35 results about "Capacitation" patented technology

The invention discloses a flow cytometry-based sperm acrosome reaction detection reagent which comprises acrosome reaction induction fluid and staining fluid, as well as sperm capacitation fluid, the pH value of the sperm capacitation fluid is 7.0-7.8, the sperm capacitation fluid contains sodium chloride, potassium dihydrogen phosphate, glucose, potassium chloride, magnesium sulfate heptahydrate, calcium chloride dehydrate, N-2-piperazine-N-2-ethanesulfonic acid disodium salt, sodium bicarbonate, sodium acetone, bovine serum albumin, and sodium lactate solution, the sperm capacitation fluid replaces the current sperm culture fluid, so that the capacitation effect is good and the induction is facilitated; the sperm acrosome reaction can be detected by using flow cytometry, and the flow cytometry-based sperm acrosome reaction detection reagent is accurate in the sperm acrosome reaction detection result, stable and reliable, good in repeatability, and clinically easily popularized.

The invention relates to a horizontal-shaft tidal turbine with linkage spiral vanes. The horizontal-shaft tidal turbine comprises a spindle, a case, a thrust bearing, a coupling, a generator, a rack, a base and a water guide cone, wherein the lower side of the case is respectively supported by the rack and the base; the horizontal-shaft tidal turbine is characterized by further comprising linkage spiral vanes in two-dimensional linkage spiral line shape, a cone-like-shaped hub immersing into tide, and a sealing cover; the generator is connected with the spindle and the cone-like-shaped hub by the coupling, the sealing cover is arranged at the junction of the spindle and the cone-like-shaped hub, the water guide cone is arranged at the tail part of the case at the rear side of the cone-like-shaped hub; and the linkage spiral vanes rotate at uniform speed at the periphery of the cone-like-shaped hub by forming gradient stretching along the axial direction of the cone-like-shaped hub to form a spatial warping shape to be uniformly distributed via the thrust bearing. The horizontal-shaft tidal turbine can greatly reduce hydraulic friction collision and reduce loss at the entrance head, thus improving the capacitation effect of the water turbine and well solving the difficulty of allowing ocean fish schools to smoothly pass.

The invention provides a method for detecting neuraminidase influencing sperm functions. The method is a novel laboratory detecting method for assessing quality and functions of a sperm and can provide detection and clinical consultations for male infertility patients without clear clinical reasons for infertility. The method is characterized in that expressions of neuraminidase in the sperm are visually quantized through single sperm fluorescence strength analysis or are visually observed by means of fluorescence imaging under a microscope, and if the neuraminidase is highly expressed in the sperm, capacitation of the sperm is achieved through external capacitation liquid, and neuraminidase activity is detected in a fluorescence method.

This disclosure provides a method for determining male fertility status. The method comprises determining GM1 localization patterns following induced sperm capacitation, identifying the percentage of various patterns, particularly the ratio of [(AA+APM) / total number of GM1 localization patterns] and determining if the percentage of certain GM1 localization patterns in response to induced capacitation is altered. Based on the change in the percentage of localization patterns of certain patterns in response to induced capacitation, alone or in combination with other sperm attributes, male fertility status can be identified.

The diagnosis of male infertility is based predominantly on the results of standard semen analysis for concentration, total motility, progressive motility, volume, pH, viscosity and / or morphology. When sperm enter the female reproductive tract, they must undergo a series of physiological changes, known as capacitation, in order to fertilize an egg. This process involves plasma membrane changes that occur in response to stimuli within the female tract. These changes include removal of sterols and redistribution of the ganglioside GM1. Semen analysis identifies only half the cases of male infertility due to standard semen analysis providing little information on sperm functional competence. Previous data demonstrated that localization of the ganglioside, GM1, identifies sub-populations of sperm capable of undergoing the functional maturation process known as capacitation and tracks strongly with fertility.

This invention relates to a method of producing an auto induced biological barrier against spermatozoids and / or other microorganisms including viruses and / or pre cancers or non invasive cancers involving the production of an autoimmune reaction in a host in need of such auto-inducement. The invention is characterized in that it comprises the steps of producing heat shock proteins inside vaginal and cervical mucosal cells and releasing said heat shock proteins into the vaginal cavity thus inducing an autoimmune response through a trigger and / or stimulation of both innate and adaptive systems and, as a consequence, the production of: a contraceptive effect through the elimination of the migration or free movement of individual or populations of spermatozoids and / or an inhibition of the maturation and / or capacitation process of the same and / or a neutralization or destruction of the same; a prophylactic effect through an elimination of the migration or free movement of individual or populations of other microorganisms including viruses and / or a neutralization or destruction of the same; a prophylactic effect against pre cancers; a therapeutic effect against non invasive cancers. The invention also relates to a device for using said method comprising a hollow body (1) provided with a heat source on the inside as well as with a means (13, 27) of removal of said hollow body from the vaginal or rectal cavity and a means (6, 8, 10) capable of exchanging heat originating from the hollow body.

The invention relates to a horizontal-shaft tidal turbine with logarithmic spiral vanes. The horizontal-shaft tidal turbine comprises a spindle, a case, a thrust bearing, a coupling, a generator, a rack, a base and a water guide cone, wherein the lower side of the case is respectively supported by the rack and the base; the horizontal-shaft tidal turbine is characterized by further comprising logarithmic spiral vanes in two-dimensional logarithmic spiral line shape, a cone-like-shaped hub immersing into tide, and a sealing cover; the generator is connected with the spindle and the cone-like-shaped hub by the coupling, the sealing cover is arranged at the junction of the spindle and the cone-like-shaped hub, the water guide cone is arranged at the tail part of the case at the rear side of the cone-like-shaped hub; and the logarithmic spiral vanes rotate at uniform speed at the periphery of the cone-like-shaped hub by forming gradient stretching along the axial direction of the cone-like-shaped hub to form a spatial warping shape to be uniformly distributed via the thrust bearing. The horizontal-shaft tidal turbine can greatly reduce hydraulic friction collision and reduce loss at the entrance head, thus improving the capacitation effect of the water turbine and well solving the difficulty of allowing ocean fish schools to smoothly pass.

The invention discloses a mouse cryopreserved sperm resuscitation liquid compound and a preparation method and application thereof. The compound comprises a basis solution, methyl B cyclodextrin MBCDand polyvinyl alcohol PVA. The preparation method comprises the steps that firstly, methyl B cyclodextrin MBCD and polyvinyl alcohol PVA are dissolved in the basic solution, after sufficient dissolution, the mixture is filtered through a bacterium filter device, and mouse cryopreserved sperm resuscitation liquid is obtained. According to the compound, methyl B cyclodextrin MBCD and polyvinyl alcohol PVA are added to the sperm resuscitation liquid, the sperm capacitation process is promoted, the capacitation of sperms is improved, the number of the sperms with fertilization capacity is increased, the fertilization probability of the sperms and eggs is increased, and the fertilization rate is increased. The compound is beneficial to successful preservation of mouse sperm resources, and reestablishment of mouse strain resources is increased and accelerated.

The invention discloses a spermin-vitro capacitation solution, a kit applied to in-vitro fertilization and an in-vitro fertilization method of mammals. The spermin-vitro capacitation solution comprises 110-140mmol of NaCl, 2.40-2.95mmol of KCl, 22.5-27.5mmol of NaHCO3, 4.8-6.0mmol of glucose, 0.22-0.30mmol of sodium pyruvate, 0.22-0.30mmol of taurine, 0.04-0.06mmol of epinephrine, 14-16mg / ml of BSA, 0.4-0.6mmol of MgCl2, 1.6-2.0mmol of CaCl2 and 8-12mmol of a DL-sodium lactate solution, wherein the pH value of the spermin-vitro capacitation solution is 7.35+ / -0.10. The spermin-vitro capacitation solution is capable of obviously improving the in-vitro fertilization efficiency of mongolian gerbil; after the method is used, the in-vitro fertilization efficiency of mongolian gerbil is greatlyimproved; two-cell development rate reaches 65% or above; the maximum two-cell development rate can reach up to 85%; the in-vitro fertilization rate obtained by the method is equivalent to that of a microinjection method and is even higher than that of the microinjection method; the method is convenient to use and has a good application prospect.

Belonging to the technical field of livestock breeding, the invention discloses a bovine sperm capacitation solution and an in vitro sperm capacitation method. The bovine sperm capacitation solution mainly consists of the following components: 6.7500mg / mL NaCl, 0.3000mg / mL KCl, 2.5000mg / mL NaHCO3, 0.0620mg / mL NaH2PO4.2H2O, 0.2960mg / mL CaCl2.2H2O, 0.1000mg / mL MgCl2.6H2O, 0.1120mg / mL sodium pyruvate, 0.0186mg / mL EDTA-Na, 0.0100mg / mL phenol red, 6.0000mg / mL BSA, 1.0000mg / mL glucose, 0.4800mg / mL caffeine, 0.0500mg / mL heparin, and 0.025-0.05mg / mL Nisin. The capacitation solution provided by the invention can effectively ensure the sperm motility, sperm plasma membrane integrity rate, acrosomal integrity rate and mitochondrial activity, and maintains the in vitro sperm capacitation effect.

The invention discloses simple and high-efficient method and device for separating sperms. The device consists of an Eppendorff centrifugal tube, a separating tube and a rubber ring. The method comprises the steps of: reasonably controlling the level height of culture solution; and culturing in a 5 percent CO2 culture container at 37 DEG C for 15minutes for separating the sperms. The concentration of the sperms can be adjusted by adopting a low-speed centrifugal method; about 95 percent of obtained sperms have athletic ability; and the collected solution generally has no blood cells. The invention is suitable for an in vitro capacitation experiment.

The invention provides a method for improving the sperm motility and increasing an in-vitro-fertilization rate of frozen bovine semen by the aid of genistein. The method is particularly characterized in that the genistein containing 1%o of DMSO (dimethyl sulfoxide) is added into SP-TALP (serum phosphate-total alkaline phosphatase) washing liquid for washing the frozen bovine semen in earlier-stage processing for the frozen bovine semen, or mixed liquid of PHE (phenylalanine) capacitation liquid and IVF-TALP (in vitro fertilization-total alkaline phosphatase) fertilization liquid containing bovine semen in a capacitation processing stage of second-stage pre-fertilization semen or prepared fertilization drops containing oocytes in a third-stage in-vitro-fertilization procedure; the added amounts of the genistein in two earlier stages of a fertilization procedure of each bottle of frozen bovine semen are 10 micromoles per liter, and the final concentration of the genistein in mixed liquid of the bovine semen and the fertilization drops containing the oocytes is 10 micromoles per liter when the genistein is added in a third stage of the fertilization procedure of each bottle of frozen bovine semen. The method has the advantages that the genistein is respectively used in the three in-vitro-fertilization stages, so that the sperm motility is greatly improved, and the sperm fertilization rate is greatly increased.

The invention relates to a method for in vitro capacitation of rabbit sperms and a nutrient solution formula. In the invention, 10 mum mol / L of EGCG is added in a BO capacitation liquid for sperm in vitro capacitation treatment, and 10 mum mol / L of EGCG is added in the BO capacitation liquid, the formula of the a BO capacitation liquid comprises the following components by weight: 6.5453mg / mL of NaCl, 0.2997mg / mL of KCL, 0.1145mg / mL of NaH2PO4.H2O, 0.1057mg / mL of MgC12.6H2O, 0.330mg / mL of CaCl2.2H2O, 2.518mg / mL of glucose, 4% of 0.2% phenol red, 100IU / mL of penicillin, 100IU / mL of streptomycin, 3.1080mg / mL of NaHCO3, 0.1380mg / mL of sodium pyruvate, 20mum g / mL of heparin and 20mg / mL of BSA. The method for in vitro capacitation of rabbit sperms overcomes the defect of mass oxyradical accumulation generated by sperm metabolism due to large sperm density during a current sperm in vitro capacitation process. The method uses adult New Zealand white male rabbit, different concentration EGCG (0mumM, 10mumM, 15mumM, 20mumM and 25mumM) can be added in the BO capacitation liquid, influence of the BO capacitation liquid to the rabbit sperm capacitation effect is researched so that optimum EGCG addition concentration is obtained, and thereby a rabbit in vitro capacitation culture system with high efficiency is established.

The invention discloses a low-dosage high / low density gradient centrifugation method for washing human PESA sperms. The method comprises the following steps: adding 0.5 mL of a culture medium with a concentration gradient of 80% into a conical pipe, then adding 0.5 mL of a culture medium with a concentration gradient of 40% on the liquid surface to form clear liquid layering; fully mixing an epididymis liquid sample, slowly adding 0.5 mL of epididymis liquid on the liquid surface of the culture medium with a density gradient of 40% to form clear liquid layering; carrying out centrifugation under 400*g for 15 minutes to precipitate high quality sperms in the bottom of the conical pipe; carefully absorbing the sperms out of the conical pipe, suspending the sperms on an HET culture medium with a volume of 5 mL, carrying out centrifugation under 300*g for 8 minutes to remove the residual density gradient culture medium; absorbing the sperms out, suspending the sperms on a sperm capacitation culture medium, carrying out centrifugation under 300*g for 5 minutes to remove the residual HTF culture medium; removing the supernate, fully mixing the capacitation liquid with a volume around 0.5 mL with the sperms, and finally storing the mixture in an incubator with a temperature of 37 DEG C and a CO2 concentration of 5% for later use.

The invention belongs to the biotechnology field and discloses a use of an epididymis specific expression gene HongrES1 or an agonist thereof or an antagonist in the preparation of compositions for regulating sperm capacitation of mammals; the invention further discloses a method for screening effective substances for regulating the sperm capacitation of the mammals by taking the epididymis specific expression gene as a target point. The invention firstly reveals that the HongrES1 has an important role on the aspect of regulating the sperm capacitation and the fertility of the mammals, thereby providing the effective target point for researching male reproduction.

The invention relates to a non-animal origin tree shrew sperm in vitro capacitation liquid and application thereof, belonging to the technical field of animal reproductive biology. The specific stepsare as follows: preparing tree shrew ishing semen and capacitation liquid; separating and removing the epididymis cauda of tree shrews; placing the epididymis cauda in centrifugal tubes containing ished semen and incubating for 10 min to allow sperm to swim out. transferring the sperm suspension to the sterilized centrifuge tube, adding the semen ishing liquid, gently mixing, and centrifuging; discarding supernatant, adding semen ishing liquid, and centrifuging; discardingthe supernatant and resuspending the sperm in the capacitation solution to make the sperm concentration 10* 106 sperm / mL.5 ml of that semen is divided into incubation tubes and incubating in a CO2 incubator. A method for in vitro capacitation of tree shrew spermatozoa adopting animal-free component capacitation liquid avoids that risk of introduce pathogenic factors into in vitro capacitation or in vitro fertilization system, and is easy to carry out quality control on the capacitation liquid.

The invention discloses a quality detection method of experimental primate animal sperms and provides a quality detection kit of primate animal sperms. The quality detection kit of primate animal sperms comprises a sperm capacitation solution. A method for preparing the sperm capacitation solution comprises the step of mixing NaCl, KCl, MgCl2, CaCl2, NaH2PO4, NaHCO3, glucose, sodium pyruvate, sodium lactate and Hepes with water to obtain the sperm capacitation solution, wherein the concentration of NaCl is 114mM; the concentration of Cl is 3.2mM; the concentration of MgCl2 is 0.5mM; the concentration of CaCl2 is 2.0mM; the concentration of NaH2PO4 is 0.4mM; the concentration of NaHCO3 is 24.9mM; the concentration of the glucose is 5.6mM; the concentration of sodium pyruvate is 0.5mM; the concentration of sodium lactate is 10mM; the concentration of Hepes is 10mM. By virtue of the method, the damage to the reproductive system caused by collecting seminal fluid through electrical stimulation can be avoided; meanwhile, the method is capable of quickly detecting the quality of the sperms while sampling experimental animals, so that the method has a broad application prospect in research such as food safety evaluation.

The invention provides a method for obtaining a sterile mammal with rapidness and high efficiency. The method comprises the following steps: a) obtaining a sperm of a male mammal, and subjecting the obtained sperm to capacitation; b) obtaining an ovum of a female mammal; c) preparing an oosperm by utilizing the sperm obtained in the step a) and the ovum obtained in the step b); d) obtaining an aspermous male mammal; e) preparing a pseudopregnant female mammal; f) preparing an embryo by utilizing the oosperm obtained in the step c); g) obtaining a pseudopregnant female mammal containing the embryo obtained in the step f); and h) subjecting the female mammal obtained in the step g) to sterile culture so as to obtain the sterile mammal. The method provided by the invention has the advantages of short time, fast action, high efficiency and small breeding population, so the sterile mammal used for various purposes can be provided with rapidness and high efficiency.

The invention provides a reagent kit for quality evaluation of sperm after in vitro capacitation and a use method of the reagent kit. The reagent kit is stuffed with a detection buffer solution 1, a BWW culture solution, an in vitro capacitation solution, a fluorescent reagent, a sialidase standard stock solution and a detection buffer solution 2, wherein the detection buffer solution 1 is a phosphate buffer solution or the BWW culture solution; the detection buffer solution 2 is the phosphate buffer solution containing 0.05mmol / l sodium acetate and 0.25<mu>g / <mu>l bovine serum albumin. Through in vitro capacitation of the sperm, the activity of sialidase in a capacitation solution can be detected, so that a diagnosing basis and clinical consultation can be provided for patients suffering from male sterility of unknown clinical aetiology.

The invention provides a detection kit of phosphorylation of sperm tyrosine. The detection kit comprises the following components: a capacitation buffer liquid, a fluorescent combined object and a fluorescent solid sealing object. The invention also provides a method for detecting the phosphorylation of the sperm tyrosine by using the kit. Compared with the prior art, according to the invention, each link of methodology is optimally designed, which includes the best buffer liquid matching of capacitation and an upstream method, a stable reaction place offering by special tissue carrier sheets, sperm double-labeling technique formulation combined with specificity, and influences to the detection result by non-sperm substances of cast-off cells in a seminal fluid are eliminated, thereby guaranteeing the methodology reliability, detection result specificity and good repeatability. Because an optimized detection scheme is adopted, the operation steps and the operation time are reduced, and the detection efficiency is improved greatly; because the optimized detection scheme is adopted, the reagent structure is simplified, and the detection cost is lowered to a greater degree; and because the operation is simple, ready-to-use available reagents are adopted, and the performance is stable.

The invention discloses a bacteriostatic uterine horn insemination diluent suitable for donkey semen, and also discloses a preparation method of the antibacterial uterine horn insemination diluent suitable for donkey semen. The advantages are: by adding an appropriate amount of taurine, hypotaurine, bovine serum albumin, heparin sodium and progesterone, it can promote sperm capacitation in vitro and maintain sperm motility. It can greatly improve the utilization rate of male donkey semen, and further increase the number of fertilized female donkeys, so that more female donkeys can be conceived and promote donkey breeding. The development and expansion of the enterprise's breeding scale will improve the economic benefits of the enterprise; by adding an appropriate amount of cefoxitin sodium, amphotericin and polymyxin, it can effectively reduce the probability of metritis in female donkeys, and ensure a healthy uterine environment for female donkeys. Promoting the application and promotion of deep angle insemination can provide a strong guarantee for improving the conception rate.

The invention discloses an HTF liquid drop for improving fertilization ability of mouse sperms and a preparation method and a use method of the HTF liquid drop. The preparation method comprises the following steps that 200 [mu]L / drop HTF liquid drop is prepared in a culture dish, tissue paraffin oil covers the HTF liquid drop, and the HTF liquid drop is put in an incubator for balancing; the mouse SERPINA5 artificially synthesized protein powder is prepared into a mouse SERPINA5 artificially synthesized protein concentrated storage by using serum-free HTF, and the concentration of the mouse SERPINA5 artificially synthesized protein concentrated storage is 240-260 [mu]g / mL; and the mouse SERPINA5 artificially synthesized protein concentrated storage is added into the HTF liquid drop, so that the concentration of the mouse SERPINA5 artificially synthesized protein in the HTF liquid drop is 1-2 [mu]g / mL. According to the HTF liquid drop for improving the fertilization capacity of the mouse sperms and the preparation method and use method of the HTF liquid drop, the in-vitro fertilization capacity of the mouse sperms can be effectively improved, and a reference method is provided for optimizing the sperm quality in an ART sperm in-vitro capacitation process.

The invention provides a method for evaluating the quality of sperm obtaining energy in vitro. The method specifically includes the following steps of A, collecting a fresh liquefied sperm specimen, washing sperm with a phosphate buffer or a BWW culture solution, sufficiently removing residual seminal plasma components, and separating out sperm in a centrifugal mode; B, adding the BWW culture solution to sperm separated out in the step A, obtaining upstream sperm through an upstream method, counting the number of sperm, incubating sperm for 3-4 hours in a CO2 cell culture box with 5% of in-vitro energy obtaining liquid, conducting centrifugal separation on the obtained supernate, and collecting supernate as a to-be-tested sample; C, measuring the activity of neuraminidase in sperm energy obtaining liquid, and finally obtaining the value of neuraminidase activity / sperm number to evaluate the quality of sperm. By detecting the activity of neuraminidase in sperm in-vitro energy obtaining liquid, diagnosis basis and clinical queries are provided for male infertility patients of unknown clinical reasons.

The invention belongs to a transgenic field, and relates to a treatment method for preparing sperm of a transgenic mouse. The treatment method for preparing sperm of the transgenic mouse, comprises: incubating mouse epididymal sperm at 37 DEG C in a 5% CO2 incubator for 8-10min, mixing a linear DNA fragment containing a target gene into the sperm, ice-bathing the sperm at 2-5 DEG C for 25-30min, heat-shocking the sperm at 40-45 DEG C for 1-3min, ice-bathing the sperm at 2-5 DEG C for 3-5min, and then incubating the sperm at 37 DEG C in a 5% CO2 incubator for 50-70min for capacitation The method can raise permeability of a sperm membrane, is convenient for introducing a gene fragment into the sperm, and raises transgenic efficiency. The invention solves the problems of low efficiency, bad stability and difficult passage when sperm is used to prepare the transgenic mouse. An effective method for preparing the transgenic mouse is disclosed.

The invention relates to a new method for cloning embryo with bovine sperms, which comprises the steps as follows: (1) a collected bovine ovary is cleaned, and an ovarium mound-oocyte complex is drawn out for in vitro culture to lead the oocyte to be cultured and developed to a second meiosis metaphase; (2) then dyeing and fluorescence localization are carried out on the nucleolus of the oocyte,and the nucleolus is denucleated to obtain the denucleated bovine oocyte; (3) the semen of an excellent breeding bull is collected or the commercial semen of a self-sex controlled excellent breeding bull is selected and stored in a liquid nitrogen container; (4) the semen is unfrozen and capacitation treatment is carried out on the semen; then the semen is arranged in the injecta of the oocyte; (5) two capacitation sperms are injected into the cytoplast of the denucleated bovine oocyte to obtain a sperm reconstructed embryo; (6) the sperm reconstructed embryo is activated and cultured. The invention overcomes the single mode that the donor cell in the traditional nuclear transfer can only be a somatocyte, and is a new method for generating a bovine embryo by using double sperms to replacethe somatocyte as a donor to be injected into the cytoplast of denucleated bovine oocyte.

The invention provides an RSSI-based low-power consumption passive wireless node networking method. According to the invention, networking is carried out by use of self-capacitation passive wireless nodes, and an optimal transmission router and an Internet with smallest communication consumption are established, so stability and uniformity of the network are ensured and an effect of low-power consumption high-quality wireless communication of a wireless network is achieved on a bidirectional communication function.

A buffer solution for sperm capacitation is composed of a buffer solution comprising an active ingredient astaxanthin combined with serum albumin and a phosphate buffer solution, and the phosphate buffer solution is composed of monopotassium phosphate KH2PO4 buffered by dipotassium phosphate K2HPO4.

The present invention relates to a method of predicting animal litter size using a fertility-related protein marker, and more particularly, to the discovery of a sperm marker that is expressed differently depending on animal fertility, a marker composition for predicting litter size, which comprises an antibody that binds specifically to the marker, and a method of predicting animal litter size using the marker composition. Moreover, the present invention relates to a method of predicting animal semen quality and litter size by chlortetracycline staining, and more particularly, to a method of predicting of animal litter size by measuring the motility, motion kinematics or capacitation status of sperm. When the animal sperm-derived protein marker according to the present invention is used, the litter size of individuals can be predicted by analyzing a protein that is expressed differently depending on litter size. When the method of animal semen quality and litter size by chlortetracycline staining is used, the fertility of sperm and the litter size of individuals can be predicted. According to the present invention, superior species having high sperm fertility and high litter size can be selected based on information provided by the method. Thus, the present invention is highly useful for the sustainable production of animals.

The invention belongs to a transgenic field, and relates to a treatment method for preparing sperm of a transgenic mouse. The treatment method for preparing sperm of the transgenic mouse, comprises: incubating mouse epididymal sperm at 37 DEG C in a 5% CO2 incubator for 8-10min, mixing a linear DNA fragment containing a target gene into the sperm, ice-bathing the sperm at 2-5 DEG C for 25-30min, heat-shocking the sperm at 40-45 DEG C for 1-3min, ice-bathing the sperm at 2-5 DEG C for 3-5min, and then incubating the sperm at 37 DEG C in a 5% CO2 incubator for 50-70min for capacitation The method can raise permeability of a sperm membrane, is convenient for introducing a gene fragment into the sperm, and raises transgenic efficiency. The invention solves the problems of low efficiency, bad stability and difficult passage when sperm is used to prepare the transgenic mouse. An effective method for preparing the transgenic mouse is disclosed.