and cytoplasm in monitoring a live nucleus and cytoplasm signal pathway, eliminates photoinduced toxicity and photobleaching, and can observe the live cells for a long time without affecting the activity of the cells and furthest maintain the functional environment of the cells. The method can be applied to the level layer research of cell molecules in a tumor generation mechanism and the research of a pharmacodynamical mechanism of traditional Chinese medicinal herbs.

The invention discloses a whole breast subserial large slice preparation method. The method comprises the following steps of collecting a tumor sample; performing continuous parallel splitting to obtain sample blocks which are split in parallel; performing soaking and fixing in a neutral formaldehyde solution with the concentration of 4% for 24 hours; removing redundant fat tissues by clipping; performing continuous parallel cutting in parallel to an original slice again to form a slice shape; performing continuous fixing for 48 hours by using the neutral formaldehyde solution with the concentration of 4%; performing sequential dehydration by adopting gradient ethanol; performing transparency treatment by using xylene; performing wax dipping in an electric heating constant temperature incubator at 62-65 DEG C to obtain a paraffin pathological sample; and slicing the paraffin pathological sample by adopting a rotary paraffin slicing machine, wherein each sample block can be subjected toconventional pathological staining including HE through continuous slicing. The thickness of a large slice prepared with the method can reach the conventional slice thickness of 4-5 microns; the slice is complete in tissue structure, smooth, clean and free of folds and pollution; no slice stripping phenomenon occurs; the cell nucleoplasm proportion is clear; and a staining result is consistent with a conventional clinical staining result.

The invention belongs to the technical field of rape molecular breeding, and specifically relates to a breeding method of cabbage type rape woad oil cytoplasm male sterile line. The invention also relates to a protoplast fusion technology to recombinate the cytoplasm and nuclear genome of cabbage type rape-woad so as to establish cytoplasmic male sterility of recombinant cytoplasm. The breeding process comprises the following steps: obtaining the leaf meat protoplast of cabbage type rape-woad through separation and extraction, obtaining the protoplast fused somatic cell hybrid of cabbage type rape-woad through a cell fusion method; taking the obtained somatic cell hybrid of cabbage type rape-woad and leaf meat protoplast cell hybrid of cabbage type rape-woad as the female parent, taking cabbage type rape as the male parent, carrying out multi-generation backcross, and obtaining cytoplasmic male sterile line with carpelloid stamen; wherein the mitochondrion gene PCR amplification verifies that mitochondrial genome carries out recombination during the fusion process, thus the nucleoplasm becomes uneven, and the sterile character appears.

The invention discloses a method for transforming a hot-pepper nucleo-cytoplasmic interactive male-sterile line, a homo-maintainer line and a homo-restorer line, which comprises the following steps of: hybridizing by using a hot-pepper selfing line as a male parent and the hot-pepper nucleo-cytoplasmic interactive male-sterile line as a female parent, simultaneously selfing a male parent plant, selecting sterile plants or other inflorescences in 4-50 corresponding male-parent filial single plants and hot-pepper filial generations for paired test crossing when in filial-generation fertility separation, selfing each male-parent single plant, and respectively processing according to the group fertility expression of test crossing generations to select the male-sterile line, the homo-maintainer line and the homo-restorer line. When in filial-generation group fertility separation, the invention not only can be used for transforming the hot-pepper nucleo-cytoplasmic interactive male-sterile line and the homo-maintainer line from the hot-pepper selfing line, but also can be used for breeding the homo-restorer line, expands the selecting space of the restored male parents of other homologous sterile lines and can be used for screening more excellent three-line combined varieties. A hot-pepper hybrid is produced by adopting a male-sterile three-line technology, all the complex pollination procedures are simplified, and the economic benefit is quite remarkable.

The invention relates to a cell sorting method for an affine propagation clustering, which comprises the following steps: (1) selecting a circularity parameter C and a rectangularity parameter R of a cell image, designing a sample coordinate X sample=lambda C+(1-lambda) R, selecting an area parameter Area of the cell image as another sample coordinate Y sample and selecting a nuclear-cytoplasmic ratio parameter prop of the cell image as another sample coordinate Z sample, wherein lambda represents a prior input value; (2) using euclidean distance of the three-dimensional sample coordinates as sample distance, wherein a diagonal value of an S matrix of the affine propagation clustering is an average value of distances among the samples; (3) under the initial condition, setting an membership grade matrix A(i, k)=0, updating a matrix R and updating a matrix A; and (4) stopping after the number of iteration times is set and obtaining different types of cells from a sorting result. The invention provides the cell sorting method for the affine propagation clustering, which is suitable to process mass data, has excellent real-time property and can effectively carry out cell sorting.

The present invention relates to plant breeding technology, and is especially anther cultivation technological method of breeding unclear sterile dual purpose line and unclear substance sterile recovery line of capsicum. Extracorporeal anther cultivation technology is used in selective breeding of unclear sterile dual purpose line and unclear substance sterile recovery line of capsicum. That is, the dual purpose line is obtained through test cross of the fertile plant and sterile plant in contemporary anther cultivation offspring of dual purpose line hybrid, F1 selfing, test cross of similar fertile plant and sterile plant in the offspring lines and continuous two generation selection; and the recovery line is obtained through the test cross of the fertile plant and sterile plant in the contemporary anther cultivation offspring of three line hybrid with available sterile line, selectively breeding the F1 with fertility recovering rate of 100 % and stable fertility to obtain fertile plant as the corresponding recovering plant and final selfing.

The invention discloses a breeding method of a polima genic-cytoplasmic male sterile line. The method mainly comprises the steps of obtaining a cytoplasmic male sterile plant by hybridizing, backcrossing, multiple crossing and an anther culture method; performing hybridizing, multi-generation backcrossing or multi-generation sister-crossing with strains with restoring gene, preparing a target sterile line and a target maintainer line; taking the target maintainer line as a recurrent parent, performing hybridizing and backcrossing with strains with fertile cytoplasm; replacing the sterile cytoplasm of the target maintainer line with fertile cytoplasm; after selfing of the cytoplasm-replaced target maintainer, performing test-crossing with the target sterile line; and identifying sterile maintaining ability of the maintainer plant and breeding the sterile line. The method can effectively remove minor restoring gene, improve outcrossing ability of the sterile line, improve breeding efficiency of the male sterile line, obtain excellent sterile line, breed full maintenance line from the sterile cytoplasmic resource which is difficult to abort thoroughly, expand utilization range of the polima genic-cytoplasmic male sterile resource and enlarge restoring spectrum of the male sterile line.

A selective culture method for the male sterility line of paddy rice features that the sterile cytoplasm of nucleoplasmic interaction type male sterility line of paddy rice is used to realize the hybridization between maintenance line and other excellent variethes, and the cultured said nucleoplasmic interaction type male sterility line as female parent and the male sterility recovery line as male parent are hybridized to obtain new combination. The resultant new combination has the advantages of high adaptability, stress resistance and disease and past resistance, and improved quality and yield of rice.

A reproductive and seed culturing method for the karyoplasmic interaction type incomplete male sterility line of rice includes such steps as choosing male parent and female parent (Karyoplasmic interaction type incomplete male sterility line), sowing, irrigating with cold water when the ambient temp is higher than the critical temp of fertility transform, hybridizing, growing and harvesting.

The invention provides a method for improving compatibility and development rate of interracial nuclear transfer embryo nucleoplasm. The method includes the steps: (a) a cell nucleus of a nuclear donor cell is moved into a perivitelline space of an enucleated oocyte of a non-human mammal and integrated by means of electric stimulation, so that a reconstructed ovum is formed, (b) a human oocyte is moved into the reconstructed ovum in the step (a), and (c) the reconstructed ovum in the step (b) is electrically stimulated so as to form a reconstructed ovum composed of the cell nucleus of a human cell, the enucleated oocyte of the non-human mammal and the human oocyte. The embryo development rate can be increased remarkably by the method.

The present invention belongs to the field of bioengineering technology, and is especially biological consubstantial cell nucleus grafting and cloning technology. By means of biological cloning technology, the present invention takes consubstantial nucleus donor cell and denucleated egg mother cell with clear genetic background for consubstantial cell nucleus grafting to prepare cloned animal. The present invention has relatively high matching between the donor nucleus DNA and acceptor cytoplasm and the electronic compatibility inside nuclear substance, and can improve the development of cloned embryo and raise cloning efficiency. The said technology may be used in cloning animal to obtain high quality livestock.

A method of creating a model of a biological cell. The method features providing a circular base with indentations disposed therein, forming a cell membrane and at least one organelle from a mixture formed by mixing all-purpose flour and water. Organelles may include a nuclear membrane, a cell wall, mitochondria, a smooth endoplasmic reticulum (ER), a rough ER, a vacuole, a golgi apparatus, a nucleolus, a nucleoplasm, chromatin, a ribosome, DNA, RNA, transcription machinery, a histone, and a microtubule. The outside edge of the base is lined with the cell membrane. The organelles are inserted into indentations or atop the base. The organelles can be secured with glue.

A method for preparing the seeds of hybrid crops by using the minor restoring gene of nuclear-cytoplasm interactive male sterility line includes such steps as choosing the nuclear-cytoplasm interactive male sterility line A, its maintenance line B and its recovery line R, preparing the subisopolar gene line B' or B' of said B and with the minor restoring gene, discriminating the fully male sterility A/B' F1 or A/B'//B' F1, and using A/B'//R or A/B'//B'///R to produce the seeds of hybrid crops.

A method for culturing a novel sterile line of paddy rice is characterized by that the cytoplasm of male sterility line of non-flutinous rice is used as the doner of the cytoplasm for novel sterile line and the photosensitive karyosterile line of non-glutinous rice containing male sterile maintainer gene is used as the maintainer line of novel sterile line. Its advantage is full male sterility in either longer or shorter day condition.

The invention discloses a method for breeding onion male sterile line and maintainer line by utilizing molecular markers. A PCR (polymerase chain reaction) experiment is performed to detect single onion ball nucleoplasm genotype by utilizing specific primers (disclosed as SEQ ID NO.1-6) to screen a sterile strain and a maintainer strain onion ball, thereby implementing two-line matching. A plurality of sets of primers are designed according to Ms allele flanking sequences (F1890 and S1887) and onion cytoplasmic male sterility specific sequence mt472; and scientific experiments are performed to develop a molecular marker detection system and detection kit based on multiplex PCR. The detection system can distinguish 6 karyoplasm fertility types of onions through one PCR experiment; and the detection system is utilized to directly breed the sterile line and maintainer line from the open pollination colony, thereby implementing one-step matching of two lines. The development of the multiplex PCR detection system has important meanings in accurate and quick identification of onion nucleo-cytoplasmic interreaction male sterility karyoplasm genotype.

The invention discloses a culture method of dental pulp mesenchymal stem cells. The culture method comprises the following steps: preparing a dental pulp sample, separating and culturing the dental pulp mesenchymal stem cells, carrying out colonized culture on separated dental pulp mesenchymal stem cells, detecting the dental pulp mesenchymal stem cells, and detecting and staining the dental pulpmesenchymal stem cells. The culture method of the dental pulp mesenchymal stem cells disclosed by the invention has the beneficial effects that 1, the cells cultured by being cloned and separated areclosely arranged, so that central cells are unclear in boundaries, round or irregular and cell nodules are formed, peripheral cells are polygonal or short fusiform, less in cytoplasm, large in proportion of nucleoplasm and obvious in nucleoli and some cells are fibrillar; 2, the problem of serious immunological rejection is not generated; 3, the dental pulp mesenchymal stem cells are convenient intaking materials and safe and small in cross-infection risk, thereby being capable of being used for repairing defective teeth and tooth regeneration; and 4, the dental pulp mesenchymal stem cells can promote skin wounds to heal and regenerate, delay aging or cure blindness, and further can be used for treating treat heart diseases, rheumatoid arthritis, burns, stroke or cartilage damage and thelike and is wide in use.

The invention provides a method suitable for in-situ hybridization of a cherax quadricariratus gonadal tissue mRNA paraffin section, and belongs to the technical field of in-situ hybridization. The method comprises the steps of gonadal tissue embedding, paraffin sectioning, in-situ hybridization, and photographing and recording. The sequence of a probe used in in-situ hybridization is as shown inSEQ ID NO.3. The probe has clear specificity on tissue positioning of a dsx gene, so that the display effect on the mRNA level of the dsx gene is realized. The method for mRNA in-situ hybridization ofthe paraffin section is established in cherax quadricariratus to research expression modes of related genes in the gonad, and the expression location of the sex-related genes of the cherax quadricariratus in the gonad can be clearly described. The method avoids tissue shrinkage and cell deformation while the dewaxing effect of the paraffin section is good, so that the tissue is still kept in theoriginal position, the tissue morphology is kept complete, the enzymolysis effect can be improved, cell coloring is good, nucleoplasm is clear, and the in-situ hybridization accuracy is finally improved.

The invention belongs to the technical field of agricultural seed breeding and discloses a method for rapidly breeding nucleo-cytoplasmic interaction sterile lines. The method comprises the following steps of: utilizing the existing male sterile lines and a plurality of homozygous and excellent selfing line male parent materials to carry out paired testcross; carrying out simultaneous selfing on male parents, and analyzing offspring fertility; selecting 12 sterile plants in a fertility-separation testcross combination for one-to-one backcross with male-parent selfing offspring; and rapidly transbreeding out male sterile lines and maintainer lines in a third-generation backcross combination; and the method comprises the detailed steps of: screening a first generation, screening a second generation and screening a third generation. The method disclosed by the invention has the advantages that excellent selfing lines can be rapidly transbreeded into the male sterile lines, the maintainer lines are screened out, and the sterile degree reaches 100%; 12 plants are selected in the male-parent selfing offspring to carry out testcross with 12 sterile plants in F1 generation for three times repeatedly, so that the probability of obtaining the sterile lines and the maintainer lines is increased.

The invention discloses a preparation and in-vitro culture method of interspecies-cloned yak embryos. According to the method, yak-cattle interspecies-cloned embryos are constructed by using yak mammary glandular cells as donor cells and cattle or milch cow enucleated oocytes as receptor cytoplasts by a somatic nucleus transplantation technique; and an electrical activation-chemical activation-reelectrical activation mode and a self-made sequential culture system are utilized to acquire the high-efficiency high-activity yak-cattle interspecies-cloned embryos. The method disclosed by the invention can increase the yak breeding speed, shorten the yak breeding cycle, lower the yak production cost, enhance the breeding efficiency, reduce the damage level of the grassland to some extent, and effectively protect the species resource of the yak. Besides, the invention has important meanings for promoting research of interspecies-cloned yak, transgenic cloned yak embryos and nucleo-cytoplasmic interaction.