Cow lysozyme (Lyz) gene mammary gland specific expression recombinant plasmid as well as construction method and application thereof

Abstract

The invention provides a cow lysozyme (Lyz) gene mammary gland specific expression plasmid as well as a construction method and application thereof. Specifically, a coding sequence of a Lyz gene is subjected to PCR amplification, the Lyz gene is inserted into a multiple cloning site of a universal expression vector pEGFP-N1 carrying an enhanced green fluorescent protein gene; Ase I+Hind III doubleenzyme digestion is conducted on the universal expression vector pEGFP-N1 so as to remove a CMV promoter, a cow mammary gland specific expression beta-lactoglobulin gene (BLG) promoter sequence is cloned, Ase I and HindIII restriction enzyme cutting sites are replaced, and the cow Lyz gene mammary gland specific expression plasmid is constructed. The recombinant plasmid is adopted to transfect cow mammary epithelial cells, and an obtained cell line can be used for secreting cow lysozyme with biological activity and used for cow mammary tissue immune mechanism research; and the recombinant plasmid can also be applied to research and development of preparations for gene therapy of cow mastitis.

Description

technical field [0001] The invention relates to a mammary gland-specific expression recombinant plasmid of cow lysozyme gene and its construction method and application, belonging to the technical field of gene recombination and molecular cloning. Background technique [0002] Dairy cow mastitis is a common and frequently-occurring disease in dairy cows. As the incidence of dairy cow mastitis increases, the milk production of dairy cows will be greatly reduced, and the quality of raw milk will be seriously affected. Therefore, the treatment of cow mastitis is getting more and more attention. At present, the main treatment methods are antibiotics and traditional Chinese medicine preparations. The use of antibiotics, the generation of drug-resistant strains and drug residues in milk seriously endanger the health of consumers. Therefore, the treatment of cow mastitis has become a difficult problem in the world. Use less or no antibiotics, effectively control drug residues, and...

AI Technical Summary

Problems solved by technology

At present, the main treatment methods are antibiotics and traditional Chinese medicine preparations. However, the use of antibiotics, the emergence of drug-resistant strains and drug residues in milk seriously endanger the health of consumers. Therefore, the treatment of cow mastitis has become a difficult problem in the world.

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