39results about How to "Increase the cost of feeding" patented technology

The invention discloses a CRISPR (clustered regularly interspaced short palindromic repeats)/Cas9 system and application thereof to construction of swine-derived recombinant cells with insulin receptor substrate gene defects. The invention provides an sgRNA (ribonucleic acid) combination which is composed of sgRNAIRS1-1, sgRNAIRS1-3, sgRNAIRS2-2 and sgRNAIRS2-3. The invention provides a plasmid combination. The plasmid combination is composed of four plasmids, and the four plasmids are respectively subjected to transcription to obtain sgRNAIRS1-1, sgRNAIRS1-3, sgRNAIRS2-2 and sgRNAIRS2-3. Theapplication of the sgRNA combination or the plasmid combination is as follows: preparation of recombinant cells; preparation of a diabetic cell model; preparation of a diabetic animal model. Accordingto the invention, sgRNAIRS1-1 is as shown in SEQ ID NO:8; sgRNAIRS1-3 is as shown in SEQ ID NO:10; sgRNAIRS2-2 is shown as SEQ ID NO:14; and sgRNAIRS2-3 is as shown in SEQ ID NO:15. A solid foundation is laid for the preparation of diabetic pig models, and important application value for the research and development of diabetic medicaments is achieved.

The invention discloses a Chinese herbal medicament pigeon feed, which is prepared from the following raw materials in percentage by weight: 37 to 46 percent of corn, 18 to 27 percent of pea, 7 to 11 percent of bean cake, 2 to 4 percent of fish meal, 1 to 5 percent of fructus cannabis, 8 to 12 percent of laterite, 3 to 5 percent of bentonite, 9 to 13 percent of yellow sand, 2 to 4 percent of common salt, 0.5 to 1 percent of binding agent, and 2.4 to 3.2 percent of Chinese herbal medicament mixture containing pilose asiabell root. The Chinese herbal medicament mixture containing the pilose asiabell root is prepared by mixing the pilose asiabell root, astragalus, gentian, liquorice root, common andrographis herb and plantain herb according to the weight ratio of 3:2:1:1:0.5:0.5. For further improvements, the feed also comprises 0.3 to 0.5 percent of Chinese caterpillar fungus and 0.5 to 0.8 percent of lucid ganoderma. The Chinese herbal medicament pigeon feed has the characteristics of good palatability, comprehensive nutrition, obvious health-care and disease prevention functions and low cost.

The invention discloses a method for raising geese in a corn field. The method comprises the steps that firstly, corn is planted and goslings are prepared; the goslings are raised in the corn horn mouth stage at the end of June and certain feed is offered everyday in the stage; the goslings are raised in the corn field in the corn booting stage or the corn heading stage in the middle ten days of July; certain feed is offered so that growth of the goslings can be maintained and the feed is offered at night; geese are obtained in the corn milk ripe stage or the corn dough stage almost at the beginning of October and the corn is harvested ten days after the geese are obtained. According to the method for raising the geese in the corn field, corn cultivation and goose raising are organically combined and the cost benefits of the geese and life benefits of the corn are improved; the geese are obtained when the corn is mature, the goose raising time is prolonged, the difficulty of technology conduction is reduced, the yield of the corn is guaranteed, and the yield of the geese is increased as well. In addition, the production benefits, the land use efficiency, the light energy use efficiency, the production efficiency and the economic benefits are improved and the ecological benefits are good.

The invention discloses an ecological breeding oxygenation device for soft-shelled turtle. The device comprises a floating boat, a fixing plate, a motor, a sealing cylinder, a coupling, a hollow tube, a rotating exhaust tube, an exhaust fan and an air tube. Compared with the prior art, the device has the advantages that air is pumped into the sealing cylinder through the exhaust fan and then is guided into the hollow tube through the sealing cylinder, the motor drives the hollow tube to rotate, then the exhaust tube rotates together to exhaust air into the water, thus the oxygen content in water is increased, and as a result, that the soft-shelled turtle is out of appetite due to oxygen deficit and grows slowly and the breeding cost is increased can be avoided.

The invention discloses a method for constructing high-quality porcine nuclear transplantation donor cells with high lean meat percentage, fast growth and resistant to porcine reproductive and respiratory syndrome and series diarrhea diseases and application of the high-quality porcine nuclear transplantation donor cells. A CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats/CRISPR associated protein 9) system for porcine MSTN-SST-CD163-pAPN four-gene editing contains a Cas9 expression vector and gRNA (guide Ribonucleic Acid) expression vectors aiming at a porcine MSTN gene, an SST gene, a CD163 gene and a pAPN gene, and plasmid complete sequence of the Cas9 expression vector is as shown in SEQ ID NO.2. According to the invention, the corresponding gRNA expression vectors are designed for different targets of MSTN, SST, CD163 and pAPN genes, and gRNA with relatively high editing efficiency and the gRNA expression vector are obtained through screening. Through cooperation with the modified Cas9 efficient expression vector for gene editing, the editing efficiency is remarkably improved as compared with that of an original vector.

The invention discloses a CRISPR/Cas9 system for preparing an LMNA gene mutation dilated cardiomyopathy cloned pig nuclear donor cell and application of the CRISPR/Cas9 system. The invention providesa kit which comprises sgRNALMNA-E7-g1 or plasmid pKG-U6gRNA(LMNA-E7-g1).sgRNALMNA-E7-g1, a target sequence binding region of the sgRNALMNA-E7-g1 or plasmid pKG-U6gRNA(LMNA-E7-g1).sgRNALMNA-E7-g1 is shown as nucleotides from the first to the 20th sites in SEQ ID NO: 6. The plasmid pKG-U6gRNA(LMNA-E7-g1) is transcribed to obtain sgRNALMNA-E7-g1. The invention also claims a method for preparing a recombinant cell. The method comprises the following step: co-transfecting a pig cell with plasmid pKG-U6gRNA(LMNA-E7-g1) and plasmid pKG-GE3 to obtain the recombinant cell. The CRISPR/Cas9 system lays asolid foundation for obtaining a dilated cardiomyopathy pig model by a gene editing means, and has great application value for research and development of dilated cardiomyopathy drugs and revealing of pathogenesis of the disease.

The invention relates to a precise combined feeding method for regulating water quality in aquaculture, and belongs to the technical field of the aquaculture. The precise combined feeding method for regulating the water quality in the aquaculture includes steps: using high and low protein feed in mixing mode during the whole aquaculture process, regulating a carbon-nitrogen ratio in the medium period of the aquaculture, and regulating the water quality in the later period of the aquaculture. By using the precise combined feeding method for regulating the water quality in the aquaculture, the water quality can be regulated well, bacilli and lactic acid bacteria in a water body become dominant bacteria, fishes can grow well and quickly by throwing the feed in the later period of the aquaculture, a culture cycle is shortened, yields and efficiency are increased, and a feed coefficient is reduced.

The invention discloses feed for stable breeding of spotted deer in the south. The feed is prepared from the following components in quality ratio: corns, bean meal, wheat bran, rice bran, alfalfa, pine needle powder, kudzu vines, houttuynia cordata, bunge pricklyash leaves, corn straw, pennisetum purpureum schum, carrots, sweet potatoes, vitamin complex, active ferment, amino acid additives and trace element additives. Basal feed, Chinese herbal medicine raw materials and additives such as mineral substances and vitamins are scientifically blended, thereby effectively meeting the energy and nutrition needs of the spotted deer in the growth process, improving the utilization ratio of the feed, reducing the breeding cost, enhancing the disease resistance and production performance of the spotted deer, and realizing healthy and efficient breeding of the spotted deer.

A disclosed seed production method for cotton cytoplasmic male sterile line by utilizing insect hybridization is characterized by comprising: under the isolation condition, taking cotton cytoplasmic male sterilty male sterile line as a female parent, taking cotton restorer line as a male parent, planting the female parent and the male parent according a line number ratio of 1:4, and taking honeybee as pollination insect to perform cotton hybridization for seed production. According to the method, a box of honeybee can take the place of the pollination workload of one person, the seed production output at a normal year accounts for about 80% of the output of manual seed production, and thus labor is saved, labor intensity is reduced, seed production benefit is improved, and the purity and the quality of the hybridization seeds are guaranteed because of no selfing.

The invention discloses application of gRNA target combinations in construction of a hemophilia model pig cell line. The application comprises three gRNA target combinations, and specifically comprises the following steps: respectively designing a pair of target sequences aiming at pig F8 and F9 genes, constructing three CRISPR/Cas9 systems, respectively transferring the three CRISPR/ Cas9 systemsinto pig fibroblasts, and screening gene mutant strains, thereby obtaining the A, B and A & B hemophilia model pig cell line. According to the application disclosed by the invention, the gene editing efficiency is obviously improved by transforming the Cas9 expression vector. The editing efficiency is also obviously improved by adjusting the molar ratio of the gRNA expression vector to the Cas9 expression vector. The A, B and A & B hemophilia model pig can be obtained by applying a somatic cell nuclear transplantation cloning technology and is used for performing drug screening, drugefficacy evaluation, pharmacology and toxicology, disease pathology, gene therapy, cell therapy and other researches, effective experimental data is provided for further clinical application, and a powerful experimental means is provided for successfully treating human hemophilia.

The invention discloses a method for constructing high-quality porcine nuclear transplantation donor cells with high lean meat percentage, fast growth, high fertility and resistance to series epidemic diseases and application thereof. A CRISPR/Cas9 system for editing five genes of porcine MSTN, SST, INHA, CD163 and pAPN comprises a Cas9 expression vector and gRNA expression vectors respectively aiming at the porcine MSTN, SST, INHA, CD163 and pAPN genes; and the plasmid complete sequence of the Cas9 expression vector is shown as SEQ ID NO. 2. The corresponding gRNA expression vectors are respectively designed aiming at different target points of the MSTN, SST, INHA, CD163 and pAPN genes, and gRNA with higher editing efficiency and expression vectors thereof are obtained through screening. The gene editing is performed by matching with the modified Cas9 high-efficiency expression vector, and the editing efficiency is obviously improved compared with that of the original vector.

The invention discloses a pig feed for enhancing the pork flavor and a preparation method of the pig feed. The pig feed is prepared from the following raw materials: corns, soybeans, rice bran, fish and shrimp leftovers, fruit and vegetable leftovers, traditional Chinese medicines, edible salt, lycopene, bacillus natto, wine yeast, beer yeast, bifidobacterium, lactobacillus debrueckii and immune globulin. The pig feed for enhancing the pork flavor and the preparation method of the pig feed are reasonable in formula and high in palatability; soybeans are fermented by bacillus natto and then fermented with other grains through various microorganisms; by the adding of small-molecular nutritional substances, the pig feed is easy to digest; by the adding of various flavor substances, the flavor of pork is improved; the deliciousness, the fragrance and the sweetness are obviously enhanced, and the fishy smell is reduced; by the adding of the leftovers, wastes are turned into treasures, so that environmental pollution and treatment cost are reduced; the breeding cost is saved; by mixing of various traditional Chinese medicines, the disease resistance can be obviously enhanced, and the weight can be increased quickly; no additives are added, so that the pig feed is harmless to a human body and safe to eat.

The invention discloses a CRISPR/Cas9 system causing congenital deafness and application of the CRISPR/Cas9 system in preparation of model pig nuclear donor cells. The invention discloses a gRNA expression vector for a pig TMC1 gene. The gRNA expression vector expresses gRNA shown in SEQ ID NO: 25. The CRISPR/Cas9 system for editing the pig TMC1 gene comprises a Cas9 expression vector and the gRNAexpression vector disclosed by the invention, wherein the Cas9 expression vector is a plasmid with a complete sequence as shown in SEQ ID NO.2. Four gRNAs are designed for the pig TMC1 gene, efficient gRNAs are screened from the four gRNAs, and then preset targets are knocked out, so that the workload of later monoclonal cell identification and screening can be effectively reduced, and the gene editing efficiency can be detected directly through PCR product sequencing. When the Cas9 efficient expression vector modified by the system is used for gene editing, the editing efficiency is improvedby over 100 percent compared with that of the original vector.

The invention provides broiler feed using Cassia alata as the raw material and a production method of the broiler feed and relates to the technical field of feed. The broiler feed comprises corn flour, peanut cake, wheat, fermented bran, fish meal, bone meal, shell powder, silkworm chrysalis meal, the Cassia alata, Pennisetum purpureum Schum, waste tea leaf powder, salt, additives and an appropriate amount of water. The broiler feed has the advantages that the broiler feed does not generate any harmful substances from the formula to the production process of the broiler feed, broilers fed by the broiler feed are high in survival rate, high in resistance, and the meat of the broilers is tender, delicious, little in subcutaneous fat and rich in nutrition; in addition, all the raw materials of the broiler feed are combined to achieve synergic effects of promoting broiler growth and development, improving broiler meat quality and reducing broiler manure odor, and the broiler feed is efficient, safe, nontoxic and harmless.

The invention discloses a CRISPR/Cas9 system and application of the CRISPR/Cas9 system in construction of swine-derived recombinant cells for resisting amyotrophy protein gene defects. The invention provides an sgRNA combination, the sgRNA combination is composed of sgRNADMD-Ug3 (the target sequence binding region is shown as the 1-20th nucleotide in SEQ ID NO: 8) and sgRNADMD-Dg3 (the target sequence binding region is shown as the 1-20th nucleotide in SEQ ID NO: 11). The invention provides a kit, the kit is composed of a plasmid pKG-U6gRNA (DMD-Ug3) of sgRNADMD-Ug3 obtained through transcription, a plasmid pKGU6gRNA (DMD-Dg3) of sgRNADMD-Ug3 obtained through transcription, and a plasmid pKG-GE3. The sgRNA combination or the kit can be used for preparing recombinant cells or preparing a Duchenne muscular dystrophy animal model. The recombinant cells are anti-amyotrophy protein gene defect cells and can be used for preparing an animal disease model through somatic cell cloning. A solidfoundation is laid for the preparation of a Duchenne muscular dystrophy swine model, and has important application value for the research and development of Duchenne muscular dystrophy medicines.

The invention discloses a system for preparing severe immunodeficiency swine-derived recombinant cells with joint knockout of four RRIP genes, and particularly relates to a system for preparing severeimmunodeficiency swine-derived recombinant cells with joint knockout of four genes such as an RAG1 gene, an RAG2 gene, an IL2RG gene and a PRKDC gene. The invention provides an sgRNA (Ribonucleic Acid) combination. The sgRNA combination consists of sgRNARAG1-g4, sgRNARAG2-g2, sgRNAIL2RG-g7 and sgRNAAPRKDC-g6. The target sequence binding region of the sgRNARAG1-g4 is shown as nucleotides at the first to the twentieth site in SEQ ID NO: 9; the target sequence binding region of the sgRNARAG2-g2 is shown as nucleotides at the first to the twentieth site in SEQ ID NO: 13; the target sequence binding region of the sgRNAIL2RG-g7 is shown as nucleotides at the first to the twentieth site in SEQ ID NO: 24; and the target sequence binding region of the sgRNAPRKDC-g6 is shown as nucleotides at the first to the twentieth site in the SEQ ID NO: 34. The invention lays a solid foundation for preparation of a severe immunodeficiency swine model, and has important application values for research and development of severe immunodeficiency medicine.

The invention discloses a recombinant cell with IL2RG gene and ADA gene knocked out jointly and application of the recombinant cell in preparation of an immunodeficient swine model. The invention provides application of an sgRNA (small guide ribonucleic acid) combination consisting of sgRNAIL2RG-g7 and sgRNAADA-g7 in preparation of a kit. The invention further provides an sgRNA (small guide ribonucleic acid) combination. The sgRNA combination is composed of sgRNAIL2RG-g7 and sgRNAADA-g7. The target sequence binding region of the sgRNAIL2RG-g7 is shown as nucleotides at the first to the twentieth site in SEQ ID NO: 12; and the target sequence binding region of the sgRNAADA-g7 is shown as nucleotides at the first to the twentieth site in SEQ ID NO: 22. The invention also discloses application of the sgRNA combination in preparation of recombinant cells and application of the sgRNA combination in preparation of an immunodeficient animal model. The invention lays a solid foundation for preparation of a severe immunodeficiency swine model, and has important application values for research and development of severe immunodeficiency medicine.

The invention discloses a severe immunodeficiency swine-derived recombinant cell and a preparation method thereof. Specifically, the invention relates to a severe immunodeficiency swine-derived recombinant cell with joint knockout of three genes such as an IL2RG gene, an RAG1 gene and an RAG2 gene, a method for preparing the recombinant cell, and a kit for preparing the recombinant cell. The invention provides an sgRNA (small guide ribonucleic acid) combination. The sgRNA combination is composed of sgRNAIL2RG-g7, sgRNARAG1-g4 and sgRNARAG2-g2. The target sequence binding region of the sgRNAIL2RG-g7 is shown as nucleotides at the first to the twentieth site in SEQ ID NO: 24. The target sequence binding region of the sgRNARAG1-g4 is shown as nucleotides at the first to the twentieth site inSEQ ID NO: 9. The target sequence binding region of the sgRNARAG2-g2 is shown as nucleotides at the first to the twentieth site in SEQ ID NO: 13. The invention lays a solid foundation for preparationof a severe immunodeficiency swine model, and has important application values for research and development of severe immunodeficiency medicine.

The invention discloses a gene editing system for constructing high-quality porcine nuclear transplantation donor cells with high lean meat percentage and fast growth and application of the gene editing system. A CRISPR/Cas9 system for porcine SST-MSTN gene editing comprises a Cas9 expression vector, a gRNA expression vector aiming at a porcine MSTN gene and a gRNA expression vector aiming at a porcine SST gene. According to the invention, the corresponding gRNA expression vectors are designed aiming at different targets of the MSTN gene and the SST gene respectively, and gRNA with relatively high editing efficiency and the expression vector thereof are obtained through screening. The modified efficient Cas9 expression vector is used for gene editing cooperatively, and the editing efficiency is remarkably improved compared with that of an original vector.

The invention discloses a CRISPR system for ADCY3 gene editing and application of the CRISPR system to construction of obese porcine nuclear transplantation donor cells. The CRISPR/Cas9 system for porcine ADCY3 gene editing comprises a Cas9 expression vector and a gRNA expression vector for a porcine ADCY3 gene, wherein the Cas9 expression vector is a plasmid with a complete sequence as shown in SEQ ID NO.2, the gRNA expression vector expresses gRNA as shown in SEQ ID NO.22, and a target of the gRNA is as shown in SEQ ID NO.18. According to the invention, four gRNAs are designed for the porcine ADCY3 gene, efficient gRNAs are screened from the four gRNAs, then preset targets are knocked out, so that the workload of later monoclonal cell identification and screening can be effectively reduced, and the gene editing efficiency can be detected directly through PCR product sequencing.

The invention discloses a stocker with a large storage space for raising pigs. The stocker comprises a storage chamber, a through hole is formed in the storage chamber, a stirring bar penetrates through the through hole, an internal thread is arranged in the through hole, an external thread matched with the internal thread is arranged on the surface of the stirring bar, one end which protrudes outof the storage chamber of the stirring bar is connected with a reciprocating motor which drives the stirring bar to rotate around the axis of the stirring bar, two stirring blades are arranged at thelower end of the stirring bar, and a plurality of air holes are formed in each stirring blade; a feed inlet is formed in the upper end surface of the storage chamber, a cover plate is hinged to the side wall of the bottom end of the feed inlet, and a sealing gasket is adhered on the upper end surface of the cover plate; a discharge hole is formed in the lower end surface of the storage chamber, arubber plug is fixedly connected to the inner side wall of the discharge hole, and a feed opening is formed in the rubber plug; the moving and stirring in the length direction of the stirring bar oftwo stirring blades are achieved, thereby reducing the space of the storage chamber occupied by a stirring device.

The invention discloses a CRISPR system for constructing an MC4R gene mutated obesity porcine nuclear transfer donor cell and application of the CRISPR system. The system comprises a Cas9 expression vector and a gRNA expression vector for a porcine MC4R gene, wherein the Cas9 expression vector is a plasmid with a complete sequence as shown in SEQ ID NO.2, and the gRNA expression vector expresses gRNA as shown in SEQ ID NO.22; and the vector framework of the expression vector is pKG-U6gRNA, and the complete sequence of the plasmid is shown as SEQ ID NO.3. Gene editing is carried out by adopting the gRNA combined modified Cas9 high-efficiency expression vector screened by the invention, and the editing efficiency is remarkably improved compared with that of an original vector.