30 results about "Bovine muscle" patented technology

For stably and efficiently producing cloned livestock, used is a uniform cultured cell strain as a donor cell for nuclear transplantation.

Methods for in vitro isolation and culture and induced differentiation of bovine muscle satellite cells relate to the methods for in vitro isolation and culture and induced differentiation of cells. The invention solves the following problems: the cost is high and the heredity stability is not good in the existing method for isolation and culture of bovine muscle satellite cells, and the obtained myotubes are fewer and do not have contracting function in the existing method for induced differentiation of bovine muscle satellite cells. The method for in vitro isolation and culture of bovine muscle satellite cells is characterized by cleaning the tissues and digesting the tissues with trypsin and collagenase solution; carrying out centrifuging after filtering, re-suspending the cells and culturing the cells by a differential velocity adherent method after inoculation; and culturing the cells until the merging rate is 95% and carrying out heredity with pancreatin. The method for induced differentiation of bovine muscle satellite cells is characterized by obtaining PEI-plasmid complexes; culturing the bovine muscle satellite cells until the merging rate is 75%, adding high-glucose DMEM culture solution after cleaning, adding the PEI-plasmid complexes to the surfaces of the cells, changing culture solution A for culturing and then changing culture solution B for induced differentiation. In the method, the cost for isolation and culture of the bovine muscle satellite cells is low, the heredity stability is good and the myotubes obtained through induced differentiation are large in quantity and have contracting function.

The invention relates to a method for facilitating in vitro multiplication of a bovine skeletal muscle satellite cell. The method comprises the following steps: obtaining a target sequence containing a bovine miR-133b gene sequence; building an expression vector pEGFP-C1-miR133b; transfecting the bovine skeletal muscle satellite cell by the pEGFP-C1-miR133b. By application of the method for facilitating in vitro multiplication of the bovine skeletal muscle satellite cell, the in vitro multiplication ability of the bovine skeletal muscle satellite cell is improved by obtaining over-expression miniature RNA miR-133b of the bovine skeletal muscle satellite cell and using the effect of the miR-133b, and differentiation is effectively inhibited. The over-expression vector of the miR-133b is built by using the pEGFP-C1, and amplification and differentiation of a bovine muscle satellite cell are regulated by the miniature RNA, so as to obtain a high-purity bovine muscle satellite cell. A good cell model is provided for researching the function and the effect of each regulatory factor in amplification and differentiation processes of the muscle satellite cell.

The invention provides a method for knocking out a ZFNs (zinc finger nucleases)-mediated bovine MSTN (myostatin) gene and integrating an exogenous gene at the fixed point. The method comprises the following steps of: designing a specific locus expression vector of the ZFNs according to a bovine MSTN gene sequence; designing a target vector according to an acting site of the ZFNs, wherein the target vector contains the exogenous gene and can be integrated into a host genome; and transferring the expression vector and the target vector into a bovine fibroblast to obtain a cell for knocking out the bovine MSTN gene and integrating the exogenous gene at the fixed point. The DNA sequence acting on the designed specific ZFNs is located on the exon of the bovine MSTN gene. The constructed ZFNs-mediated target vector provides a way for easily and quickly knocking out the bovine MSTN gene and inserting the exogenous gene at the fixed point and has an important value to the genetic breeding of the new double-muscle bovine variety.

The invention belongs to the technical field of recombinant adenovirus vectors and discloses a method for constructing and packaging a bovine PDHB gene overexpression recombinant adenovirus vector. The sequence of the bovine PDHB gene overexpression recombinant adenovirus vector is GenBank: AY370909.2. Bovine PDHB gene CDS obtained through cloning is accordant with a sequence included in a database GenBank, the bovine PDHB gene CDS and a shuttle vector pAdTrack-CMV are recombined and are transfected with an HEK 293A cell, a recombinant adenovirus Ad-PDHB is successfully combined, and the titer of the recombinant adenovirus Ad-PDHB is 1.66*10<9>PFU.mL<1>; and after precursor fat cells in bovine muscles are infected by the adenovirus Ad-PDHB, the expression level of PDHB in mRNA is 25.5 times higher than that of a control group.

The invention provides a specific primer for detecting the mRNA expression levels of MSTN genes of cows. The specific primer is characterized by having the nucleotide sequence as follows: an upstream primer has the sequence: 5'-AACCAGGAGAAGATGGAC-3'; and a downstream primer has the sequence: 5'-TTAGAGGGTAACGACAGC-3'. The invention also provides a corresponding fluorescent quantitative PCR detecting kit. The specific primer can be used for achieving the aims of conveniently, rapidly and quantitatively detecting the mRNA transcriptional levels of MSTN genes, monitoring the muscle growth and development states of different cows (including milk cows, cattle and yaks), different tissues and different periods and also identifying diseases related to cow muscle dysplasia. The specific primer has the advantages of high sensitivity, good stability and low experimental cost on the aspect of detecting the transcriptional levels of the genes.

The invention discloses a Liupan mountain yellow cattle circR-UQCC1 gene, and an overexpression vector, a construction method and an application thereof, and belongs to the technical field of gene engineering. The invention discloses the annular Liupan mountain yellow cattle circR-UQCC1 gene for the first time; the recombinant overexpression vector capable of expressing the Liupan mountain yellowcattle circR-UQCC1 gene is constructed; after bovine muscle cells subjected to primary culture are transfected by the constructed vector, efficient expression of circR-UQCC1 mRNA can be obtained, anda foundation is laid for circR-UQCC1 gene function identification, cell transformation, metabolism regulation and growth and development.

The invention discloses a method for inhibiting proliferation and myogenic differentiation of bovine skeletal muscle satellite cells by interfering UBA2 expression, the method realizes inhibition of proliferation and myogenic differentiation of bovine skeletal muscle satellite cells by interfering UBA2 expression, and the genome base sequence of UBA2 is SEQ NO.1. According to the method, the proliferation and myogenic differentiation process of the bovine muscle satellite cells is regulated and controlled by changing UBA2 expression, enlightenment can be provided for research of ubiquitin-like modifier activating enzyme for muscle development and differentiation, and a new thought and reference are provided for clinical research and diagnosis and treatment of muscle development and differentiation and damage repair.

The invention discloses a method for enhancing estrus synchronization effects of cattle embryo transfer and used drugs, and specifically provides applications of luteinizing hormone-releasing hormoneA3 to preparation of drug compositions for enhancing the estrus synchronization effects of cattle embryo transfer. The method includes the following steps: providing donor cattle; injecting the drugsto the muscle of the donor cattle; continuously observing the estrus and ovulation conditions of cows in 3 days from the day that the drugs are injected, recording the beginning and ending time of estrus of the cows, performing observing in every morning and evening, determining estrus time, estrus procedure and ovulation status by combining the appearance estrus symptoms of the cows with rectal examination, determining estrus synchronization if the cows show estrus in 3 days after the drugs are injected, and timely performing artificial insemination on the cows ovulating in heat according totechnical operating instructions of artificial insemination of frozen semen, wherein semen is frozen semen from the same donor cattle; and after 90 d of mating, determining whether the cows are pregnant by taking rectal examination results as bases, and performing feeding during pregnancy according to conventional methods.

The invention belongs to the technical field of cell engineering, and particularly relates to an extraction method for separating muscle tissues of bovine muscle stem cells and application of the extraction method. The method comprises the following steps: (1) wiping the surface of cattle skin with alcohol, collecting muscular tissues of longissimus on the back of the cattle, and immediately rinsing with a phosphate buffer solution; (2) removing fascia and white film on the surface; and (3) rinsing with triple distilled water, and then rinsing with a phosphate buffer solution, alcohol and a phosphate buffer solution in sequence. The extraction method is different from the conventional method for treating by singly using a phosphate buffer solution, and the cost is reduced by combined treatment of three washing solutions. The method comprises the following steps: extracting muscle tissue, separating muscle stem cells from the extracted muscle tissue, using the muscle stem cells to construct a super enhancer, and optimizing and forming a method for analyzing the bovine muscle super enhancer by using an H3K4me1 and H3K27ac double-labeling method on the basis of chromosome co-immunoprecipitation high-throughput data basic analysis, which is different from a single labeling method in the prior art.

The invention discloses a method for detecting the expression level of an FASN gene in bovine muscle. According to the method, the expression level of the FASN gene in the bovine muscle is accuratelyand quickly analyzed through the design of an FASN gene-specific primer pair, the qPCR amplification of an internal reference primer GAPDH and relative quantitative analysis of a qPCR result. The qPCRdetection method for detecting the expression level of the FASN gene can be used for accurately detecting the expression level of the FASN genes in the muscle of cattle of different breeds. The method can be used for detecting the expression level of the FASN genes in the muscle of the cattle of different breeds, provides a reference for further research on the functions of the bovine FASN gene,and is simple and easy to operate.

The invention discloses a method for detecting HSL gene expression in bovine muscle. Through designing HSL gene specific primer pairs, amplification of internal primers of GAPDH qPCR, and the relativequantitative analysis to the result of the qPCR, the method for detecting HSL gene expression in the bovine muscle accurately and rapidly analysis the HSL gene expression in the bovine muscle. The established qPCR method for detecting HSL gene expression level can be used to accurately detect the HSL gene expression level of different cattle muscle. The method can be used to detect the expressionlevel of the bovine muscle HSL gene in different breeds, providing a reference for in-depth study of bovine HSL gene function. The method is easy and simple to operate.

The invention discloses a method for detecting the expression level of a PPAR gamma gene in bovine muscle. According to the method, the expression level of the PPAR gamma gene in the bovine muscle isaccurately and quickly analyzed through the design of a PPAR gamma gene-specific primer pair, the qPCR amplification of an internal reference primer GAPDH and relative quantitative analysis of a qPCRresult. The qPCR detection method for detecting the expression level of the PPAR gamma gene can be used for accurately detecting the expression level of the PPAR gamma genes in the muscle of cattle ofdifferent breeds. The method can be used for detecting the expression level of the PPAR gamma genes in the muscle of the cattle of different breeds, provides a reference for further research on the functions of the bovine PPAR gamma gene, and is simple and easy to operate.

The invention belongs to the technical field of gene engineering, and particularly relates to a bovine CFL2 gene adenovirus interference vector and a construction and identification method thereof.The bovine CFL2 gene adenovirus interference vector is characterized in that Ad-CFL2 shRNA-1 with a good interference effect is used for infecting bovine primary muscle cells, packaging and propagation of adenovirus are carried out, and by constructing the bovine CFL2 gene adenovirus interference vector, the bovine CFL2 gene adenovirus interference vector can be obtained. The purpose of continuously knocking down CFL2 gene expression in bovine muscle tissues and cells can be achieved, an effective technical support is provided for follow-up research on a molecular regulation mechanism between the CFL2 gene and muscle growth and development, and a new molecular marker and breeding thought can be further provided for genetic improvement and efficient breeding of local cattle in China.

A protein mixture of meat extract and collagen from bovine muscle takes the form of a soluble powder composed of macronutrients such as carbohydrates, lipids and proteins, micronutrients such as calcium, iron and sodium, and saturated fat. It is therefore a balanced formulation that preserves all essential amino acids, represents a source of iron and calcium and exhibits a low sodium and saturated fat content. The protein mixture can be used with any type of edible liquid or paste, in the form of food supplements, enteral or parenteral fortifying diets, and is particularly recommended for patients or persons suffering from some form of deglutition disorder.

The invention provides a construction method of an oxidative damage model for a muscle cell of a beef cattle and the application of the oxidative damage model, and belongs to the technical field of cytobiology. The invention adopts the technical scheme that the construction method of the oxidative damage model for the muscle cell of the beef cattle comprises the following steps: preparing a cell culture fluid, separating a beef cattle muscle tissue, culturing the muscle cell of the beef cattle, and acquiring the construction parameter of the oxidative damage model. The construction method hasthe beneficial effects that the separated muscle cell of the beef cattle is taken as the object of study, the oxidative damage model for the muscle cell of the beef cattle is established through hydrogen peroxide, the materials are provided for studying the reduction of the beef quality and studying the nutrition control mechanism, the research and development costs for the subsequent tests are reduced; the established oxidative damage model for the muscle cell of the beef cattle provides the mechanism studying materials for studying the control mechanism of the beef quality and developing anantioxidant active material, and the time and the cost for researches and development are greatly shortened and reduced by directly applying the model.

The invention relates to a bovine lncRNA-133a sequence and its application and verification method in the regulation of proliferation and differentiation of bovine skeletal muscle satellite cells. The present invention provides a sequence of long-chain non-coding RNA lncRNA-133a related to bovine muscle development. The present invention finds for the first time that lncRNA-133a has the function of promoting the proliferation and myogenic differentiation of bovine skeletal muscle satellite cells, and can apply lncRNA-133a In regulation of proliferation and differentiation of bovine skeletal muscle satellite cells.

The invention discloses a method for increasing the availability of receptor cattle in bovine embryo transplantation. By injecting luteinizing hormone-releasing hormone A3 into cows, the technical effect of one or more aspects of the invention can be obtained. The method comprises the following steps: (1) providing a donor cattle; (2) intramuscularly injecting luteinizing hormone-releasing hormoneA3 into the donor cattle; (3) continuously observing oestrus and ovulation conditions of the cow within 3 days from medicine injection, recording oestrus beginning time and oestrus ending time of thecow, observing the oestrus beginning time and the oestrus ending time once every morning and evening, determining oestrus time, oestrus procedures and ovulation conditions by combining rectal examination according to the oestrus symptoms of the cow, judging estrus within 3 days after medicine injection as estrus synchronization, and performing artificial insemination on the estrus ovulated cows at proper time according to operating rules of a freezing technology, with semen being frozen semen of the cows of the same donor; and (4) determining whether the cow is pregnant or not according to arectal examination result 90d after hybridization, and feeding the cow during pregnancy according to a conventional method.

The invention relates to a method for promoting myogenic differentiation of bovine skeletal muscle satellite cells by overexpression of PSMB5. The method comprises the following steps: (1) obtaining atarget sequence containing a bovine PSMB5 gene sequence; (2) constructing an expression vector pcDNA3.1-PSMB5; (3) transfecting bovine skeletal muscle satellite cells by pcDNA 3.1-PSMB5, using the obtained expression vector pcDNA3.1-PSMB5 for transfecting bovine skeletal muscle satellite cells by adopting a liposome transfection reagent so that the bovine skeletal muscle satellite cells overexpress the PSMB5, and improving the in-vitro differentiation capacity of the bovine skeletal muscle satellite cells by utilizing the effect of the PSMB5. According to the method, PSMB5 is used for regulating and controlling proliferation and myogenic differentiation of bovine muscle satellite cells, the enlightenment can be provided for research on the ubiquitination process of muscle development anddifferentiation, and a new thought and reference are provided for clinical research and diagnosis and treatment of muscle development and differentiation and injury repair.

The invention belongs to the field of cell culture, and discloses a bovine muscle stem cell in-vitro separation culture method, a culture medium and application thereof. According to the muscle stem cell in-vitro culture method, muscle stem cells are subjected to in-vitro culture by adopting a cell culture medium of non-animal-derived hydrolysate. The invention further discloses a culture medium for culturing the muscle stem cells. By adopting the culture medium disclosed by the invention, the muscle stem cells with ideal cell quantity and cell morphology can be obtained on the basis of not adding or adding a small amount of serum, and the muscle stem cells can still maintain dryness and differentiation potential after continuous passage for multiple times. The muscle stem cells cultured in vitro are used in the field of cell culture meat, and the method has a wide application prospect.