1088 results about "Zygote" patented technology

The invention relates to a method of constructing an Ifit3-eKO1 gene knockout mouse animal model and belongs to the biotechnical field. The method comprises the following steps: S1, determining specific target sites sgRNA1 and sgRNA2 of a to-be-knocked out gene of an Ifit3-eKO1 mouse, and performing in vitro transcription with Cas9 nuclease to mRNA; and S2, micro-injecting active sgRNA and Cas9RNA into an oosperm of the mouse to obtain the Ifit3-eKO1 gene knockout mouse. The method has the advantages that by using the CRISPR/Cas9 gene knockout technology, the Ifit3-eKO1 gene knockout mouse animal model is constructed for the first time, thereby providing a convenient, reliable and economical animal model for researching action of Ifit3 in tumorigenesis and development.

The present invention is artificial breeding method and incubator for Shatang carp. The artificial breeding method includes the steps of parent fish breeding, spawning and fertilization, hatching and carp fry culturing. After intensified parent fish breeding, temperature control, flow water stimulation and other method are taken to promote the synchronous spawning and fertilization of parent fishes, the fertilized eggs on tiles are sterilized and hatched in a special hatching device, and the hatched carp fries are cultured in a special cement pond. The artificial hatching device is one rectangular hatching tank with shield, ventilating water circulating pump, water level and water flow controlling mechanism. The technology of the present invention is suitable for large scale production of Shatang carp fry.

The invention relates to a method of industrial and artificial breeding of Epinephelus, which is applicable to the breeding of Epinephelus malabaricus and E.coioides. The method comprises the following steps: (1) treating a nursery pond and water; (2) preparing before breeding; (3) putting larval fishes of Epinephelus or germ cells; (4) controlling the quality of the breeding water body; and (5) feeding baits. The method is applied to conduct the industrial and artificial breeding, so that the water quality of the breeding is stable, the output for the unit water body breeding is high, the bred sizes of advanced fries are uniform, no antibacterial drugs are used, no special breeding is conducted to unicellular alga, no bottom-attaching happens in the process of breeding, and the operation is simple.

The invention discloses a mimic ecological propagation method for breeding parent fish of American hilsa herring, comprising the following steps: establishing a mimic ecological breeding system suitable for natural growth and maturity of the sexual gland of hilsa herring by adopting a circulation water breeding technique and an automation control technique of water environmental factor; regulatingthe mimic natural seasonal rhythmicity for the major physical factors, water chemical factors, and the like of the breeding environment, including factors such as salinity, water temperature, water flow and period of lighting daily variation while fortification; injecting hormone to the parent fish before the spawning season to promote maturity of the sexual gland, and causing the hilsa herring to spawn naturally through enhance water flow stimulation; arranging a spawn collecting box at a circulation water outlet to collect the germ cells and then placing the germ cells in a semi-floating germ cell separation incubator for incubation. The invention can remarkably improve the spawning efficiency of the parent fish of hilsa herring and the incubation rate of the germ cell, and provide thetechnical support for the industrialization of artificial propagation technique for breeding hilsa herring.

The invention discloses an animal sex control method based on Rbmy gene editing and application. The method controls animal sex by using CRISPR/Cas9 system to specifically cut Rbmy gene on Y chromosome. Rbmy gene encodes a germ cell specific nucleoprotein, this protein is an important factor related to spermatogenesis, and by cutting Rbmy gene, it is possible to deactivate Rbmy gene, thus Y sperm or fertilized ovum with Y sperm is deactivated and cannot develop into a normal embryo, birth ratio of female animals is finally increased and sex control is achieved. Therefore, sex ratio of animal offspring can be controlled through the method so that the ratio of female animals with better economic value is increased and production efficiency and economic effect are greatly improved.

Synchronous induced spawning for parent fish better solves the problem of sexual maturity through synchronous development of female and male parent fish. The invention discloses an artificial propagation method for sturgeon, which comprises the steps of selection of parent fish, parent fish rearing, induced spawning, artificial insemination, zygote hatching, artificial selection of fish fry, and fish fry rearing. In the artificial propagation method, by artificially reducing water temperature and ambient environment temperature, micro water flow is formed to simulate the ecological environment for the reproduction of sturgeon, the simulated ecological environment is similar to the conditions necessary for the reproduction of wild sturgeon in the natural world, and the artificial low temperature factor lasts for a long time, so as to always guarantee the low temperature environment for the maturity and awaiting delivery of parent fish of the sturgeon, and meet other condition requirements, so that the parent fish can rut and spawn all year round to finally achieve the factory production of the sturgeon fry; meanwhile, in the artificial propagation method, female parent fish and male parent fish of the sturgeon are separately put into an artificially simulated ecological environment for inducing oestrus, and then the degree of sexual maturity of the parent fish is identified to select the parent fish with basically developed sexual maturity.

A method for culturing the minnow of roughskin sculpin includes such steps as capturing the parent fish in near sea in early spring, temporarily culturing in indoor pool, using flowing water to stimulate the parents for estrus, natural mating, ovipositing, collecting fertilized ova, natural incubating in natural seawater in pool, regulating illuminance for correcting the strong phototaxis, applying nutritive forage, improving water quality, and post culturing.

The invention relates to a technical method of triploid seed production of rainbow trout. The technical method comprises the following steps: in proper time after sperm-egg binding, carrying out thermal (shock) treatment on oosperm in low-temperature environment in a certain time interval, then restoring the original low-temperature environment and carrying out cooling water incubation; leading more than 80% of oosperms which need to be incubated into diploid oosperm to generate triploid oosperm after thermal shock treatment; and raising the triploid oosperm into triploid offspring seed (larval fish). The implementation of the technical proposal stops the discharging of a secondary polar body of the oosperm, induces the oosperm to produce another type of oosperm which is fused by the egg, the sperm nucleus and the secondary polar body with respective monoploid chromosome and contains triploid chromosome, namely triploid oosperm, and realize industrialization of triploid seed production. Each thermal inducing incubator can produce more than 40,000 of triploid oosperms for one time, and each reproduction period (October to February) can produce more than 2,000,000 oosperms. The inductivity of the triploid is more than 80%.

The invention discloses a CRISPR/Cpf1 gene editing method applicable to a marine crustacean mitochondrion genome. The method includes the steps: introducing MLS signals positioned by a marine crustacean mitochondrion into a CRISPR/Cpf1 expression plasmid; designing gRNA; micro-injecting fertilized eggs; culturing the fertilized eggs in an in-vitro manner; detecting editing effect to complete editing of the marine crustacean mitochondrion genome. By the method, the marine crustacean mitochondrion genome can be directionally edited, and the method has significant value to a condition that artificial proliferation releasing markers are introduced into the marine crustacean mitochondrion genome.

The invention provides a method for specifically knocking off a microRNA-8086 gene by using CRISPR-Cas9 (Clustered Regularly Interspaced Short Palindromic Repeats). The method comprises the following steps: (1) establishing a Cas9 expression vector; (2) establishing a sgRNA expression vector; and (3) mixing Cas9 and sgRNA- so as to obtain a mixed liquid, and injecting the mixed liquid into zygotes of mammals. Therefore, the purpose of fixed point knock-off of the microRNA-8086 gene is achieved.

The invention relates to the technical field of the breeding of shellfish seedlings, in particular to an artificial breeding method for Mactra chinensis. Through the combination of artificial ripening and ecological ripening of parents, the development of sex glands is regularly detected to determine the mature period of the sex glands of the parents; methods such as shade drying, water flowing and so on, are adopted to stimulate the spawning and spermiation of the parents to obtain fertilized eggs, and D larvae are obtained through the embryonic development; and on this basis, larva breeding, larva metamorphosis and juvenile shellfish cultivation are performed. A large amount of Mactra chinensis seedlings can be obtained through the links. The method breaks through an artificial breeding technology of the Mactra chinensis, solves the problem of the breeding of the Mactra chinensis seedlings that fails to be broken through for a long time, and lays a solid foundation for the development of cultivation and proliferation of native species, the resource recovery of shallow seas and tidal-flat areas, and the maintenance of biological diversity. The method has the characteristics of simple and convenient operation and high practicality.

The present invention relates to an artificial breeding and pisciculture method of murrel, including the following steps: selecting piscicultural pond, breed selection of parent fish and cultivating,artifically-hastening parturition, spawning and fertilization, fish roe incubation and fish fry cultivation. In the course of spawning and fertilization, after the fertilized eggs are incubated and the fish fries are obtained, the parent fishes can be taken out. The health mature parent fish can be used as seed fish, and the ratio of male and famel is 1:1, and its propagation coefficient and percentage of success are high.

The present invention relates to microscopic tissue section technology, and establishes one effective tissue section method for the fertilized egg of sea water left eyed founder and right-eyed founder based on the features of the fertilized egg of sea water left eyed founder and right-eyed founder. The fertilized egg is polylecithal pelagic egg with high egg membrane toughness and narrow perivitelline space. The tissue section method includes the following steps: 1. fixing the polylecithal pelagic egg with Bouiní»s fixing fluid and optimizing treatment period; 2. pricking the egg membrane with dissecting needle in the vegetative pole direction to peel off the egg membrane; and 3. directly embedded to fix embryo with hot agar. The method is simple and effective, and may be also applied in the tissue section of other sea water fish egg.

The invention relates to a method for comprehensively regulating and controlling gonad mature ovulation of cheilinus undulates, which comprises the following steps: preferably selecting, breeding and feeding parent fishes; regulating and controlling water temperature, illumination and water flow to enable the parent fishes to grow; and enhancing the nutrition to promote gonad mature ovulation. The technical method is used for screening artificial ripening effective temperature, intensity of illumination and water flow velocity factor data, screening varieties and doses of artificial ripening effective nutrition enhancers, screening the time suitable for gametogenesis and fertilization of cheilinus undulates, and acquiring high-quality fertilized eggs of cheilinus undulates. The fertilization rate of the eggs of the cheilinus undulates acquired by the method is 20-86%, and the hatching rate is 30-78%. The technical method for comprehensively regulating and controlling gonad mature ovulation of cheilinus undulates, which is provided by the invention, has the characteristics of advancement, high efficiency, practicality and reliability and comprehensively regulates and controls temperature, illumination, water flow and nutrition enhancement in the breeding process of seedlings of cheilinus undulates. The invention has great application value and wide popularization and application prospects for protecting endangered species.

The invention provides a method for constructing KARS-gene-mutated mouse models and application of the KARS-gene-mutated mouse models. The method for constructing the KARS-gene-mutated mouse models comprises the following steps: (1) carrying out target point construction: looking for a gene corresponding to nervous system diseases, and carrying out target point design; (2) designing Guide-RNA/Cas9mRNA, preparing plasmid, carrying out transcription, and then, injecting a transcription product into cytoplasm of a fertilized egg in a microinjection manner; (3) carrying out Founder mouse identification; (4) carrying out F1 mouse identification. According to the method for constructing the mouse models with the nervous system diseases and the application of the mouse models, provided by the invention, site-directed mutagenesis aiming at a KARS gene is carried out, so that a disease research on the model mice can be carried out under the condition of being closer to the occurrence and development of actual diseases, and a research result has a higher reference value.

The invention relates to a method for in vitro incubation of fertilized eggs of macrobrachium nipponense, which is characterized in that the method comprises the steps of selection of parent shrimps, mating and oviposition, treatment of incubation water, egg stripping, egg washing, incubation, daily management and the like. The method for the in vitro incubation of the fertilized eggs of the macrobrachium nipponense solves the problem of biological characteristics that the fertilized eggs depend on parent bodies to be incubated, lays a foundation for genetic operations of the macrobrachium nipponense such as gene transfer, nuclear transplantation and the like, fills the research gap of the field, and has important practical application value.

Instead of immersing human reproductive cells in a single culture medium throughout the various procedures used in IVF, a process is provided by which the reproductive cells may be moved through a sequence of distinct culture media as the various IVF procedures are carried out. In one implementation, the culture media specifically formulated to provide a physical environment similar to that found within the female reproductive tract and conducive to growth and development of human reproductive cells during the various stages of the IVF process. In this regard, specifically formulated culture media can be applied to support the reproductive cells in one or more of the following procedures: oocyte retrieval and handling; oocyte maturation; ordinary fertilization; oocyte, zygote and embryo examination and biopsy; embryonic development to the eight-cell stage; embryonic development to the blastocyst stage; embryo transfer; and cryopreservation.

The invention relates to a method for cultivating glyptosternum maculatum fries and fingerlings through artificial feed. In an indoor cement pool, clean water resources from rivers are utilized, micro water flow is controlled through manpower, manual mixed feed is provided for feeding, and fries hatched from germ cells are cultivated to fingerlings 3cm in length. The rate of survival of the glyptosternum maculatum fries which live for 2-3 days to be fingerlings which are 3cm long is more than 80%.

The invention relates to a method for cultivating inkfish fries and belongs to the technical field of aquatic product culture. The method is characterized in that the inkfish fries are cultivated by utilizing a shrimp pond and managed according to the growth characteristics of inkfish fries. The method comprises the following steps: preliminary preparation, parent spawning, zygote acquisition, hatching, biological bait cultivation, stage-I cultivation of young inkfish, stage-II cultivation of young inkfish, stage-I cultivation of fries, stage-II cultivation of fries, domestication by food and the like. By adopting the method, the palatability of the young inkfish can be maximally satisfied, the survival rate is increased, the economic benefit is increased, and the method is suitable for being popularized and use.

The invention discloses a full-artificial breeding method for megalonibea fusca. The method comprises the following steps of: getting parent fish seeds from natural sea areas; carrying out reinforced breeding on parent fish; carrying out artificial spawning on the parent fish; carrying out hatching on fertile eggs; carrying out indoor artificial cultivation on juvenile fish; carrying out indoor nutrition strengthening cultivation on the juvenile fish in an alevin stage; carrying out ecological breeding on the alevin which are moved into an earthen pond after the alevin stage; and breeding thealevin into large-size seedlings, and the like. The full-artificial breeding method provided by the invention is a mode applicable to the large-scale production of megalonibea fusca fries. After carrying out three-year cultivation on first-generation juvenile fish which is successfully artificially bred, sexually-matured first-filial-generation parent fish is cultivated, and then second-generation juvenile fish is bred from the first-filial-generation parent fish, thereby achieving the full-artificial breeding. The full-artificial breeding method breaks through the bottleneck that because themegalonibea fusca belongs to endangered fish species, parent fish sources are extremely difficult, and realizes the large-scale artificial breeding and culturing industrialization of the megalonibea fusca.

The invention discloses a method for establishing a full-female pelteobagrus fulvidraco breeding population, which comprises the following steps of: 1) manually hastening parturition and manually inseminating; 2) reversing sex; 3) identifying inheritance sex of the sex reversal pelteobagrus fulvidraco; 4) expanding the population; and 5) establishing the full-female pelteobagrus fulvidraco breeding population, so as to finally establish the full-female pelteobagrus fulvidraco breeding population. According to the invention, an integrating technology of hormone sex reversal combined with pelteobagrus fulvidraco chromosome peculiar molecular markers is adopted, so that XX physiological milter can be confirmed without adopting a test mating method and the breeding time is shortened; compared with the test mating method, one generation can be shortened; the gynogenesis step is omitted, so that the damage to the germ cell genetic material caused by manual induction gynogenesis is reduced; the yield of the XX raun is increased; and the economic and social benefits are obvious.

The invention discloses establishment of a carrier based on a fish CRISPR/Cas9 system by using a gene knockout method ad an establishing method of the carrier, and relates to a carrier based on the fish CRISPR/Cas9 system and an establishing method of the carrier, so as to solve the problems that in a conventional transgenosis method the position that transgenosis is inserted into genome is hard to confirm and potential safety hazard can be generated when external DNA is led into cells and is randomly integrated by using a microscopic injection method, an electroporation method, a sperm carrier method and the like, and the problems that a conventional gene knockout method is complex in process, high in requirement on techniques, high in expense and long in experiment period, and the success rate is limited by multiple factors. By co-ejecting two non-integrated knockout carriers, namely, pX330-MSTN a and pX330-MSTN b, into zygotes of fishes in the 1-2 cell stages, high-efficiency and rapid gene knockout can be achieved.