112 results about "Zygote formation" patented technology

The invention discloses a method for producing transgenic mice through a homologous recombination technology. The method comprises the following steps: replacing mouse NGF genes on mouse chromosomes by human NGF genes through a Cas-9/CRISPR gene knock-in technology, and obtaining the genes with homozygous human NGF genes through breeding and knocking the genes in mice, thus obtaining filial generation mice with salivary glands capable of secreting human NGF. Compared with the conventional targeting technology utilizing positive and negative double screening homologous recombination genes, the method disclosed by the invention is simple to operate, capable of being realized only by transfecting mouse embryonic stem cells by three plasmid DNAs or carrying out pronucleus injection on the zygotes of mice, and high in success rate which is up to 2-5% and remarkably higher than the mouse embryonic stem cell positive rate of 0.1% of the common gene targeting technology.

The invention relates to an ecological breeding method for sea horses, which is characterized by comprising the following steps: putting sea water in which zygotes of fishes, shrimps and shellfishes are filtered into a cement pool, ensuring that the height of the sea water is 0.2 to 0.4 meter lower than that of the cement pool, and keeping the temperature of the sea water between 15 and 32 DEG C and the salinity of the sea water between 1 to 3 percent; putting kelps into the cement pool, and keeping the fresh weight density of the kelps in the cement pool between 0.5 and 2 kg/m; inoculating organic fertilizer to the cement pool to breed bioactive baits; putting the sea horses to the cement pool; and during breeding, keeping the density of the living biological baits in the cement pool. Compared with the prior art, by adopting the method for breeding the sea horses, little bait even no bait is fed, so the cost is low and the pollution is little; and the produced sea horses have high survival rate, large individual and good economic benefit; and the method can be used for supplementing the resources of coastal sea horses, and broadens the path of marine organism resource protection.

Provided is a method for the production of transgenic animals, especially pigs, by the use of nuclear transfer from genetically modified or other embryonic stem cells to either enucleated oocytes which were matured in vivo or in vitro and activated or to enucleated zygotes.

The invention relates to a method for crossing and breeding red Po and john dory. Wherein, it is characterized in that: in winter and spring, selecting adult and healthy wide ones as parent; breeding the parent; at later may and middle June, ejecting odinagogues HCG and LRH-A2 into parents; and artificially seminating while the ratio between female and male is 1:3, and cultivating the zygote in cultivation cylinder or channel. The invention has low cost and easy transportation.

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 provides an artificial propagation method for yellowfin groupers. The artificial propagation method comprises the steps that (1) parents are selected; (2) a parent breeding pond is separated into a deep water area and a shallow water area with water flow communicated, wherein the water depth of the deep water area ranges from 4 m to 5.5 m, and the water depth of the shallow water area ranges from 3 m to 3.5 m; (3) parent fish breeding management is carried out, wherein a, in first 3 weeks during breeding, LED green light and yellow light alternately irradiate the water surface ofthe pond, and after the parent fishes are bred for 3 weeks or longer, the water surface of the pond is irradiated by the LED green light; b, feed is prepared from 3-5 parts of grouper compound feed and 1-2 parts of small fish and shrimp and artificial pellet feed, the parent fishes are fed with the first feed in the first 3 weeks during breeding, and the parent fishes are fed with the second feedafter being bred for 3 weeks or longer; c, the water temperature of the pond ranges from 21 DEG C to 26 DEG C, and water dissolved oxygen is larger than 4.8 mg/L; (4) breeding is enhanced before propagation is carried out; (5) artificial propagation is carried out; (6) zygotes are collected and hatched. According to the artificial propagation method for the yellowfin groupers, the adaptive capacity to environment of the parent fishes is enhanced, the activities of fish shoals are effectively promoted, the feeding rate is effectively increased, the fertility is high, and large-scale artificialbreeding of the yellowfin groupers can be effectively achieved.

An immune fluorescent microscopic observing method of cytula micro-pipe skeleton from bothidae flasfish includes applying fixture solution to fix sample and utilizing dissection needle to separate germ disc from embryon, making hole on sample and using sodium borohydride to eliminate aldehyde group influence on experimental result, carrying out indirect immune fluorescent staining on sample then using fluorescence microscope to observe sample.

The invention relates to a method for breeding sole, which comprises: (1) cultivating parents that 1, adjusting water temperature that naturally decreasing 6-7Deg. C, and increasing to 12Deg. C; 2, adjusting light with lamp; 3, adjusting bait that uses shrimp, fish, etc, with vitamin and lecithin; 4, daily management that clearing and exchanging water; 5, using HCG and LHRH to accelerating growth; (2), picking up semen and eggs, and cultivating zygote, while the zygote is obtained by dry semination, and using sinke zygote and semi-sink zygote to cultivate the baby fish; (3), breeding seed that cultivating 3-5cm seeds in small and large pools, while the total survival rate can reach 28.0-30.0%, and the total survival rate at 8-9cm can reach 21%. The invention can improve the survival rate of seeds.

The invention relates to a feminization method for ZZ zygocyte of Oria tilapia of which the male parent is ZZ and the female parent is WZ, belonging to the field of tilapia cultivation. The technical proposal is as follows: before the gonad of the Oria tilapia starts differentiation, feeding the Oria tilapia with tonic that contains estrogen hormone for more than 15 days, and then using the ordinary way of feeding and management to get ZZ type physiologically female fish, wherein every gramme of tonic contains 100 to 200 microgrammes of estrogen hormone, and the temperature for feeding with the tonic is 25 to 30 DEG C. The invention can change the hereditarily male fish into physiologically female fish, such that it sets up a novel breeding system of ZZ type all male Oria tilapia having significant meaning in the expanding and breeding of all male Oria tilapia.

The invention discloses a method for collecting and separating medaka zygotes, and relates to the technology of fish zygote separation. In order to solve the technical problems, the invention provides the method for collecting and separating the medaka zygotes for preventing egg membranes from being punctured. The invention adopts the technical scheme that the method adopts a soft small brail, disposable engineering plastic inoculation cups, a large-mouth suction tube and culture dishes. The method is characterized by comprising the following steps of: fishing female fishes of which bellies are hung with eggs within 1 to 6 hours after laying eggs to the water surface by using the soft small brail, taking down the egg blocks by using the disposable inoculation cup, and transferring the taken-down egg blocks to the glass culture dish filled with Ringer's culture solution to perform separation; and mutually stretching the inoculation cups held in the left and right hands to break the egg silks and separate the zygotes, sucking the separated single eggs to the other culture dish filled with the Ringer's culture solution by using the large-mouth suction tube to perform hatch, and controlling the hatching temperature to be between 25 and 26 DEG C. The method is used for embryo observation and experiments.

The invention discloses an in-vitro hatching method for freshwater lobster zygote. The technical scheme comprises the steps of: putting oogenesis parent lobsters fished in batch into a methylene blue solution to be soaked for 5-15 minutes; lightly stripping zygotes from parent lobster appendages by using sterile tweezers, and putting into a zygote hatcher; and hatching into post larvae after 16-26 days, wherein the hatching rate of the zygote can achieve more than 95%. The method has the advantages of large zygote amount, little occupation land for zygote in-vitro hatching, little water using amount, labors and materials saving, no need of continuous breeding of the oogenesis lobster, and reduction of feeding management cost of the oogenesis lobster. The embryo can survive, and quickly hatched into larvae by adopting the in-vitro hatching method; the size of the larvae is uniform, and the commercial value is high; the mutual killing phenomenon among the larvae can be effectively prevented, and the survival rate of larvae is enhanced; and the in-vitro hatching method is suitable for the in-vitro hatching of scaled freshwater lobster zygotes.

The invention belongs to the field of fish breeding, and mainly relates to a device for incubating zygotes of fishes. The device comprises an incubating pond. A first water inlet is formed in the bottom of the incubating pond, a second water inlet is formed in a position above the incubating pond, and artificial fish nests for collecting roes are arranged in the incubating pond. A shower head for spraying water into the incubating pond is connected to the second water inlet. The device has the advantages that water is supplied into the incubating pond from bottom to top by the first water inlet in the bottom of the incubating pond, so that water in the incubating pond can be always in a flowing state, sufficient oxygen in the water can be guaranteed, oxygen deficient conditions of the roes due to the fact that the roes cohere with one another can be prevented, and the roes can be assuredly normally incubated; the water can be sprayed on a water surface of the incubating pond from top to bottom by the second water inlet, impact force of the water can be relieved by a shower head structure under the condition that requirements on water changing quantities are met and water flow is guaranteed, and accordingly the zygotes and/or fish fry just out of membranes can be prevented from being burst open by the water flow.

The invention provides a boot method, a boot device and an Android intelligent device. The method comprises the following steps of starting ActivityManagerService in a SystemServer process created by a Zygote process in a boot process; sending custom BOOT-P0 broadcast for starting a system service or a system application with the first priority in a FinishBooting function or other processes by the ActivityManagerService; after the custom BOOT-P0 broadcast is sent for the preset time, sending custom BOOT-P1 broadcast to start a system service or an application whose boot priority is next only to the system service or application started by the BOOT-P0 broadcast; and after the custom BOOT-P1 broadcast is sent for the preset time, sending original ACTION-BOOT-COMPLETED broadcast for starting a third-party application and/or a system service with third priority. The custom BOOT-P broadcast can be increased according to the number of the system services which need to be started, so that the applications or the services to be started are started in a boot process according to the order of the priority, and thus the system lag caused by CPU occupation due to centralized startup of many applications or services at the same moment is avoided, the system burden is relieved and the smoothness of system running is improved.

The invention discloses a method for collecting seedlings from scytosiphon filaments. The heterothallic characteristics of scytosiphon gametes are used for acquiring filaments from discoid bodies formed by parthenogenesis (etheogenesis) of the gametes obtained from individual mature algae, the method for collecting seedlings from filaments is used for chopping and spreading the filaments on an attaching base, and filament cells on the attaching base can directly germinate and form monoploid scytosiphon seedlings. The filaments of propagation culture can be preserved for a long time, the seedling collecting time and the seedling collecting quantity can be determined according to production requirements every year, the culture phases of zygote collection, discoid sporophyte growth, sporangium maturity and the like in the prior art are omitted, and the indoor culture time can be saved by more than 5 months. The invention basically overcomes the defects of season limitation of scytosiphon collection, zygote insemination and alga propagation, long indoor culture time, low success rate of culture of seedlings and the like, reduces the cost of culture of seedlings, greatly improves the success rate of culture of seedlings, provides a large number of seedlings for scytosiphon production, and industrializes the scytosiphon planting industry.

The present invention relates to a male and female mating propagation method of tilapia, in specific to a mating propagation of tilapia which can adopt the pattern of male mating female; the method belongs to the technical field of aquatic product breeding. The method is characterized in that a net cage is equipped in an outdoor pond or indoor concrete pond; adult male tilapias are selected and people cut the upper jaw of the tilapias and put the tilapias into the net cage after being sterilized; and then adult female tilapias are selected to put in the net cage for mating culture and people can see the zygotes in the mouths of female tilapias after 7 to 10 days. With the present invention, a tilapia family line can be established for the family breeding of tilapias. In addition, people can clearly know the relation between the offspring and parent to control the inbreeding and avoid the germ plasm degeneration of offspring.

The invention relates to a method for using the frozen sperm of bass to induce the cultivation of sliding-tongue ovum thelykaryon, which comprises that: using the bass sperm frozen in the liquid nitrogen, to induce the thelykaryon cultivation of sliding-tongue ovum; defreezing the sperm in 37Deg.C water bath, taking 100muL sperm to dilute the MPRS for 10 times, to be arranged in the cultivate container at 6Deg. C; using 1000muj/cm2 ultraviolet to radiate for 0.2min, to inactivate the sperm; then using the inactive sperm to induce the fecundation with the ovum; in 5min of fecundation, using 2Deg. C water bath to shock the zygote, while the shock time is 20min; arranging the blast into the sea bath at 23Deg. C to be cultivated. The invention has low cost.

The invention discloses a method for fluorescent double-labeling microscopic observation of marine flounder oosperm microtubules and cell nucleuses. The method comprises the following steps of carrying out fixing of an oosperm with a membrane, stripping a blastoderm of the fixed oosperm from a whole embryo by a dissecting needle, punching the stripped blastoderm, reducing or eliminating the influence produced by uncombined aldehyde groups on an experimental result by sodium borohydride, carrying out sample sealing and fluorescent double-labeling dyeing, and carrying out sample observation by a fluorescence microscope or a laser scanning confocal microscope. The method has low sample requirements, has simple processes, is convenient for observation, produces a clear result, has a high three-dimensional feel, can be used for dynamic change research on flounder oosperm microtubules and cell nucleuses from blastoderm formation respectively to 2-cell, 4-cell and 8-cell development stages, is suitable for other larger telolecithal eggs of marine flounder oosperms, and has a wide application range.

The invention described herein provides high throughput methods and reagents for generating transgenic animals (e.g., mammals) through introducing materials into gametes or preimplantation stage (e.g., one-cell embryo or zygotes) via electroporation, leading to genetically inheritable modification to the genome of the animal.

The invention includes methods of enucleating avian eggs comprising visualizing internal structure of an avian egg utilizing TPLSM and ablating the nucleus of the egg by near-infrared light.

The invention discloses a kit and a method for constructing an apolipoprotein C2 (ApoC2) gene knockout hamster model, wherein the kit comprises sgRNA, which is obtained by PCR amplification of an artificial sequence as shown in SEQ ID NO: 2 and SEQ ID NO: 3, and in vitro transcription. The method comprises the following steps: designing the hamster ApoC2 gene specific targeting sequence, and preparing the cas9 mRNA and the sgRNA; Collection and culture of hamster fertilized eggs; Microinjection: co-injecting the sgRNA and cas9 mRNA into the cytoplasm of a hamster fertilized egg; Finally, the fertilized eggs were transplanted into the surrogate hamsters and the F0 generation hamsters were born. The ApoC2 gene knockout hamster model was successfully constructed by this method.

The invention relates to a cross method between large yellow Crocker and GuangDong large yellow Crocker. Wherein, their crossed generation has high grow speed and the application in long transmission; the invention selects GuangDong large yellow crocker as father, whose belly can be pressed to flow sperm; preparing dilute solution; compressing its belly to absorb sperm, whose activity is higher than 90% to be frozen and stored; mixing the dilute solution and anti-freeze agent, adding sperm, storing in frozen storage tube, sealing and putting in 4Deg. C; arranging it above the liquid nitrogen, reducing temperature and moving it into liquid nitrogen; selecteing the wide GuangDong large yellow crocker as mother; when its ovogen is mature, defreezing the sperm, and taking the freeze storage tube from liquid nitrogen to be put into room-temperature fresh water to be defrozen, activating it with seawater; artificial inseminating and washing zygote with seawater, removing sink zygote, and charging air and cultivating.

The invention provides a sturgeon zygote incubation device which comprises an incubation pool, a water supply system, a zygote placing screen and a gas supply system. The screen is placed in the cultivation pool. Sturgeon zygotes are placed into the screen. The screen is driven by water flow to rotate around a central station pipe of the cultivation pool. A gas pump power switch opens and closes a gas pump intermittently, and intermittent bubbles are formed on a gas stone. When the zygote placing screen in the cultivation pool rotates to the position where the gas stone is placed, the sturgeon zygotes in the screen are turned over under the effect of the bubbles, and the situation that due to long-time accumulation, hypoxia is caused, and the zygotes die can be prevented; meanwhile, as the gas pump power switch opens and closes the gas pump intermittently, the situation that due to continuous turning-over, the zygotes are injured can be avoided, and finally zygote incubation is finished. The sturgeon zygote incubation device has the advantages of being easy to manufacture and operate, high in practicality and the like.

The invention relates to an artificial breeding method of a new species of Murray cods. Conditions for culturing the new species in the source area Australia are simulated. The method includes the following steps that a temperature-controlled parent Murray cod breeding pond is built, a parent Murray cod nest is arranged, parent Murray cod breeding is carried out, parturition is hastened, artificial impregnation is carried out, zygotes are artificially hatched, and Murray cod fries are cultured; in this way, artificial breeding of the Murray cods is successful, and the artificial breeding method is an innovation in China.

The invention discloses an anhydrous transportation method for medaka zygotes, which relates to the technology of fish zygote transportation. In order to solve the technical problems, the invention provides the transportation method capable of keeping appropriate humidity and dissolved oxygen at the normal temperature so as not to affect the normal growth of the zygotes. The invention adopts the technical scheme that the method comprises the following steps of: selecting the zygotes of which fertilization time is 2 to 6 hours, removing whitish opaque dead eggs, flushing the zygotes by using Ringer's culture solution and then draining the water, filling the zygotes into a 2-5mL centrifuge tube provided with a plastic flat bottom and a cover and filled with solid oxygen beads to ensure thatthe total volume of the zygotes is about 4/5 of that of the centrifuge tube, covering the cover and then sealing the centrifuge tube by using a sealing tape, filling the sealed centrifuge tube into an A4-sized kraft envelope bag, and then putting the envelope bag into an EMS envelope to perform sealed transportation. The method is suitable for quickly transporting the medaka zygotes in an expressmode.

A method and vector system for delivering of a gene into a human stem cell for therapeutic uses is disclosed. The method includes linking a therapeutic gene to human sperm cells through a linker and fertilizing a human oocyte. The resulting zygote may then be cultured and established as human embryonic stem cells, which may later be differentiated into different types of cells for transplantation into the human body.

The invention relates to mutants of a fatty acid adenosine monophosphate (AMP) ligase and application of the mutants, and belongs to the field of bioengineering. The fatty acid AMP ligase comes from streptomyces roseosporus. The 296th glycine of the fatty acid AMP ligase is changed into alanine, and the 302th isoleucine of the fatty acid AMP ligase is changed into valine to respectively obtain the mutants pAL296 and pAL302. The mutants of the fatty acid AMP ligase can improve the binding level with exogenous precursor decanoic acid to enhance the yield of daptomycin. According to the invention, an encoding gene dptE of the fatty acid AMP ligase is subjected to site-directed mutagenesis in vitro by using computer simulation, overlap extension PCR and site-directed mutagenesis technologies to screen out the mutants pAL296 and pAL302 of which the affinities are obviously improved, recombinant shuttle plasmids pAK296 and pAK302 are constructed and sequentially led to the streptomyces roseosporus, and zygotes are screened to obtain streptomyces roseosporus DCE296 and DCE302 of the fatty acid AMP ligase with improved substrate affinities to lay a foundation for improving the production level of daptomycin.