47 results about "Bovine embryo" patented technology

This disclosure provides ungulate embryonic stem cells (ESCs) derived from the inner cell mass of pre-implantation blastocysts or pluripotent cells from embryos. From an agricultural and biomedical perspectives, the derivation of stable ESCs from domestic ungulates is important for genomic testing and selection, genetic engineering, and providing an experimental tool for studying human diseases. Cattle are one of the most important domestic ungulates that are commonly used for food and bioreactors.

The invention provides a bovine embryo vitrification tube swinging thawing and direct transplanting method and belongs to the technical field of embryo low-temperature biology. The method includes (1) freezing a bovine embryo and using a fine tube to first add a thawing liquid followed by air; (2) loading a freezing liquid followed by air; (3) loading a freezing liquid, placing the bovine embryo in the freezing liquid, and then loading air; (4) loading a thawing liquid, closing a tube opening, and performing freezing; (5) performing tube swinging thawing and direct transplanting, wherein the freezing liquid is an mPBS solution containing ethylene glycol (EG), polysucrose and sucrose, and the thawing liquid is an mPBS solution containing fetal bovine serum (FBS). According to the bovine embryo vitrification tube swinging thawing and direct transplanting method, the operation is simple and convenient, requirements for skills of operators are not high, the frozen bovine embryo can be directly transplanted after tube swinging thawing, the thawed embryo has a survival rate of 100% and an expansion rate of 94%, a pregnancy rate reaches 57% after transplantation, and the method is efficient, simple, easy to operate, and applicable to industrialized popularization.

The invention discloses a fertilization solution for in-vitro fertilization of cattle and a use method thereof. The fertilization solution includes a SOF basal medium without inositol, and also includes the following additives: L-arginine, L-aspartic acid, L-carnitine, adrenaline, penicillamine, hypotaurine, and heparin. The fertilization solution of the invention can effectively ensure the in-vitro fertilization rate of the bovine embryo and the development quality of the fertilized embryo.

The invention relates to the technical field of animal infectious disease prevention and treatment. The invention provides a mycoplasma bovis Mbov0701 mutant gene as well as a mutant strain and application of the mycoplasma bovis Mbov0701 mutant gene. The gene mutant strain has insertion mutation behind a 701 nucleotide site of the Mbov0701 gene, shows a significant growth defect phenotype when co-cultured with bovine embryo lung cells, and shows a small colony form on a PPLO solid culture medium. The Mbov0701 gene encoding protein is exonuclease MbovP701. The gene mutant strain can be applied to the fields of mycoplasma bovis metabolism physiology, pathopoiesis, prevention and treatment medicine preparation and the like.

The invention discloses a novel bovine embryo transplantation gun capable of bending and extending forward along with the uterine horn, and relates to the field of embryo transplantation guns, the novel bovine embryo transplantation gun comprises a gun barrel, a gun core and a culture solution thin tube, the culture solution thin tube is installed in a barrel hole in the inner end of the gun barrel in a matched mode, a piston is arranged in the culture solution thin tube, and the gun core is arranged in the gun barrel. According to the invention, an embryo in the slim tube is pushed into the extension catheter through the piston, and then the piston is punctured through the hollow steel needle to be communicated with the inner cavity of the slim tube; an operator pushes a piston handle of the injector to enable gas in the injector to enter the extension catheter through the hollow steel needle, and an embryo in the extension catheter is driven into the uterine horn of a cattle by the gas, so that the problem that the embryo cannot be driven into the uterus by a gun core after a slim tube is connected with the extension catheter is solved, and the operator does not need to grab the front end of the uterine horn; the operation difficulty is greatly reduced, the use of operators is facilitated, and the conception rate is indirectly improved.

Arrays of nucleic acid molecules, kits, methods of genotyping and marker assisted bovine breeding methods based on novel SNPs on genes of the bovine transforming growth factor-β (TGF-β) signaling pathway for improved bovine fertilization rate. The methods and compositions of the present invention are related to SNPs in the DNA-binding protein inhibitor 3 (ID3) gene, and in the bone morphogenetic protein 4 (BMP4) gene corresponding to position 2702 of SEQ ID NO: 2. Also disclosed are methods for determining viability of developing bovine embryos by measuring the expression level of one or more target genes in the TGF-signaling pathway, and selecting for implantation only embryos whose target gene expression level is not up-regulated.

The invention provides embryo antioxidation related lncRNA and application thereof, and belongs to the technical field of lncRNA function research. A nucleotide sequence of the lncRNA CUFF.97956.1 is as shown in SEQ ID No.1; a target gene of the lncRNA CUFF.97956.1 is a PANX1 gene; the expression quantity of the lncRNA CUFF.97956.1 in the bovine embryo is remarkably reduced after glutathione treatment; the lncRNA CUFF.97956.1 and the target gene thereof are related to the antioxidant and developmental capacity of the embryo; and the lncRNA CUFF.97956.1 and the target gene thereof provide technical support for improving the developmental potential of the in-vitro embryo.

The invention relates to a method for gender identification of early bovine embryos, comprising the following steps: performing in vitro fertilization of oocytes and sperm to obtain morulae; cutting the morulae in the segmentation liquid droplet with a segmentation knife to obtain embryonic tissues containing 10 to 12 cells As a sample for gender identification; the ring-mediated isothermal amplification method gender identification kit makes samples and prepares Master Mix; after the samples are mixed, put the reaction tube in the ring-mediated isothermal amplification real-time turbidity detection system for reaction, and determine the sample according to the results One of the three cases needs to be redone for male, female, uncertain. The method of the invention presents excellent technical effects as described in the specification.

The present invention relates to a convenient sex determination method of bovine embryos, the identification rate of the blood genome achieves 100%, and the identification rate of the embryo sex exceeds 99.0%. This method has convenient operation, high sensitivity, good practicability and simple equipment, and can determine the embryo sex in 1 hour. The formed detection kit of embryo sex can be applied to production absolutely and the cost is only 1 / 10 of products of the same kind. In the present invention 6 specific primers are designed according to the bovine specific sequences and male specific sequences, and 6 specific sites can be recognized. Large amount of DNA fragments of different sizes are amplified in a short time using a dumbbell-shaped structure unit as a core that is formed by the target sequences. The only requirement is aqueous thermostat at 60-65DEG C. dNTPs precipitation is used in the determination of the result (The negative result is that a white band appears in the middle part of the reaction tube).

The invention discloses a bovine embryo sex development related gene SOX9, relating to the biological field. A nucleotide sequence of the related gene is as shown by SEQ ID NO.1. The invention also provides an encoding protein of the gene, and an amino acid sequence of the encoding protein is as shown by SEQ ID NO.3. The invention also provides an extracting method of the gene, comprising the following steps of: extracting a total RNA (Ribonucleic Acid) from a bovine embryo genital ridge, amplifying by an RT-PCR (Reverse Transcription-Polymerase Chain Reaction) method, and cloning to a complete expressed sequence of the bovine embryo sex development related gene SOX9 by combining an RACE and nest-type PCR technology. The gene can be used for controlling the sex of a bovine offspring and has good application prospect.

The invention provides an assembly for cervical dilatation before cattle embryo transfer. The assembly comprises a jacket body with an inner cavity, a support frame assembly is arranged in the jacket body and provided with a dilatation assembly capable of swelling by absorbing water and can be pushed by an ejector rod to drive the dilatation assembly to move in the axial direction of the jacket body, and the jacket body is provided with a control assembly which can control the support frame assembly and the jacket body to be in a separable state or an inseparable state. According to the assembly, an embryo can more easily enter the uterus through the cervix uterus during embryo transfer, therefore, the success rate of embryo transfer is increased, the reproductive system infection rate caused by embryo transfer is decreased, stress responses of cattle are reduced, and the convenient operation advantage is achieved.

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 relates to a method for bovine in-vitro fertilized embryo culture and a used culture solution. Particularly, on one hand, the invention relates to a method for culturing a bovine in-vitro fertilized embryo. The method comprises the following steps of putting the bovine in-vitro fertilized embryo into a bovine in-vitro fertilized embryo culture solution and carrying out embryo in-vitro culture under the conditions that the temperature is 38.5 DEG C, the content of CO2 is 0.5% and the humidity is 100%. The invention further relates to a bovine in-vitro fertilized embryo culture solution. The culture solution comprises NaCl, KCl, NaHCO3, MgCl2, KH2PO3, sodium pyruvate, glucose, calcium galactonate, fetal calf serum, L-glutamine, an essential amino-acid, a nonessential amino acid, glutathione and water. The method and the used culture solution have excellent technical effects in the specification, for example, the method has a higher blastocyst hatching rate when the bovine in-vitro fertilized embryo is cultured.