193 results about "Zygote" patented technology

The invention provides an enhancer RNA (Ribonucleic Acid) molecule MZGAe1 and application thereof, and relates to the technical field of biology. The invention provides an enhancer RNA (Ribonucleic Acid) molecule MZGAe1. The nucleotide sequence of the enhancer RNA molecule MZGAe1 is as shown in SEQ ID NO: 1; meanwhile, the invention further provides a specific sgRNA sequence for activating the molecule in a targeted manner and a recombinant vector of the specific sgRNA sequence. Expression of endogenous MZGAe1 of cells is specifically activated by adopting a CRISPR activation technology, and conversion of mouse embryonic stem cells to bicellular-like cells can be efficiently promoted. By providing a brand-new endogenous targeted MZGAe1 accurate activation tool, the efficient transformation of wild mouse embryonic stem cells to bicellular cells can be realized only under the condition of endogenous activation of single enhancer RNA molecule MZGAe1, the proportion is at least increased by 5%, the expression of a totipotent marker gene ZSCAN4 is activated, and the expression of the totipotent marker gene ZSCAN4 is promoted. And an efficient and specific brand-new research tool and scheme are provided for researching zygote genome activation, embryonic development early events and cell reprogramming.

The invention discloses a construction method and application of an NMD escape model of an Alport syndrome mouse. A non-human animal model carrying Col4a5 gene c.4432delG frame shift mutation is prepared on the basis of a CRISPR / Cas9 gene editing technology. The method comprises the following steps: co-injecting gRNA of a 49 exon of a targeted Col4a5 gene, homologous recombinant donor oligonucleotide containing c.4432delG mutation and Cas9 nuclease into a mouse fertilized egg, and carrying out embryo transplantation to obtain an F0-generation mutant mouse; a mutation site is verified by combining PCR (Polymerase Chain Reaction) with sequencing, and a stably inherited mutation line is established through two generations of breeding. Through verification, the model accords with pathological characteristics of the Alport syndrome, can stably simulate typical clinical manifestation and pathological characteristics of the human X-linked Alport syndrome, and can be used as an important tool for research of the Alport syndrome.

The invention discloses an image detection and evaluation method for an unfrozen form of a fertilized egg embryo. The method comprises the following steps: acquiring a time sequence image sequence after the fertilized egg embryo is unfrozen; performing image standardization processing on the time sequence image sequence to generate a standardized embryo image set; extracting multi-dimensional embryo quality parameters from the standardized embryo image set; wherein the multi-dimensional embryo quality parameters comprise a morphological static parameter set, a dynamic change parameter set and a texture feature parameter set; generating a blastocyst formation rate based on the multi-dimensional embryo quality parameters; outputting a third-level embryo quality evaluation result according to a comparison relationship between the blastocyst formation rate and a preset threshold interval; wherein the three-stage embryo quality evaluation result comprises an excellent embryo grade, a qualified embryo grade and an abnormal embryo grade. The method has the following advantages and effects: full-automatic and multi-dimensional quantitative evaluation of the thawing process of the unfrozen embryos is realized, so that the scientificity of embryo screening and the success rate of clinical pregnancy are improved.

The invention discloses an artificial cultivation method of a hybrid of epinephelus trifoliatus and epinephelus lanceolatus, which comprises the following steps: taking healthy epinephelus trifoliatus as female parent fish, and carrying out cultivation strengthening and artificial ripening to obtain strengthened female parent fish; healthy epinephelus lanceolatus is used as male parent fish, and sexual mature male parent fish is obtained through natural sexual maturity; the ovulation time and the spermiation time are consistent; collecting egg liquid and seminal fluid, inseminating to obtain fertilized eggs, and performing embryo incubation and illumination regulation and control cultivation after opening to obtain hybrid spots; carrying out authenticity identification on the hybrid spots by utilizing a primer pair, and determining that the hybrid spots inherit genetic information of the female parent fish and the male parent fish; the primer pair is a sequence pair composed of SEQ ID NO. 1 and SEQ ID NO. 2, and a sequence pair composed of SEQ ID NO. 3 and SEQ ID NO. 4. The technical blank of crossbreeding by taking the epinephelus trifoliatus as the parent epinephelus is filled, and the authenticity of the hybridized spots is reliable.

The invention relates to the field of distant hybridization breeding, and discloses an artificial breeding method for hybrid species of humpback perches and epinephelus fuscoguttatus, which comprises the following steps: selecting healthy humpback perches as female parent fishes, and screening healthy epinephelus fuscoguttatus as male parent fishes; carrying out intensive cultivation on the parents by means of nutrient enrichment and temperature regulation and control, and promoting synchronous development and maturation of gonads of the parents; after the parents reach the breeding state, eggs of the female parent fishes and seminal fluid of the male parent fishes are collected respectively, fertilized eggs are obtained through artificial insemination, incubation management such as temperature control and continuous oxygenation is conducted on the fertilized eggs, and finally hybrid grouper fries are obtained. The crossbreeding application of the humpback perch as the female parent and the epinephelus fuscoguttatus is realized, and the problems that in the traditional hybridization process, parent gonad development is asynchronous, and filial generations cannot develop are effectively solved; the bred hybrid provides technical guarantee for standardization and precision promotion of grouper crossbreeding, and creates favorable conditions for breeding new hybrid lines with excellent characters of parents.

The invention relates to the technical field of fishery breeding, and particularly discloses a fry hatching device for schizothorax breeding, comprising a support frame, the top of the support frame is fixedly connected with a hatching box and a breeding box, the side surface of the hatching box is fixedly connected with the side surface of the breeding box, and two sides of the inner wall of the breeding box are fixedly connected with plastic plates; the fry hatching equipment comprises a hatching box and a breeding box, the side faces of the hatching box and the breeding box communicate with water adding pipes, the bottom of the side face of the hatching box penetrates through and is fixedly connected with a purification assembly, and an inlet is formed in the side, close to the hatching box, of the inner wall of the breeding box. And the aeration device is arranged, so that the content of dissolved oxygen in the incubator can be increased, sufficient oxygen is provided for fertilized eggs, and healthy growth of the fertilized eggs is promoted.

The invention provides a method for overspeed preparation of gene edited goldfish. The method comprises the following steps: S1, obtaining gobiocypris rarus without primordial germ cells as receptor fish; s2, designing a target spot for the goldfish amh gene, introducing a Cas9 / gRNA complex into a goldfish fertilized egg, performing sequencing verification to obtain a mutant individual, and feeding the mutant individual as a donor fish; s3, taking gonads of donor fish, cutting into pieces, performing enzymolysis digestion, filtering, and performing density gradient centrifugation to enrich donor germline stem cells; s4, transplanting the germline stem cells of the donor fish into the receptor fish body, and feeding to obtain the gene edited goldfish sperms and ova. S5, hybridizing and feeding goldfish amh mutant sperms and ova produced by gobiocypris rarus to obtain gene edited goldfish individuals. According to the invention, gobiocypris gobiocypris is used as a receptor, the cross-subfamily gene edited goldfish is used as a donor, and an efficient, stable and easy-to-popularize method for preparing the gene edited goldfish is provided by utilizing a belly borrowing reproduction technology.

The invention provides an artificial propagation method of Sinilabeo wuyi, and belongs to the technical field of aquatic animal breeding. The artificial propagation method of the Sinilabeo wuyi comprises the steps of parent fish classification, parent fish rearing, artificial spawning induction, natural insemination, artificial insemination, fertilized egg incubation, fry rearing, juvenile fish rearing and the like. The oxytocic is reasonable in compatibility, the injection time of the oxytocic is accurate, the oxytocic rate, the fertilization rate and the hatching rate are increased, and damage to parent fishes is small. According to the method, feeding methods in different stages of fry breeding are also defined, the survival rate of the sinilabeo wuyi fry breeding stage is increased, and a technical support is laid for artificial breeding of the endangered species sinilabeo wuyi.

The present invention addresses the problem of specifically clarifying the effects on a fertilized egg when 5-ALA is administered to a human female suffering from infertility, and constructing a more effective means for treating infertility through the suppression of chromosomal abnormalities. It has been confirmed that, when 5-ALA is ingested in combination with a drug therapy that is generally performed in the treatment of female infertility, the rate of chromosome aneuploidy in a fertilized egg or the like is significantly reduced, and the qualitative improvement of an embryo through improvement of the stability of the chromosomes is expected.

The application discloses a submerged Cinnamomum micranthum in-vitro rapid propagation and brown-preventing seedling raising method based on zygote embryos, directly induces adventitious buds of the submerged Cinnamomum micranthum for the first time, significantly improves a propagation coefficient, and overcomes problems such as vitrification and browning of the adventitious buds of the submerged Cinnamomum micranthum. The application establishes an efficient and stable in-vitro regeneration system of the submerged Cinnamomum micranthum, and provides a basis for large-scale seedling raising of the submerged Cinnamomum micranthum.

The invention provides a culter turunx2b gene knockout method and application of the culter turunx2b gene knockout method in creation of intermuscular thorn-free mutants, and belongs to the technical field of gene editing. The invention provides an efficient gene knockout method which comprises the following steps: selecting two targets on a second exon of a culter turunx2b gene as target sites, designing and synthesizing two gRNAs, mixing the gRNAs with Cas9mRNA, introducing the mixture into a fertilized egg of culter turunx2b in an I cell stage in a microinjection manner, hatching, and screening mutated F0-generation culter turunx2b individuals. By utilizing the method disclosed by the invention, the F0-generation knockout efficiency is up to 87.5% under the condition of ensuring that the fertilized eggs are hatched to survive. The method disclosed by the invention is simple and easy to implement and simple to operate, the culter alburnus gene mutation strain can be rapidly obtained by utilizing the gene mutation method, and the method has important scientific significance for developing economic fish gene function research, revealing genetic development law and creating new germplasm.

The invention discloses a bactrocera dorsalis point mutation gene editing method based on CRISPR-Cas9 and homologous directional repair, and relates to the technical field of gene editing. The method comprises the following steps: designing high-specificity sgRNA for a PAM sequence adjacent region of a target gene; extending the length of the homologous arm to 100bp, and designing a single-stranded DNA (deoxyribonucleic acid) containing a mutation site as a homologous repair template; after a mixture containing the high-specificity sgRNA, the homologous repair template and Cas9 protein is injected into a bactrocera dorsalis fertilized egg in a microinjection mode, incubation and incubation are carried out, and G0-generation imagoes are obtained through culture; hybridizing the G0-generation imago with wild bactrocera dorsalis, and screening to obtain a G1-generation hybrid mutant; and carrying out selfing by using the G1-generation heterozygous mutant, and screening to obtain a G2-generation homozygous mutant. By adopting the method, the mutation efficiency and embryo survival rate of bactrocera dorsalis can be effectively improved.

This invention discloses a gRNA combination and its application in the preparation of drugs for the prevention of metabolic-associated fatty liver disease (MASLD). The gRNA combination, when mixed with Cas9 protein, yields an RNP complex, which, when microinjected into mouse zygotes, can breed a stable and heritable mouse strain with TMEM68 gene knockout. Combining lipidomics, transcriptomics, and primary hepatocyte functional verification, the core regulatory role of TMEM68 in hepatic lipid metabolism is revealed for the first time systematically. Experiments demonstrate that TMEM68 deficiency significantly reduces the storage of triglycerides (TAG) in the liver and hepatocytes, decreases lipid droplet formation, and reshapes the metabolic homeostasis of various lipids such as glycerophospholipids, cholesterol esters, and bile acids. Therefore, the gRNA combination of this invention, or the RNP complex obtained by mixing the gRNA combination with Cas9 protein, can be used to prepare drugs for the prevention of MASLD, providing a novel intervention strategy for MASLD prevention with broad application prospects.

The invention belongs to the technical field of GPCR drug research and development, and discloses a ratio type cAMP fluorescent probe RaCamp and a construction method of a transgenic mouse, the probe RaCamp is formed by series fusion of G-Flamp2 fluorescent protein and mCherry fluorescent protein through a flexible linker, and the preparation method of the probe RaCamp comprises the following steps: S1, obtaining an mCherry fragment; s2, a G-Flamp2 linear carrier is obtained; s3, homologous recombination; s4, converting and screening; the construction method of the transgenic mouse comprises the following steps: T1, designing a carrier; t2, microinjection of fertilized eggs; t3, embryo transplantation and reproduction; the invention has the beneficial effects that mCherry is added as an internal reference protein on the premise of keeping the sensitivity of the original G-Flamp2 probe, and the selected connecting peptide (GGGGS) 2 enables the expression quantity of the two proteins in cells to be accurately controlled to be 1: 1, so that the problem of background interference in cell observation of in-vivo imaging is well solved. And the sequence is inserted into a Rosa26-LSL box, so that the construction of a Rosa26-LSL-RaCamp mouse is realized.

The application relates to the field of aquaculture, and particularly discloses a method for cultivating oyster larvae, which comprises the following steps: S1, fertilized eggs are developed into D-shaped larvae in seawater at 22 DEG C for 20-24 hours; S2, the D-shaped larvae are cultivated under the condition that the temperature is 22-24 DEG C and the salinity is 29-31 ‰, feed and growth nutrient solution are fed daily, water is changed daily, after the D-shaped larvae appear eye spots, a larval collector is put in, and algal feed and growth promoting solution are fed daily, the larvae are cultivated into 0.8-1 mm juvenile shellfish, and are transferred to sea cultivation; the growth nutrient solution comprises the following components in weight parts: 1-2 parts of a small molecule peptide solution, 0.01-0.03 parts of neomycin sulfate powder, 0.5-1 part of corn starch, 2-4 parts of modified montmorillonite, and 10-15 parts of water. The method for cultivating oyster larvae has the advantages that the survival rate of the larvae is improved, the growth speed is accelerated, and the metamorphosis and attachment rate is improved.

The application belongs to the technical field of fish genetic breeding, and specifically discloses an artificial induction method of Megalobranchius ambius and Culter alburnus hybrid triploid and application. The artificial induction method comprises the following steps: S1. taking Megalobranchius ambius as a female parent and Culter alburnus as a male parent to perform artificial dry insemination; S2. performing hydrostatic pressure treatment on the fertilized eggs, and the hydrostatic pressure treatment conditions are as follows: the starting time of treatment after fertilization is 2-3 min, the pressure is 35-50 MPa, and the duration is 3 min; S3. hatching the fertilized eggs and performing transfer culture, and the water temperature in the whole process from fertilization, hydrostatic pressure treatment, hatching to fry cultivation is controlled to be 22-24 DEG C; and Megalobranchius ambius and Culter alburnus triploid is screened through ploidy analysis. The method can successfully induce Megalobranchius ambius and Culter alburnus hybrid sterile triploid with superior growth performance, and based on comprehensive index evaluation, the best hydrostatic pressure treatment condition is obtained, high triploid rate (>80%) and high survival rate (>98%) are simultaneously achieved, and commercialized breeding production can be realized.

The application provides a method for on-site rapid artificial insemination of wild Pseudosciaena crocea of Lutjanidae, and belongs to the technical field of aquatic animal breeding. Wild parent fish of Pseudosciaena crocea of Lutjanidae are captured in the breeding season; sperm is collected from the selected sexually mature male parent fish on the capture site; after artificial sperm collection, sexually mature female parent fish are selected, and fish eggs are collected; high-quality fertilized eggs are obtained through artificial insemination; the artificial insemination comprises the following steps: adding sperm into fish eggs, adding natural seawater for stirring, and finally washing with natural seawater. Through comprehensive regulation of key links in the whole process of artificial insemination, the purpose of obtaining high-quality fertilized eggs by using wild parent fish of Pseudosciaena crocea of Lutjanidae and carrying out artificial fry breeding is achieved.

The invention discloses an artificial breeding method of lota lota, which comprises the following steps: S1, selecting robust lota lota as parent fish, placing the parent fish in an indoor culture tank for temporary culture, and inducing gonad to develop mature by regulating and controlling water temperature; s2, performing artificial spawning induction on the parent fish with mature gonad to obtain ova and semen, and performing in-vitro fertilization on the ova and the semen to obtain fertilized ova; s3, performing two-step soaking treatment on the fertilized eggs by using a composite biological functional liquid containing a liquid A and a liquid B, and then transferring the fertilized eggs into a hatching facility for hatching to obtain fish fries; and S4, breeding the hatched fish fries. According to the artificial breeding method of the lota lota, the survival rate of fertilized eggs and the survival rate in the fry breeding stage are increased to a high level, large-scale and controllable artificial breeding of the lota lota becomes possible, and the overall technical scheme has high stability and industrial application value.

Methods for improving parthenogenesis efficiency by DWT1 and BABY BOOM transcription factors in plants are provided. A rice embryo trigger transcription factor BABY BOOM1 can initiate embryogenesis when expressed in the unfertilized egg cell through a process called parthenogenesis (Khanday et al., 2019. Nature 565: 91-95). The parthenogenesis efficiency by BABY BBOM1 itself is 10-29%. This invention describes methods of high frequency of parthenogenesis by simultaneous expression of BABY BOOM and DWT1 transcription factors. When BABY BOOM1 and DWT1 are expressed together through egg cell-specific promoters, parthenogenesis efficiencies of up to 90% are achieved. These high parthenogenesis efficiencies are a prerequisite for field applications of synthetic apomixis in crop plants.

The invention discloses a construction method of a COL4A5-K229X point mutation X-linked Alport syndrome mouse model. The construction method comprises the following steps: aiming at c.685Agt of a No.12 exon of a mouse COL4A5 gene; carrying out T point mutation, and designing and preparing Cas9 mRNA, gRNA and a donor vector; the components are mixed and then microinjected into fertilized eggs of a C57BL / 6J mouse to obtain an F0-generation mouse; identifying the genotype through PCR (Polymerase Chain Reaction) amplification and Sanger sequencing, and screening positive mice; mating the positive F0-generation mice with the wild-type mice, and breeding F1-generation and subsequent generations; the phenotype of the model is further verified through qPCR, biochemical analysis, light microscopic examination, transmission electron microscope and immunofluorescence. The model constructed by the invention shows hematuria, proteinuria, azemia, podocyte loss and irregular thickening and layering of glomerular basement membrane, is consistent with phenotypes of human XLAS patients, and provides an animal model tool for analyzing pathogenesis and developing treatment strategies.

The invention discloses a construction method of a Klf6 gene knockout mouse embryo model, which comprises the following steps: S1, preparing a gene editing solution containing Cas9 enzyme and sgRNA for targeting a third exon region and a fourth exon region of a mouse Klf6 gene by utilizing a CRISPR / Cas9 gene editing technology; s2, taking out the fertilized eggs of the mouse, transfecting the fertilized eggs of the mouse with a gene editing solution, and then performing in-vitro culture on the fertilized eggs; s3, carrying out embryo transplantation on the two-cell stage embryo obtained in the step S2 into a pregnant female mouse, and then carrying out conventional feeding; and S4, 7-8 days after embryo transplantation, taking out the uterus and stripping the embryo to obtain the Klf6 gene knockout mouse embryo model. According to the method, gene knockout is carried out by using CRISPR / Cas9 in the fertilized egg period in combination with an electrotransfection method, and compared with an existing method, the construction process time can be greatly shortened, the working efficiency is obviously improved, and the success rate is high.

The invention belongs to the technical field of transgenic and biomedical research tools, and particularly relates to a method for constructing a transgenic fish system based on a Tol2 transposon system, which comprises the following steps: constructing a myo6b-lhfpl5b-DsRed plasmid by using myo6b as a promoter, lhfpl5b as a target gene and DsRed as a red fluorescent protein tag through a microinjection method, and constructing a Tol2 transposon system-based transgenic fish system. Co-injecting the plasmid containing the Tol2 transposase mRNA and the Tol2 transposase mRNA into a fertilized egg of an AB zebra fish strain, and screening generation by generation to obtain a transgenic fish strain with stable inheritance MET channel gene lhfpl5b marked hair cells. The spatio-temporal dynamic visualization of the living whole fish to the lhfpl5b is realized, the deafness-causing molecular pathway is clarified, and an in-vivo platform is provided for drug screening and intervention strategies.

The invention discloses a method for efficiently producing triploid oysters, and belongs to the technical field of aquaculture breeding. The method comprises the following steps: S1, obtaining sperms and ova of diploid oysters, and carrying out sperm-ova mixed fertilization; s2, performing induction treatment on the fertilized eggs to inhibit release of a second polar body; s3, collecting fertilized eggs subjected to induction treatment, cleaning, and hatching to a larva stage; s4, 24-72 hours after fertilization, screening based on larva individual size differences, and collecting a target larva population; and S5, culturing the larvae screened in the step S4. According to the method provided by the invention, even if the initial induction rate is only about 70%, the larva population with the triploid rate higher than 90% can be stably obtained finally, the hatching rate and the survival rate are high, the method is easy and convenient to operate, and the requirement for induction conditions is low; the method is suitable for triploid preparation of precious oyster varieties in which a large number of fertilized eggs are difficult to obtain under strong induction conditions or oyster strains with important economic characters.

The application belongs to the technical field of biology and particularly relates to a functional recombinant Cas9 protein targeted to an ovocyte of Procambarus clarkii and application thereof. The protein is NLs-VgSP-Cas9-NLs. The application directly delivers the recombinant Cas9 protein, thereby reducing the risk of integration of exogenous genes. By specifically targeting the ovocyte, off-target effects on somatic cells can be avoided, the efficiency of editing of germ cells is improved, and early development or genetic manipulation can be conveniently studied. The ovocyte of Procambarus clarkii is large, the recombinant Cas9 protein is efficiently delivered by receptor-mediated endocytosis on the surface of the ovocyte, and the problem that microinjection of zygotes cannot be implemented is solved.

The invention belongs to the technical field of biology, and particularly relates to a construction method and application of a gobiocypris rarus gnaq knockout strain, a CRISPR / Cas9 gene editing technology is adopted, and double-target sgRNA (target 1 induces-8bp deletion and target 2 mediates + 32bp insertion) is designed for gobiocypris rarus gnaq genes. The sgRNA sequence is optimized through ZiFiT target spot design software, and the target specificity is ensured through NCBI BLAST comparison. The sgRNA and Cas9 mRNA which are transcribed in vitro are jointly microinjected into a gobiocypris rarus fertilized egg, and an F0 generation chimera is obtained. After the F1 generation heterozygote selfing, the homozygous mutants gnaq-8 and gnaq + 32 which conform to the Mendel genetic law are successfully screened in the F2 generation. According to the invention, gnaq knockout in gobiocypris rarus is realized for the first time, the blank of gobiocypris rarus G protein signal channel research is filled, and an important model is provided for subsequent fish related gene function research (such as antiviral immunity and tumor mechanism).

This invention discloses an indoor breeding device for largemouth bass and its application. The device includes a parent selection unit and a spawning unit. The parent selection unit includes a parent rearing pond and a parent collection net cage, with the parent collection net cage located in the parent rearing pond near the inlet. The spawning unit includes a spawning pond and fish nests within the pond, wherein the fish nests are separated to form relatively independent breeding spaces within the pond, and a water flow system is installed above the fish nests. The advantages are: (1) This invention can effectively improve the indoor breeding efficiency of largemouth bass, from difficulty in spawning to a breeding rate of over 40%; (2) The application method utilizes the largemouth bass's head-up phenomenon to observe the gonadal development of the parent fish and uses this to screen for parent fish with mature gonads, greatly improving efficiency and without harming the fish fry; (3) When the parent fish spawn, the device adaptively adjusts the water flow in the pond according to the first collection of fertilized eggs to ensure the breeding rate of the parent fish.

The invention discloses a construction method of a glomerular IgA deposition mouse model, and relates to the technical field of animal model development and glomerular IgA dyeing. Comprising the following steps: obtaining Cas9 mRNA (messenger ribonucleic acid), guideRNA (ribonucleic acid) and donor DNA (deoxyribonucleic acid) in an in-vitro transcription manner; the method comprises the following steps: carrying out microinjection on Cas9 mRNA (messenger ribonucleic acid), gRNA (guide ribonucleic acid) and donor DNA (deoxyribonucleic acid) into fertilized eggs of a C57BL / 6J mouse; carrying out genotype identification on the F0 generation of birth mice through PCR (Polymerase Chain Reaction) amplification and sequencing; mating the F0-generation positive mouse with a wild type C57BL / 6J mouse, and breeding to obtain an F1-generation mouse; and spontaneous glomerular IgA deposition appears in 24 weeks after the F1 generation heterozygote mouse is born. According to the method, accurate mutation is introduced by utilizing a homologous recombination repair mechanism to construct a glomerular IgA deposition model, the efficiency is high, and the off-target risk is low.

The utility model discloses a device suitable for hatching fertilized eggs of hatched fish, which comprises a water tank (1) and at least one hatching barrel (2), a water inlet header pipe (3) and a water return header pipe (4) are connected between the water tank (1) and the hatching barrel (2), and a water pump (5) is arranged on the water inlet header pipe (3); each hatching barrel (2) is respectively connected with the water inlet main pipe (3) and the water return main pipe (4) through a water inlet branch pipe (9) and a water return branch pipe (10) of the hatching barrel (2); a main body of the hatching barrel (2) is cylindrical, the bottom of the hatching barrel (2) is spherical, the central angle of the spherical bottom is 120-160 degrees, the height-diameter ratio of the barrel is 4-6, and a water outlet of a water inlet branch pipe (9) is right opposite to the center of the spherical bottom and is 2-3 cm away from the bottom. The device is suitable for hatching roes of various hatched fishes, is also suitable for hatching of partial half-hatched and non-hatched demersal roes, and is especially suitable for breeding work in small and medium-sized fishing grounds.

The invention provides a method for constructing an organ regeneration genetic compensation effect, and relates to the technical field of biology. Comprising the following steps: S1, designing sgRNA; s2, carrying out in-vitro transcription and purification to obtain a high-purity sgRNA solution; s3, preparing fertilized eggs, preparing sgRNA mixed injection, and performing embryo culture after micro-injection operation to obtain F0-generation animal larvae; s4, performing gene knockout efficiency detection on the F0-generation animal larvae, screening out the F0-generation animal larvae with high knockout efficiency, and breeding and mating the F0-generation animal larvae to obtain F1-generation animals; and S5, verifying and screening the F1-generation animals to obtain homozygous knockout individuals, carrying out amputation treatment on the homozygous knockout individuals, and verifying the genetic compensation effect of organ regeneration. The YAP knockout animal is constructed through a gene editing technology, the effect and mechanism of the YAP knockout animal in limb regeneration are researched, whether YAP deletion affects the regeneration capacity through a genetic compensation effect or not can be revealed, and then a theoretical basis and an experimental basis are provided for future regenerative medicine application.

The invention provides an artificial propagation method of schizothorax wangchizoides, and belongs to the technical field of fish propagation. The artificial propagation method of the schizothorax wandersoni comprises the following steps that (1) mature parent fish is selected and injected with an oxytocic mixture, and the oxytocic mixture comprises DOM, HCG and LRH-A3; (2) collecting ova and sperms after the parent fish is oestrus, and carrying out artificial insemination; and (3) transferring the fertilized eggs into an incubator for incubation. According to the method, 268345 artificial fertilized eggs are obtained, the fertilization rate reaches 95.6%, 251707 pond fries are obtained after artificial incubation, the hatching rate is 93.8%, and the fertilization rate and the hatching rate of the schizothorax wandersoni are remarkably increased.