41 results about "Porcine muscle" patented technology

The invention relates to a separation and purification method of high-purity porcine muscle stem cells, for research and production of cultured meat. The separation and purification method disclosed by the invention comprises the steps of obtaining sterile muscular tissue, efficiently digesting the muscular tissue to obtain a large quantity of muscle mononuclear cells, and finally performing steaming cell sorting to obtain the high-purity porcine muscle stem cells. The invention further discloses a method for efficiently digesting the porcine muscular tissue. By the method, the digestion speedand the digestion intensity can be increased, and a large quantity of muscle mononuclear cells can be obtained. When the purification method disclosed by the invention is applied, the porcine musclestem cells of which the purity exceeds 92% (by PAX7) can be stably obtained. The porcine muscle stem cells obtained through separation have efficient disintegration efficiency, and MYOSIN expression quantity can reach 85%-90%. When the method disclosed by the invention is applied, a large quantity of seed cell sources can be provided for research and actual production of the cultured meat.

The invention belongs to the technical field of preparation of a pig genetic marker, and particularly relates to a preparation method of a pig muscle pH value related molecular marker and application of the pig muscle pH value related molecular marker on auxiliary marker selection. The molecular marker is obtained by cloning a GYS1 gene promoter sequence, and the nucleotide sequence of the molecular marker is as shown in the sequence table SEQ ID NO: 1; and a C267-T267 basic group on the 267th basic group of a sequence as shown in the sequence table SEQ ID NO: 2 is mutated to cause MvaIPCR-RFLP polymorphism. The invention also discloses application of the preparation method for obtaining the molecular marker and a polymorphism detection method. The preparation method disclosed by the invention provides a new genetic marker for carrying out the auxiliary marker selection of a high-muscle quality pig.

The invention belongs to the technical field of preparation of livestock molecular markers, and particularly relates to an SNP (Single Nucleotide Polymorphism) molecular marker related to a pH (Potential of Hydrogen) value character of pig muscle. The molecular marker is obtained by LDHA (Lactate DehydrogenaseHydroxyacyl) gene cloning and has a nucleotide sequence as shown in Figure 4.In the sequence as shown in Figure 4, the 495th position has allele G or A base substitution, and the 635th position of the sequence has allele C or T base substitution. The new molecular marker is provided for assistant selection of pig markers.

The invention discloses an lncRNA marker related to porcine muscle fiber types and an application of the lncRNA marker, and belongs to the technical field of livestock molecular marker preparation. The lncRNA marker is LincRNA-ROMT, and the sequence of the lncRNA marker is as shown in SEQ ID NO:1. The LincRNA-ROMT is closely related to the porcine muscle fiber types, and extremely remarkably showslow expression in oxidized porcine muscle fibers (fat type Lantang porcine muscle tissues). The lncRNA marker can be applied to an auxiliary genetic breeding process in the aspect of pig variety improvement. The LincRNA-ROMT disclosed by the invention can be used as a molecular marker related to porcine muscle fiber types, and has important practical application values in genetic assisted breeding.

The invention discloses a method for detecting the content of triglyceride in porcine muscle with gene PPARG. The expression level of gene PPARG in adipose tissue is utilized to detect the content of triglyceride in porcine muscle. The method has the characteristics of rapidity, accuracy, convenient operation and no influence to the normal growth of pigs, is suitable for dynamic monitoring of the content of triglyceride in porcine muscle during pig breeding so as to grasp the situation of pig growth at any time, can achieve relatively accurate understanding of the content of fat in porcine muscle, and then adjusts the daily ration of pigs.

The invention discloses a construction method of a porcine FLJ small interference RNA expression vector. The construction method comprises the following steps of: 1) designing 3 pairs of siRNA primers of loop rings containing 9 nucleotides, wherein the 9 nucleotides are TTCAAGAGA; and 2) constructing an RNAi expression vector, namely after annealing the siRNA primers, connecting the siRNA primers with double-enzyme-cut RNAi vector and converting to obtain recombinant plasmid of the RNAi expression vector. The invention also provides a method for identifying the constructed porcine FLJ effective small interference RNA expression vector. The invention further provides application of the porcine FLJ effective small interference RNA expression vector constructed by the method. The porcine FLJ effective small interference RNA expression vector is used for researching the influence of the FLJ gene on fat deposition in porcine muscle.

The invention discloses a long non-coding RNA marker Lnc-482286 related to porcine skeletal muscle satellite cell proliferation, and application of the long non-coding RNA marker Lnc-482286, and belongs to the technical field of molecular biology. The long non-coding RNA marker is Lnc-482286, and the nucleotide sequence of the long non-coding RNA marker Lnc-482286 is shown as SEQ ID NO.1. The marker Lnc-482286 provided by the invention is highly expressed in porcine muscle tissues, expression of the Lnc-482286 in the muscle tissue of the Jinfen white pig is proved to be in a trend of firstly increasing and then decreasing along with the increase of the day age, and the Lnc-482286 plays a significant up-regulation role in the process of the porcine skeletal muscle satellite cell proliferation. The proliferation condition of the porcine skeletal muscle satellite cells can be identified by detecting the expression of the Lnc-482286, the porcine skeletal muscle satellite cell proliferation can be accelerated by improving the expression of the Lnc-482286, and a new thought and method is provided for clinical research on the growth and development of the porcine skeletal muscle.

The invention discloses a method for low-cost in-vitro culture of porcine muscle stem cells, and belongs to the technical field of stem cell and animal cell culture. The invention discloses a culturemedium and the culture method for culturing the muscle stem cells at low cost, the muscle stem cells can keep in-vitro growth and myogenic differentiation capacity through the culture medium and the culture method, and the cost of the culture medium is greatly reduced.

The invention specifically relates to construction and application of over-expression vectors of three different subtypes of porcine PDLIM3 protein, belonging to the field of bioengineering technology. The nucleotide sequences of the gene coding regions of the three different subtypes of the porcine PDLIM3 protein are shown in SEQ ID No. 1, SEQ ID No. 2 and SEQ ID No. 3 in a sequence table. Three over-expression vectors of the three subtypes of the porcine PDLIM3 protein can be simultaneously constructed by subjecting the complete coding region fragments of the genes of the three subtypes of the porcine PDLIM3 protein to double enzyme restriction and connecting the treated fragments with the pEGFP-N1 vector having undergone double enzyme restriction. The above-mentioned eukaryotic expression vector construction method is simple and practicable; the over-expression vectors can highly-efficiently enhance expression of the PDLIM3 protein, all contain green fluorescent protein and can be used for detecting expression and location of the three different subtypes of the porcine PDLIM3 protein in eukaryotic cells. The over-expression vectors provided by the invention are also applicable to research on roles of the porcine PDLIM3 protein in growth and development of porcine muscle fibers.

The invention discloses an efficient and economical pig muscle stem cell separation method and application thereof, and belongs to the technical field of biology. According to the method, different enzymes for digesting muscle tissues are combined, the single cell releasing capacity of different combined enzymes is determined, the most efficient and economical enzyme combined digestion mode (the combination of streptavidin and dispersozyme) is selected, and finally a large number of muscle stem cells are obtained through a differential purification method. The invention further discloses a method for efficiently digesting the pig muscle tissue, and the muscle stem cells can be maximally added while the cost is reduced. By applying the enzyme combined digestion and separation method, the total single cell number of 4.6-6 * 10 < 6 > cells/g and the pig muscle stem cell number of 8-15 * 10 < 5 > cells/g (in a double positive proportion of CD29 and CD56) can be stably obtained. The muscle stem cells obtained through separation have high differentiation capacity, and an efficient and economical separation method is provided for research and actual production of cell culture meat seed cells.

The invention discloses a molecular marker related to pork quality characters and application of the molecular marker, and belongs to the field of molecular markers. The nucleotide of the molecular marker is shown as SEQ ID NO: 1, C/T mutation exists at the 302bp position of the sequence shown as SEQ ID NO: 1, the mutation comprises three mutation types of CC, CT and TT, and when the genotype of a pig individual is CC type, the contents of marbling of longissimus dorsi muscle, marbling of biceps femoris muscle and intramuscular fat of the longissimus dorsi muscle of the pig individual are higher than those of a CT type pig individual and a TT type pig individual. The invention further provides a primer pair for detecting the site, a corresponding detection method is established, and a new molecular marker is provided for genetic improvement of pig muscle quality. According to the present invention, the pork quality character can be subjected to early stage selection, and the detection method is efficient and accurate, such that the pork quality improvement process can be accelerated, and the economic benefits of pig breeding can be improved.

The invention discloses a method for identifying diameters of muscle fibers of a pig and a primer pair used by the method. The primer pair amplifies a DNA fragment containing a polymorphism site whereSSC4 g. 69991162A is larger than G; the polymorphism site where the SSC4 g. 69991162A is larger than the G is the 69991162 nucleotide from the 5' terminal on the 4 chromosome of a porcine reference genome Sscrofa11.1; the porcine reference genome Sscrofa11.1 is a porcine reference genome sequence in GenBank, wherein the updated date is February, 2017. The method can perform early screening on to-be-selected pigs, so that the problem of long time for selecting good breeding pigs in actual production is effectively solved, the breeding cost is reduced, the diameters of the porcine muscle fibers are effectively reduced or increased, the method has high accuracy and low detection cost, automatic detection can also be achieved, and the practical application value in pig breeding is very high.

Embodiments of the invention disclose an SNP molecular marker associated with drip loss of muscles of a Yorkshire pig and application of the SNP molecular marker. The SNP molecular marker is the polymorphic nucleotide site rs81470247 of an intron variant of the pig gene HOXA5; and the nucleotide sequences of upstream and downstream 100 bp of the site are as shown in SEQ ID No. 1, the 101 base of the sequence is mutated, and the base K is T or G, resulting in polymorphism. The invention further develops a primer for the SNP molecular marker. A method for screening a strain of a Yorkshire pig with low drip loss by identifying the SNP marker is established on the above basis, so breeding of the strain of the Yorkshire pig with low drip loss, breeding of high-quality pork or content detection is aided, and the pork has important edible value, economic benefits and social value.

The invention discloses an antibiotic-free crude feed suitable for free-range black pigs and an application of the antibiotic-free crude feed. The antibiotic-free crude feed is characterized by comprising the ration formula components and content of porcine muscle fat deposition after functional conversion. The antibiotic-free crude feed prepared by the method comprises the ration formula components and nutrient value, on one hand, the quality of meat of live pigs can be notably improved, and on the other hand, the nutrient transformation efficiency of the feed of variety pigs can be improved,and the discharge amount of nitrogen, phosphorus, copper and zinc in excrement can be greatly reduced.