31 results about "Second polar body" patented technology

Disclosed is a method for using a first polar body and a second polar body as well as a single-cell embryo to carry out whole-genome non-exponential amplification and high-throughput genome sequencing, so as to perform preimplantation genetic diagnosis for genetic disease and testing for pathogenic genes causing repeated miscarriages. The method of the present invention comprises the following steps: (1) obtaining oocytes and embryos, and carrying out separation and genome amplification of first and second polar bodies and single-cell embryos; (2) establishing a genome sequencing library and sequencing, and carrying out bioinformatic analysis of the genome to obtain a gene spectrum and information concerning the number of copies of chromosomes and fragments thereof; (3) determining the chromosome ploidy of the polar bodies and embryos as well as information concerning defects, replication and point mutation in the chromosome fragments; (4) selecting normal or suitable embryos for implantation.

The invention relates to a human parthenogenetic embryo stem cell line with two active X chromosomes and derivatives thereof. A karyotype of the human parthenogenetic embryo stem cell line is 46, XX before 10 generations; the karyotype of the cell line becomes a chimera from the 20th generation; all cells of the cell line lose an active X chromosome at the 35th generation; and in the inactivation detection of the X chromosome, only the state of the active X chromosome is maintained all along, but the presence of the inactivated X chromosome is not discovered. The parthenogenetic activation can be achieved by adopting an artificial activation condition causing a second polar body to be discharged; therefore, the human parthenogenetic embryo stem cell line and the derivatives thereof can be obtained. The cell line and the derivatives thereof have important significance on researching the characteristics of the cell on genetics and epigenetics and further applying the cell to clinically treating certain diseases; and a cell source for cell replacement therapy can be obtained by transforming genes of the cell or modifying the genes on the epigenetics.

The invention discloses a production method of triploid pseudobagrus fulvidraco, which belongs to the fish culture technology field. Artificial insemination is performed to the pseudobagrus fulvidraco female parent and male parent which are obtained through artificial induced spawning, after 10 to 15 minutes, triploid pseudobagrus fulvidraco is obtained through induction, and the duration time for induction is 10 to 20 minutes. After the triploid pseudobagrus fulvidraco is obtained through induction, the multiple of the chromosome is measured through the technologies, such as the karyotype analysis of chromosome, the flow cytometer, etc., and the triploid pseudobagrus fulvidraco is identified and sieved for culture. The method restrains the releasing time of the second polar body through time control on the artificial insemination, the induction through the physical or chemical method, and control on the induction duration time, and thereby the inductivity and the survival rate of the triploid pseudobagrus fulvidraco are greatly increased, and according to the experiment statistics, the artificial inductivity of the achieved triploid pseudobagrus fulvidraco is more than 50 percent, and the survival rate is more than 80 percent.

The invention provides a method for culturing a Crassostrea angulata tetraploid. The method comprises the steps of firstly culturing a pacific oyster tetraploid, hybridizing the pacific oyster tetraploid with Crassostrea angulata so as to culture a hybridized triploid, fertilizing the hybridized Crassostrea angulata triploid with a Crassostrea angulata diploid sperm, carrying out anesthetization by virtue of magnesium chloride mixed liquid to inhibit the release of a second polar body so as to generate the tetraploid, wherein an inductive agent is safe and toxic-free. According to the method,the long induction process of the Crassostrea angulata triploid is solved, the survivable and stable tetraploid Crassostrea angulata can be cultured by virtue of the technical routes of hybridization,high permeation, anesthetization, molecular marking and the like, and the foundation is laid for the large-scale seedling growing of triploid Crassostrea angulata.

The invention discloses a gynogenesis method of grass carp and application thereof. The method comprises the following steps: taking the diploid sperms of a properly inactivated allotetraploid cruciancarp as a stimulus to stimulate mature eggs of the grass carp, after activating for 2-3 min or fertilizing for 20-22 min, carrying out the water-cooling shock treatment for 10-12 min at 4-6 DEG C, and hatching and controlling the temperature by to obtain the gynogenesis grass carp restraining the discharge of a second polar body or the first cleavage. The method greatly improves the hatching rateand the survival rate of the gynogenesis diploid grass carp, simplifies the preparation process, and can prepare a gynogenesis grass carp colony with good characters and stable heredity.

The invention discloses a preparation method of young mollusks of Hongkong tetraploid oysters. The method comprises the following technical steps of selectively choosing triploid eggs, carefully choosing diploid sperms, inhibiting second polar bodies of oosperms, obtaining tetraploid young mollusks, and raising tetraploid progenies. The feature of normal reduction mitosis of some triploid eggs is adopted so as to create a raising method for preparing tetraploid oysters by inhibiting second polar bodies of oosperms (triploid eggs +diploid sperms). The method differs from a method for obtaining tetraploid oysters by inhibiting first polar bodies of oosperms. The preparation method makes a great difference in a traditional preparation mode and helps break the monopolies of the US, France, Korea and other foreign countries on raisins technology of tetraploid oysters. It is the first time to obtain livable young mollusks of Hongkong tetraploid oysters. The method makes the biological method of producing 100% Hongkong triploid oysters feasible, promote the steady development of the industry of Hongkong oysters in the coastal region of southern China and offers the possibility of achieving improved varieties of the industry. The preparation method of young mollusks of Hongkong tetraploid oysters is being advantaged by being implementable, reusable, and applicable.

The invention discloses a method for inducing triploid of Haliotis discus Hannai through medicament, which comprises the following steps, preparing 6-dimethoxy aminopurine solution, obtaining abalone sperms and ova through the conventional method, obtaining oosperm through artificial insemination, placing oosperm of seed abalone into sea water, finally inhibiting the second polar body of the Haliotis discus Hannai with 6-dimethoxy aminopurine solution.

The invention provides a cultivation method for carassius auratus gynogenesis fries. The method comprises the steps of selecting sexually mature diploid carassius auratus parents and collecting sperm and ovums; performing ultraviolet irradiation on Hanks liquid-diluted sperms to inactivate genetic materials thereof; performing artificial insemination on the inactivated sperms and the ovums; restraining the discharge of second polar bodies of meiosis by using a cold shock method to double genomes of the ovums and thus to cultivate viable carassius auratus gynogenesis diploid fries. Microsatellite marker identification proves that the genetic materials of all the gynogenesis fries are from the female parent; morphological measurement indicates part of the gynogenesis fries has obvious early growth dominance. The method has the advantages of simple operation, high efficiency, stability and high practicality.

The invention relates to a method for inducing gynogenesis of a Siberian sturgeon by using an amur sturgeon sperm, which comprises the following steps: (1) inactivating a genetic material of the amur sturgeon sperm; (2) acquiring Siberian sturgeon unfertilized ova and performing insemination with an inactivated sperm of an amur sturgeon; (3) inducing the gynogenesis of the Siberian sturgeon through heat shock: inducing the gynogenesis of the Siberian sturgeon by adopting a method for inhibiting a second polar body from being discharged through the heat shock , wherein the starting time of the heat shock is 0.3 tau0 (18 minutes at a temperature of below 16 DEG C) after fecundation, the duration of the heat shock is 2 minutes, and the temperature of the heat shock is 37 DEG C; and (4) cultivating a gynogenesis induced egg. A super-female fish of the Siberian sturgeon cultivated by the technique of the method can produce gynoecy fish fries by mating with a normally cultivated milter, thus the proportion of female individuals in cultured Siberian sturgeon population is improved, and the cultivation benefit is improved.

The invention discloses an inducing method of a Nibea albiflora gynogenesis diploid. The technical scheme of the invention is that the inducing method comprises the steps of genetic inactivation of sperms of Nibea albiflora and diploidzation of ova of the Nibea albiflora; the inducing method is mainly characterized by firstly enabling the sperms of the Nibea albiflora to be subjected to the genetic inactivation by using ultraviolet irradiation, then activating the sperms and the ova by using seawater after the sperms are mixed with the ova of the Nibea albiflora, starting gynogenesis of the ova of the Nibea albiflora, restraining the discharge of a second polar body by using cold shock, doubling genomes of the ova, and inducing the Nibea albiflora gynogenesis diploid. The inducing method disclosed by the invention has the advantages that the operation is simple, the needed cost is lower, the practicability is stronger, and the like.

The invention relates to a physical method of polyploidy-breeding by introducing allotriploid scallops, put female scallops in filter seawater at the temperature of 16íµ after being dried in shade for 40 minutes to stimulate ovulation; put male scallops in filter seawater at the temperature of 16íµ after being dried in shade for 1 hour to stimulate sperm-drain, and then put said ovums and sperms produced in filter seawater at the temperature of 20íµ for fertilization following the mixing rate of 1í†5. When 30% of ovums have been drained out of second polar bodies after fertilization, force pressure of 20-30 MP for 20-10 minutes, hatch fertilized ovums according to the conventional method and get triploid scallops. The scallops cultured in said invention are characterized on fast,growth, high infertility and fresh texture, could make up for the shortages of low production and induced texture during the multiplication periods caused by diseases and high-density culturing in recent years.

The invention discloses a method for obtaining takifugu rubripes androgenesis haploids. The steps of the method are as follows: (a) the spawning of male takifugu rubripes and female takifugu rubripes with well-developed gonads is induced, the sperms of the male takifugu rubripes and the ova of the female takifugu rubripes are respectively taken, and the sperms and the ova are mixed to form fertilized ova; (b) the androgenesis of the takifugu rubripes is induced by cold shock. The method adopts the sperms of the male takifugu rubripes and the ova of the female takifugu rubripes to form the fertilized ova; cold shock is used for processing (30 to 60 minutes of immersion in ice water of 2 DEG C to 4 DEG C 5 to 8 minutes after fertilization), the egg nuclei and second polar bodies of the takifugu rubripes are induced to release together, and thereby fertilized ova which only have sperm nuclei are formed. Because the genetic inactivation of ova is not required, a complex operation procedure is avoided, the method is cheap and easy to operate, the problem that conventional physiochemical induction can injure fertilized ova is solved, the androgenesis rate is increased, and the method is suitable for use in mass production.

A method for inducing the polyploid of marine shellfish includes such steps as measuring the salinity of culturing seawater, taking the sperms and ova of shellfish, artificial insemination, adding fresh water to seawater until its salinity is 4-16 when 40-50% of fertilized ova is releasing the first polar bodies, stirring for 10-15 min for suppressing the release of the second polar bodies, and incubating the fertilized ova in normal seawater.

The invention relates to chromosome operation technology, and particularly relates to a continuous batch induction method for salmo salar triploids. The method includes the following steps: collectingsalmo salar sperm eggs to carry out artificial insemination, and dividing fertilized eggs into 2-3 groups; and carrying out a hydrostatic pressure induction treatment on each group in a successive and continuous mode to inhibit emissions of second polar bodies, thereby obtaining the salmo salar triploids. According to the hydrostatic pressure induction treatment, 300-400 DEG C x minute (temperature x time) after insemination, the fertilized eggs in Group 1 is subjected to the hydrostatic pressure induction treatment to obtain the triploid salmo salar; after the induction, the fertilized eggsin the Group 1 are taken out, and then the fertilized eggs in Group 2 are immediately performed with the hydrostatic pressure induction according to the above-mentioned process, thus the same batch offertilized eggs can be induced and treated continuously. According to the invention, the method can be used for continuous and efficient induction to obtain 100% salmo salar triploids.

The invention relates to a method for in-vitro fertilization by taking testicular sertoli cells as trophoblasts. The method includes the steps of (1), subjecting the testicular sertoli cells to isolated culture to prepare the sertoli cell trophoblasts; (2), guiding sperms into capacitation liquid containing well balanced HTF for incubation for 1 hour; (3), collecting ova; (4), transferring washed ova into a balanced culture medium with a testicular sertoli cell monolayer; (5), adding 20 microliters of incubated sperm suspension liquid into a culture solution containing the ova; (6), placing a culture dish back to an incubator with 5% CO2 at the temperature of 37 DEG C for incubation, and holding for 5-6 hours; (7), picking out the ova with second polar bodies and pronuclei and transferring into the culture medium for culturing. The method for in-vitro fertilization by taking the testicular sertoli cells as the trophoblasts has the advantages of high insemination rate and developmental rate.

A culture method for inducing triploid Hyriopsis cvmingii Lea by using 6-dimethylamino purine includes such steps as obtaining the ova of triploid Hyriopsis cvmingii Lea and fertilizing with sperm from diploid, using an inhibitor to suppress a first or a second polar body, which is characterized in that the inhibitor is 6-dimethylamino purine for short 6-DMAP. The invention also discloses a methof for culturing pearl by using the triploid Hyriopsis cvmingii Lea. The inhibitor of the invention has characteristics of low cost and toxicity, and adapts to culturing pearl in large water area.

The invention relates to a production method of triploid shellfishes, in particular to a production method of the triploid shellfishes from diploid shellfish production to tetraploid mother shellfish production to triploid shellfish production. The production method is characterized by comprising the following steps of S1, a first stage of fertilization of diploid female eggs and diploid male sperms; S2, a second stage in which the release of a first polar body or a second polar body of fertilized eggs is prevented in the first stage S1 and triploid females are produced; S3, a third stage in which the female eggs in the second stage S2 are fertilized with the diploid male sperms; S4, a fourth stage in which the release of the first polar body is prevented from occurring from the eggs of the third stage S3 and tetraploid males are produced; and S5, the fifth stage in which the diploid female eggs are fertilized with the male sperms in the fourth stage S4, and the triploid shellfishes are generated.