16 results about "Sexual maturity" patented technology

The invention relates to the technical field of gene engineering, in particular to a transgenic zebrafish system pTo12-efla-tjp1a-P2A-mCherry based on a Tol2 transposition subsystem and a construction method of the transgenic zebrafish system pTo12-efla-tjp1a-P2A-mCherry. The preparation method comprises the following steps: S1, constructing a Tol2 transposon expression vector pTol2-ef1a-tjp1a-P2A-mCherry which contains an ef1a promoter, a tjp1a gene, a P2A peptide sequence and an mCherry reporter gene; s2, carrying out in-vitro transcription to prepare Tol2 transposase mRNA; s3, mixing the expression vector with Tol2 transposase mRNA, and microinjecting the mixture into the single-cell stage embryo of the zebra fish; s4, performing fluorescence screening on the F0 generation embryos surviving after injection to obtain the Founder fish with positive transgenosis; s5, after the F0-generation positive fish is bred to be sexually mature, an F1 generation is obtained through mating, transgenic positive individuals with stable inheritance are screened out after identification, and a transgenic zebrafish strain is established. According to the transgenic zebrafish system pTo12-efla-tjp1a-P2A-mCherry based on the Tol2 transposon system and the construction method of the transgenic zebrafish system pTo12-efla-tjp1a-P2A-mCherry based on the Tol2 transposon system, efficient integration of exogenous genes is achieved by utilizing the Tol2 transposon system, and the transgenic zebrafish line capable of being stably inherited to the F1 generation is successfully obtained.

This invention discloses a method for large-scale breeding of all-male salmon and trout, belonging to the field of fish sex regulation breeding technology. The method includes: collecting fertilized eggs from different families of the same species of salmon and trout, artificially incubating them to obtain fry, and inducing sex reversal in the fry by using 17β-estradiol when they begin feeding; subsequently, raising the fry to sexual maturity, selecting individuals with a female phenotype, and then using sdY as a sex identification gene, selecting pseudo-females with genotype XY by quantitative real-time PCR; pairing the sexually mature pseudo-females with genotype XY with normal males with genotype XY, and obtaining super-males with genotype YY through artificial insemination; pairing the sexually mature super-males with females with genotype XX, and producing all-male salmon and trout with genotype XY on a large scale through artificial insemination. This invention overcomes the problem that existing technologies cannot achieve large-scale breeding of all-male salmon and trout.

The invention discloses a method for promoting ovulation of sexual maturity zebrafish by using Gap27, and relates to the technical field of artificial breeding of fish, a gap connecting protein inhibitor Gap27 is applied to ovulation induction of zebrafish, an intercellular communication network in follicles is accurately intervened, the problem of asynchronous ovulation is solved, and in the final stage of maturation of oocytes, ovulation of sexual maturity zebrafish is promoted. Close substance and information exchange is carried out among follicular granule cells through a gap connecting channel formed by connecting protein Cx43, and Gap27 is used as a specific peptide fragment, can simulate an extracellular ring structure of Cx43, and competitively binds with Cx43 on an adjacent cell membrane, so that formation and functions of the gap connecting channel are physically blocked, and the follicular granule cells are separated from each other. When ovarian tissues of sexually mature female fish are exposed in a Gap27 solution, the transmission of inhibitory signals from granular cells to oocytes can be effectively interrupted, the rupture of nuclear membranes of the cells is promoted, and the final maturation process is completed.

The application belongs to the technical field of animal model construction, and relates to a construction method and application of a zebrafish aging model. The construction method comprises the following steps: 1, sgRNA, Cas9 protein and a homologous recombination knock-in plasmid containing fluorescent protein A and a knock-in gene are transferred into zebrafish embryos, development is cultured to sexual maturity, F0-I is obtained, and a knock-in strain I is obtained through screening; 2, sgRNA, Cas9 protein and a homologous recombination knock-in plasmid containing fluorescent protein B and a knock-in gene are transferred into zebrafish embryos, development is cultured to sexual maturity, F0-II is obtained, and a knock-in strain II is obtained through screening; 3, the knock-in strain I and the knock-in strain II are mated, zebrafish with fluorescent protein A and fluorescent protein B simultaneously expressed in the heart are screened from the offspring, and a wrn gene knockout homozygote, that is, a zebrafish aging model, is obtained. The construction method is more convenient to breed, significantly reduces breeding costs and the workload of screening and breeding, and has good application value.

The application discloses application of an IRS1 gene promoter region SNP site as a pig sexual maturity molecular genetic marker. The present application takes the IRS1 gene promoter region as a research object, and studies the correlation between the IRS1 gene promoter region and sow sexual maturity by using molecular and cell biology methods: it is found that g.128307611C>A and g.128307308C>T are significantly correlated with the age of sexual maturity of Duhei pig populations; then, the influence of different genotypes of the above sites on the activity of the IRS1 gene promoter is studied by transfecting different vectors. Further, the sow ovarian granulosa cells are taken as experimental materials, and experimental techniques such as transcriptome sequencing, total iron colorimetry and malondialdehyde colorimetry are used to study the influence of the IRS1 gene on the transcription level and ferroptosis of the sow ovarian granulosa cells, and it is found that the IRS1 gene influences the different genes of the cell transcriptome, and is mainly enriched in signal pathways such as iron ion homeostasis, and inhibits the ferroptosis of the cells.

The invention relates to the technical field of biological feed processing and application, and discloses an artificial feed for monochamus alternatus adult beetles as well as a preparation method and application of the artificial feed. The artificial feed provided by the invention can ensure that the adult monochamus alternatus hope has longer survival time and higher survival rate under the condition of not using a large number of fresh pine branches; meanwhile, by adding the female moth powder and the goose egg yolk powder, lecithin and other components can be introduced, the sexual maturity time of the adult monochamus alternatus can be remarkably shortened, the egg laying amount of the adult monochamus alternatus can be increased, and the breeding efficiency of the adult monochamus alternatus can be remarkably improved; in addition, the components in the artificial feed are reasonably matched, and the problem of feed hardening caused by addition of high wood chips is effectively avoided.

The invention provides a gene influencing sexual precocity of deep-county pigs and a screening method thereof, and belongs to the field of research on sexual maturity of pigs, the gene influencing sexual precocity of deep-county pigs comprises an SRD5A1 gene and / or a CYP11B2 gene, testicular tissues of deep-county pigs before and after sexual maturity are subjected to transcriptome sequencing, differential expression gene functions are analyzed, and the sexual precocity of deep-county pigs before and after sexual maturity is obtained. And screening out candidate genes related to the sexual maturity of the deep county pigs. And then performing transcriptome sequencing on testis tissues of the sexually mature deep county pig and the hybrid pig of the sexually mature deep county pig and the large white pig, analyzing functions of differential expression genes, and screening specific expression genes of the sexual maturity of the deep county pig. Integrating the results of the two times of transcriptome sequencing, analyzing the functions of the coincident genes, and screening candidate genes SRD5A1 and CYP11B2 which influence the sexual precocity of the pigs in the deep county according to the gene functions; and a foundation is laid for subsequent research on precocious puberty of pigs in deep county.

The invention discloses a method for creating fluorescent zebra fish based on an enhancer of a muscle specific gene. The invention provides a method for creating fluorescent zebrafish, which comprises the following steps: by taking an upstream promoter and an enhancer of a fmyhc2.2 gene as target sequences, amplifying the upstream promoter and the enhancer and seamlessly connecting the upstream promoter and the enhancer with a fluorescent protein carrier to form a recombinant plasmid, carrying out microinjection on a zebrafish embryo, selecting F0 generation individuals capable of driving fluorescent protein expression, and culturing until sexual maturity, thereby obtaining the fluorescent zebrafish. Selecting male and female individuals for selfing to obtain F1 generation, observing and selecting individuals with strong fluorescence from F1 generation embryos through a fluorescence microscope, and culturing the individuals with strong fluorescence to sexual maturity to obtain the transgenic zebrafish with remarkably discolored skin. The fluorescent zebra fish is created by utilizing the enhancer of the muscle specific gene, an innovative method is provided for creating a zebra fish ornamental strain, a support is provided for researching a regulation mechanism of zebra fish muscle related genes, and the fluorescent zebra fish has important scientific value and industrial application value.

The invention discloses a method for producing all-male largemouth micropterus salmoides based on soaking induction of sex reversal of largemouth micropterus salmoides, and particularly relates to the field of artificial propagation of all-male largemouth micropterus salmoides, the method comprises the following steps: S01, taking fairy shrimps as initial feed when largemouth micropterus salmoides fries obtained by incubation swim flatly; s02, transferring the largemouth micropterus salmoides fry into a water body in which a sexual reversal induction reagent is dissolved, keeping feeding three times every day during the period, and soaking and breeding the fry for 60 days; and S03, after soaking and culturing for 60 days, transferring the largemouth bass fry into a conventional culture water body for culturing, and after sexual maturity of the largemouth bass fry, obtaining a male population capable of generating seminal fluid, namely the all-male largemouth bass. According to the method, largemouth bass fries are soaked in a sex reversal induction reagent with a specific concentration, sex reversal fishes with a high survival rate and a low malformation rate can be directly obtained, and then large-scale cultivation of all-male largemouth bass is achieved.

This invention provides a method for rapid propagation of superior grass carp families, belonging to the field of aquaculture and fish genetic breeding technology. The method includes: screening for disease-resistant superior families from established grass carp families; isolating and enriching reproductive stem cells from these superior families; transplanting the grass carp family reproductive stem cells into the abdominal cavity or genital ridge region of recipient fish with short sexual maturity cycles; cultivating the recipient fish to sexual maturity, producing sperm, and then fertilizing the sperm with grass carp eggs to obtain F1 generation offspring; performing molecular identification on the offspring to confirm that their genetic material originates from the donor family. This invention can shorten the propagation cycle of superior grass carp families from 4-5 years to approximately 1.5 years, achieving precise and efficient transfer of superior traits, and providing an effective technical means to shorten the grass carp breeding cycle and cultivate superior grass carp varieties.

The application discloses a high-body cobia broodstock feed containing Chinese herbal medicines and a high-body cobia breeding method using the feed, and by matching different raw materials, the high-body cobia broodstock feed can better provide the high-body cobia broodstock with nutrient components and active substances. Before spawning, the high-body cobia broodstock feed can enhance the physical fitness and immunity of the broodstock, promote the development of the broodstock, accelerate sexual maturation, improve the spawning rate, the egg production, the fertilization rate and the hatching rate, and after spawning, the high-body cobia broodstock feed can promote the healing of the broodstock, restore the physical fitness, and guarantee the survival rate and quality of the broodstock after spawning.

The application relates to the field of aquatic organism measuring equipment. A device for rapidly identifying Euphausia superba and measuring biological characteristic parameters comprises a main box body, the top of the main box body is a measuring plate, at least three hollowed-out areas are arranged on the measuring plate in sequence, and a scale unit is arranged on the hollowed-out areas; the scale unit comprises a base plate, and a reference scale line is arranged on the upper side of the base plate; a slide rail with a left-right direction as a guide direction is arranged at the rear of the main box body; the device further comprises a sliding support which is slidably connected with the slide rail, and a camera and a tablet computer are arranged on the sliding support and connected with the tablet computer. The camera can take photos of the Euphausia superba, and then biological characteristics such as the body length, gender, sexual maturity, body color and feeding intensity of the Euphausia superba are recorded. The sliding support facilitates the sequential recording of the Euphausia superba on different scale units.

This invention discloses a method for constructing a zebrafish model of fatty liver based on glucokinase overexpression and its application. The method includes the following steps: using cDNA as a template, a zebrafish GCK gene fragment is obtained by PCR amplification; the vector fragment and the GCK gene fragment are homologously recombined using a homologous recombinase to obtain a recombinant plasmid; the recombinant plasmid is transformed into competent cells, and an endotoxin-free plasmid is extracted as a zebrafish GCK expression vector; a microinjection system is used to inject the plasmid into one-cell-stage fertilized eggs; after the fertilized eggs hatch into juveniles, fluorescent juveniles are selected for further culture until sexual maturity; sexually mature males and females are paired to spawn, and a stably heritable GCK-overexpressing zebrafish strain is selected from the offspring. This invention can obtain a zebrafish strain with stable high GCK expression, which can be used as a zebrafish model of fatty liver based on GCK overexpression.

The invention provides an RNA (Ribonucleic Acid) molecular marker for judging early gonad differentiation of young river snails in China and an identification method. Dmrt1 and Foxl2 genes are important genes related to gonad differentiation of river snails. The method comprises the following steps: designing and verifying primers SEQ ID NO: 1-2 according to conserved regions of Dmrt1 and Foxl2 genes; the method comprises the following steps: collecting gonad tissue samples in a young snail stage and a sexual maturity stage, wherein n is greater than or equal to 120, extracting total RNA (Ribonucleic Acid) of a field snail tissue, carrying out reverse transcription to obtain cDNA (Complementary Deoxyribonucleic Acid), detecting a double-gene expression quantity through qRT-PCR (Quantitative Reverse Transcription-Polymerase Chain Reaction), calculating a standardized expression value by taking beta-actin as an internal reference, and establishing a discrimination function D = log2 (Dmrt1 / Foxl2); 1, judging that the juvenile snails of the field snails start gonad differentiation; according to the invention, the problems of deficiency of specific molecular markers for sex differentiation of the young escargots in China and indefinite gonad differentiation time point are solved in a breakthrough manner, accurate locking of the gonad differentiation time of the young escargots is realized, the gonad differentiation time is 15-20 days earlier than morphological differentiation, and verification of more than one hundred of samples shows that the identification accuracy reaches 100%.

The invention belongs to the field of biological identification, and particularly relates to an identification method of a species telomere extension mechanism. Comprising the following steps: collecting a whole blood RNA sample of an adult individual after sexual maturity of a target species; detecting the expression condition of telomerase TERT in the whole blood RNA of the species, if the telomerase TERT is expressed, determining that the species are TERT positive telomere extension species; if the telomerase TERT is not expressed, continuing to identify; performing positive selection analysis on the adaptability of the alternative telomere extension ALT pathway related genes, and if the alternative telomere extension ALT pathway related genes are positive selection and are remarkably enriched in the pathway, determining that the alternative telomere extension ALT pathway related genes are ALT telomere extension related species; if the species are not positively selected and remarkably enriched, the species are TERT negative species. According to the invention, an identification method is provided for the telomere extension mechanism of species by detecting the expression condition of telomerase TERT in somatic cells of adult individuals of the species and performing positive selection analysis on alternative telomere extension ALT pathway related genes.

The invention provides an identification and analysis method for breeding biological characteristics of schizothorax in a plateau area. The invention discloses a plateau region schizothorax fish breeding biological characteristic identification and analysis method. The method comprises the following steps: S1, sample collection and basic data acquisition; s2, fish gonad development staging and reproduction cycle determination; s3, accurate age identification and growth relation establishment; s4, primary sexual maturity identification and analysis; s5, determining breeding seasons and periods; s6, determining spawning types; s7, high-precision fish fecundity determination and analysis; s8, carrying out correlation analysis on the reproductive capacity model and the environmental factors; and S9, evaluating a population breeding strategy and vulnerability. The plateau region schizothorax breeding biological characteristic identification and analysis method provided by the invention has the advantages that complete process operation from sample treatment to ecological evaluation can be realized, and the problems of scattered methods, low data precision and disjunction with environmental factors in the prior art can be solved.