42 results about "Somatic Cell Nuclear Transfer Technique" patented technology

The invention provides a method of cloning a reproductive and respiratory syndrome resisting pig. The method comprises the following steps: transferring CRISPR / Cas9 targeting vectors and CD163 gene homologous recombination modification vectors into fibroblasts of a pig to obtain positive clone cells, wherein the seventh exon of the endogenous CD163 gene in the positive clone cells of the pig is replaced by the tenth exon of the CD163-L1 gene of human, so that the positive clone cells are incapable of mediating PRRSV invading; obtaining a clone embryo by adopting a somatic cell nucleus transfer technology by taking positive cells as nucleus transfer donor cells and oocytes as nucleus transfer recipient cells; and transferring the clone embryo into the uterus of the pig, and impregnating to obtain a cloned pig. The method provided by the invention is low in cost, the time of obtaining the homozygous pig is shortened greatly, the expression of the CD163 gene is not affected by gene editing, and a foundation is laid for the research into the gene function of large animals and the establishment of disease models on the basis of CRISPR / Cas9 mediated homologous recombination.

The invention discloses a transgenic vector of salivary gland tissue specific expression foreign protein and a construction method thereof; the expression vector is constructed by inserting a recombinant fusion gene constructed by glucanase gene, xylanase gene, and phytase gene into a parotid gland-specific transgenic vector; the vector is introduced into pig genome, and transgenic pigs is produced by a somatic cell nucleus transplant technique. The transgenic vector of the invention has hydrolytic action on glucanase in barley raw materials, xylanase in wheat and corn, and phytase in vegetable feed; the pig salivary gland is used as a reactor to realize secretion for the whole life; the addition of the enzyme preparations into feed is replaced permanently, which gives better play to the enzymes; the expression vector of the invention has a piggyback transposon, which greatly increases the transgenosis efficiency, changes the transgenic integration model of common plasmid serial recombination into single locus single copy integration, and thus better simulates the internal environment of biological genes.

The invention relates to a reconstructed ovum of an albinism model pig and a construction method thereof, and a construction method of the model pig. According to the construction method of the reconstructed ovum, tyrosinase is knocked out to obtain the reconstructed ovum of the albinism model pig. The combination of a CRISPP / Cas9 gene knock-out technique and a somatic nucleus transplantation technique proves that the method for constructing the gene modified pig has high feasibility. The obtained TYR gene knock-out pig has the typical characteristics of albinism, can provide a reliable animal model for researching human albinism, can be applied to toxicity and anaphylaxis detection and many other aspects, and is hopeful to become a standardized experimental animal for albinism-like mice.